UGC Sponsored

National Conference on

Recent Advancements in
Science and Technology

VOLUME I : Chemistry

:: Organized by ::

Vidya Bharati Shaikshanik Mandal's

Vidya Bharati Mahavidyalaya, Amravati

Re-accredited with Grade 'A' by the NAAC (CGPA 3.23 - Third Cycle)
College with Potential for Excellence (CPE Status Thrice by the UGC)
Star College Status by DBT, New Delhi, Mentor College under Paramarsh by UGC
Identified as Lead College by S.G.B. Amravati University, Amravati

:: In Collaboration with ::

S.S.S.K.R. Innani Mahavidyalaya

Karanja (Lad), Dist. Washim 444105 (M.S.), India
NAAC Re-Accredoted 'A+' Grade (CGPA 3.28)
C.P.E. Status Awarded by UGC, New Delhi
Affiliated to Sant Gadge Baba Amravati University, Amravati

ISBN : 978-81-19931-25-5
 UGC Sponsored
National Conference on

Recent Advancements in
Science and Technology

VOLUME I : Chemistry

:: Organized by ::

Vidya Bharati Shaikshanik Mandal's

Vidya Bharati Mahavidyalaya, Amravati

Re-accredited with Grade 'A' by the NAAC (CGPA 3.23 - Third Cycle)
College with Potential for Excellence (CPE Status Thrice by the UGC)
Star College Status by DBT, New Delhi, Mentor College under Paramarsh by UGC
Identified as Lead College by S.G.B. Amravati University, Amravati

:: In Collaboration with ::

S.S.S.K.R. Innani Mahavidyalaya

Karanja (Lad), Dist. Washim 444105 (M.S.), India
NAAC Re-Accredoted 'A+' Grade (CGPA 3.28)
C.P.E. Status Awarded by UGC, New Delhi
Affiliated to Sant Gadge Baba Amravati University, Amravati

SAI JYOTI PUBLICATION, NAGPUR

UGC Sponsored
National Conference on
=====================================
Recent Advancements in Science and Technology
=====================================
VOLUME I : Chemistry
=====================================

Chief Editor
Dr. Pradnya S. Yenkar
Principal, Vidya Bharati Mahavidyalaya, Amravati

ISBN : 978-81-19931-25-5
th
Date : 10 Feb., 2024

Publisher :
Sai Jyoti Publication
Itwari, Nagpur
E-mail : [email protected]

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LASER POINT,
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No part of this book shall be reproduced, stored in retrieval system, or translated in

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Disclaimer :
The publishers, The Principal, Vidyabharti Mahavidyalaya, Amravati and
the organizers are not responsible for the content of any of the research
paper/article published in this conference proceedings.
The authors/Co-authors are responsible for the plagiarism
For any conflict of interests, the Author/Co-Authors are responsible.
For any query/issue/ethical aspects contact the corresponding Author.
 CONFERENCE PROCEEDINGS
UGC Sponsored National Conference on
Recent Advancements in Science & Technology

Date : 10th Feb, 2024

:: Organized by ::
Vidya Bharati Shaikshanik Mandal's
Vidya Bharati Mahavidyalaya, Amravati
:: In Collaboration with ::
S.S.S.K.R. Innani Mahavidyalaya
Karanja (Lad), Dist. Washim 444105 (M.S.), India

Editorial Board

VOLUME I : Chemistry

Chief Editor

Dr. Pradnya S. Yenkar

Principal,
Vidya Bharati Mahavidyalaya, Amravati

Editors

Prof. Dr. M. M. Rathore, Associate Editor

Dr. V. H. Masand, Member
Dr. P. S. Bodkhe, Member
Dr. C. N. Deshmukh, Member
Mr. Varun Mahale, Member
Dr. P. P. Nalawade, Member
Dr. J. R. Bansod, Member
Mr. C. N. Jadhav, Member
.
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

CONTENTS
Chemistry
S.No. Title Author/s Page No.
1 QSAR evaluation of Glucagon Ajaykumar Gandhi 1-7
Receptor (GCGR) antagonists for Pooja Gaikwad
the anti-hyperglycaemic lead Jyoti Dahatonde
development Archana Chapolikar
2 Synthesis and Bilogical Evaluation A P Thakare 8-12
of Pyrazole Derivative S R Kolteke
3 Healthy hydroponic cattle feed Dr. Ramesh Tukaram 13-18
(grass) development eco-friendly & Parihar,
economically Dr A.G.Gaddamwar
4 Design and Synthesis of Somwanshi A.R. 19-22
Heterocyclic Active Moieties of
Pyrazoles and Its Importance’s
5 QSAR analysis of sodium glucose Archana Chapolikar, 23-34
co–transporter 1 (SGLT1) inhibitors Jyoti Dahatonde,
for anti-hyperglycaemic lead Pooja Gaikwad,
development Ajaykumar Gandhi
6 Preparation and Acetone Gas A. V. Kadu 35-40
Sensing Properties of Cd Substituted
Magnesium Ferrites
7 Studies of Acoustic Properties of A. D. Khambre 41-44
Substituted Chalcone in Different
Percentage of Dioxane-Water
Mixture
8 Synthesis, Physicochemical Girish Deshmukh 45-49
evaluation of heteryl amino Avinash Thakare,
derivatives of bis[5-cyano-1,6- Chanda Gawande
dihydro-6-imino-2-isopropyl-4-
(methylthio) pyrimidine] diazene
9 Novel Synthesis and Chhaya D. Badnakhe 50-58
Characterization of Some
Substituted Chlorosubstituted Aryl
Substituted 1,3-Thiazole as
Antibacterial Agents
10 Mini Review on biological aspects Mr. N.D. Dahake, 59-61
of 1,3,4-Thiadiazoles Dr. V.H. Masand
11 Simultaneous Non-aqueous Pradip P. Deohate, 62-64
Conductometric Investigation of Aakanksha S. Dongare
Pharmaceutically Potent Ibuprofen- Anushri R. Jalamkar
Paracetamol Combination Drugs
12 Cp*Co(III) Catalyzed Redox- Dewal S. Deshmukh 65-67
Neutral Annulation: A Powerful Bhalchandra M. Bhanage
Tool for Isoquinoline Synthesis
from N-Cbz Hydrazones
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

13 Design, synthesis, spectral K. U. Dongare 68-72

characterization and anti-microbial S. A. Waghmare
screening of some novel 1, 2, 4- S. B. Sarkate
dithiazole with Coumarin moiety R. N. Ingole
14 Effect Of metal-ligand complexon Prof. M.M.Rathore 73-78
germination of some vegetable C.N.Deshmukh
plants
15 Preparation of Adsorbent from Sandhya B. Gaikwad 79-82
various natural materials for
removal of heavy metal ions from
waste water: Critical review
16 Evaluation of Antioxidant Activity P.L. Harale 83-86
by DPPH Method of M.E. Shelke
Thiocarbamides D.T. Tayade
17 Investigations of Proximate Jawerea Nayab Mohammad 87-89
Compositions of Xanthium Jamil
Strumarium L Roots from Tisgaon Rahim Ullah Sharafat Ullah
(PIN Code 431002) of Taluka and
District Aurangabad, India
18 Synthesis, Structural Study Of Kavita. M. Heda 90-93
Substituted Heterocyclic
Compounds
19 Characterization of Diesel Mixtures K. P. Belsare 94-96
by Ultrasonic Methods N. B. Selukar
20 Phytochemical and Proximate Miss Mosim Raza H. Shaikh 97-100
Analysis of Leaves of Heteropogon
Contortous (L.) Beauv
21 A Review of Computer-Aided Drug Manojkumar O. Malpani 101-105
Discovery (CADD)
22 Synthesis and Anti-microbial M. R. Raghuvanshi 106-109
Activity of Novel 3-[(3-
substitutedamino-1,2,4-thiadiaz-3-
yl)]amino-5-N-TAG-amino-1,2,4-
thiadiazole [11b]
23 Sulfate promoted zirconia-based Sushil V. Shelke 110-112
mixed oxide as solid acid catalysts Sambhaji T. Dhumal
for organic transformations Abhay L. Ghodke
Meghshyam K. Patil
24 Structure, Physical and Chemical A.P. Mitake 113-119
Properties of Thiourea with Their S.P. Rathod
Derivatives
25 Study of Proton and Metal-Ligand Mohd. Wajid Shaikh Mohd 120-123
Stability Constants of hydroxy Waris
Substituted Chalcone Complexes
pH-Metrically
26 Ecofriendly synthesis and Nilesh B. Jadhav 124-126
characterization of Ethyl 2-imino-6-
methyl-4-substituted phenyl-1, 2, 3,
4-tetrahydropyrimidine-5-
carboxylate compounds
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

27 Effect of Fe Dopant on Structural & N. N. Gour 127

Electrical Properties of Copper A. P. Pinjarkar
OxideNanoparticles H. G. Wankhade
P. R. Padole
28 Preliminary Phytochemical Studies Nitin G. Asole 128-131
of Cesalpinia crista L. Ramesh T. Parihar
Anand S. Jadhao
29 Removal of Methylene Blue using Vilayatkar N.D. 132-134
Cajanus Cajan Activated Carbon Chaudhari D.L.
Kadu N.S.
Kurzekar R.R.
30 Conductometric Studies of K.P. Jumde 135-140
[CPHDD] and [CTMBCD] at Sanghapal S. Padhen
Different Molar Concentrations and Saleem R. Khan
Different Percentage Compositions D.T.Tayade
in Water-Ethanol Mixture
31 Synthesis and Characterization of Nilesh S. Padole 141-144
Some Substituted Chalcone by the Murlidhar P. Wadekar
Green Synthesis Way (Grinding Shubhangi Y. Deshmukh
Method) Vinod M. Sherekar
32 In-Silico Prediction of Pooja.P.Patle 145-156
Phytoconstituents From Parimal Katolkar
Solanum Indicum for Antiepileptic Pradeep Raghatate
Activity Jagdish Baheti
33 Deep Discoveries in Chemistry: Dr. Prashant R. Mahalle 157-159
Unraveling the Mysteries at the
Molecular Level
34 Synthesis, Characterization of (3- Ghodile R.D 160-164
Chloro-benzylidene) – (4-Phenyl- Chaudhari P.S.
thiazol-2-yl) amine and Studied their Dharamkar R. R
antioxidant Activity
35 Synthesis and Characterization of Priyadarshani N Deshmukh 165-167
Schiff Bases Using Different
Amines and their Biological
Activities
36 Physico-Chemical Assessment of Dr. Priyanka U. Belsare 168-171
Soil Sample in Jarud Region in Ku Tejashri M. Ghotkar
Amravati District of Maharashtra
(India)
37 Synthesis ,Characterization And P. M. Dahikar 172-175
Antimicrobial Study Of
Co(II),Cu(II) and Fe(III) Complexes
Of Substituted 4,4'-
Dimethoxybenzoinhydrazones
38 Study of Extraction and Chemical Ramesh Tukaram Parihar 176-178
Screening of T. cordifolia (Guduchi) S A Quazi
Stem & Leaves Shaikh Farah T
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

39 In-silico QSAR-based virtual Rahul D. Jawarkar 179-196

screening recognition of novel Praveen Sharma
anaphylactic lymphoma kinase Sachin Kumar Jain
inhibitor
40 Synthesis, characterization and Rajendra M. Pathade 197-203
biocidal study of some novel 2- Pravin. S. Bodkhe
hydroxy 4,6-disubstituted phenyl
pyrimidines bearing 4-bromo phenol
moiety
41 Investigation of antimicrobial Manish Fukate 204-208
Activities of Dihydropyrazines Sheryil Malvilayils
Roshan Jaiswal
Swati Bawiskar
42 Synthesis and Spectral Mr Ramdas N Ingole 209-211
Characterization of Novel 1,2,4- Dr Siddharth S Waghmare
Dithiazole Derivatives Dr Pravin S Bodkhe
43 Synthesis of Substituted Flavanones S. P. Rathod 212-215
and their Impact on Seed R.T.Parihar
Germinations A.P.Mitake
T.M.Bhagat
S. B. Waghmare
44 Antimicrobial Resistance : A Global Reshal Deshmukh 216-217
Challenge
45 Wings across Reservoirs: Dr. Chandrashekhar R. Kasar 218-222
Understanding and Safeguarding the Ku. Revati Kishor Lonkar
Migration Patterns of Insectivorous
Birds in Anthropogenic Landscapes
46 Synthesis and Characterization of Mr. R. N. Gaikwad 223-225
Pyrazolone and its derivatives
47 Preliminary Phytochemical analysis Swapnil D. Bhagat 226-229
of Leaves extract of the plant Sopan D. Ingole
Justicia Varun A. Mahale
Adhatoda Nandkishor S. Thakare
Chandrakant U. Dhanwad
48 In-Silico Prediction of Sakshi Mude 230-241
Phytoconstituents From Parimala Katolkar
Solanum Indicum for Antistress Pradeep Raghatate
Activity Targeting Ask 1 Inhibitor Jagdish Baheti
49 Design, Synthesis, Spectral S. B. Sarkatea 242-245
Characterisation and Antibacterial S. A. Waghmare
Screening of Some Novel 4- K. U. Dongare
Substitutedimino-1,3,5-Dithiazine R. N. Ingole
Along With Pyrimidine Nucleus

50 Comparative Account of Partial R. V. Dudhate 246-249

Molar Volumes and H. N. Pawar
Compressibilities of Aqueous-(L- B. R. Bhosle
Arginine + Glucose/Lactose) S. D. Deosarkar
Solutions at 310.15 K S. D. Deshmukh
P. S. Bodkhe
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

51 Studies in the Synthesis of 5-(3- M. B. Shahakar 250-254

Substitutedthiocarbamido) R.D. Isankar
Aminoindole by Microwave P.V. Raut
Technique
52 Understanding the Role of S. R. Deshmukh 255-258
Lipophilicity and Pharmacophore P.P. Nalawade
Elements in Pyrimidinedione based V. H. Masand
BCAT1 Inhibitory Activity S. D. Thakur
P. S. Navale
53 Use of Formic Acid as Co Shoeb R. Khan 259-264
Surrogates in Palladium Catalyzed Pratik E. P. Michael
Carbonylation Reaction Mayur V. Khedkar
54 In-Silico Prediction of Shweta Dhole 265-275
Phytoconstituents From Parimal Katolkar
Solanum Indicum for Antianxiety Pradeep Raghatate
Activity Jagdish Baheti
55 Computational Insight on the Sonali Ramgopal Mahule 276-279
Argentophilic Interactions of
Axially Chiral Ag-NHC Complex of
R-BINOL Scaffold via TD-DFT
56 Agricultural Influence and S. R. Kelode 280-283
application of Mn(III), Fe(III) and
VO(IV) Schiff base metal
Complexes
57 Recent Applications of Deep Suchita B. Wankhede 284-290
Eutectic Solvent (DES), a Greener
Media in Organic Synthesis
58 Synthesis, Characterization and Sushil K. Pagariya 291-297
Pharmacological Evaluation of Pravin S. Bodkhe
Some New Functionalized Flavone
Derivatives From Β-Diketones
59 Design, Synthesis, Spectral S. B. Sarkate 298-302
Characterisation and Antibacterial S. A. Waghmare
Screening of Some Novel 4- K. U. Dongare
R. N. Ingole
Substitutedimino-1,3,5-Dithiazine
Along With Pyrimidine Nucleus
60 Colorimetric Phytochemical Dr Swaroopa Rani N. Gupta 303-317
Analysis of Tinospora Cordifolia
and FTIR Study of Its Medicinal
Sample Giloy Ghanvati
61 Determination of Hydochloric Acid Dr Swaroopa Rani N. Gupta 318-328
Neutralizing Capacity of Different
Antacid Tablet by Back Titration
Method and Its FTIR Analysis
62 Investigation of Physico-chemical Dr. S. S Deshmukh 329-332
parameters of groundwater from Dr. Tejas R. Patil
Barshitakli tehsil region, Akola
district, Maharashtra, India
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

63 Identification of confluence of Vijay H. Masand 333-335

multiple scaffolds required for S.D. Thakur
Aurora Kinase A inhibition M.M. Rathore
64 Synthesis, Characterization and Dr. V. M. Sherekar 336-339
Biological Evaluation of 4-(4- Mr. Nilesh S. Padole
Bromo-1-Hydroxy Naphthalen-2-
Yl)-6-(4-Hydroxy Phenyl)-5,6-
Dihydropyrimidine-2(1h)-One
65 Efficient Synthesis of Vivek Ramkrushna Mate 340-345
Salicylaldehyde-Based Schiff Bases Datta Anandrao Patil
Using Hydrothermal Sonication: A Kalpana B. Gawande
Green Approach Sanjay R. Thakare
Rajkumar Uddhavrao
Pokalwal
66 Viscometric Technique for S.S. Padhen 346-348
Evaluation A.B. Wadekar
of 1-Phenyl-3-[4-(2-Ethylimino-4-
T-Butylimino-1,3,5-
Dithiazino)Amino- Phenyl]Prop-2-
Ene-1-One in Ethanol-Water
Mixture
67 An Overview on Green Chemistry Dr. Rupali.S. Talegaonkar 349-353
68 3D Printing Technology for Dr. Amar Deshpande 354-358
development of Transdermal Drug Dr. Jagdish Baheti
Delivery Systems
69 In-silico Prediction of Parimal Katolkar 359-372
Phytoconstituents from Jagdish Baheti
Hymenodictyon excelsum for
Antimalarial Activity Targeting
Hypoxanthine-guanine
phosphoribosyltransferase (HPRT1)
70 Spectrophotometric Complexation Santosh M. Chavan 373-377
Study of Cu(II) with 8- Nilesh V. Rathod
Hydroxyquinoline based Azo dye Manoj S. More
Chandrakant D. Ghugare
Ravi E. Khadse
Jayshri S. Jadhao
Akash V. Kubade
Parikshit S. Thakare
71 A Comprehensive Review on Rahul P. Rahate 378-383
Various Method of Synthesis and
Biological Activities of Schiff Bases
with Heterocyclic Moiety
72 Revolutionizing Applications: A S. R. Khandekar 384-389
Comprehensive Review of Schiff
Base Nanoparticles at the
Nanoscopic Scale
73 Screening Of Antioxidant Property V D Mane 390-394
Present In Psidium Guajava Linn P P Mhasal
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

74 In-Silico Prediction of Pooja P. Patle 395-405

Phytoconstituents From Solanum Parimal Katolkar
Indicum for Antiepileptic Activity Pradeep Raghatate
Jagdish Baheti
75 Development and Characterization Pragati Hasbe 406-410
of Piroxicam Matrix Based Zohra Firdous M.S.A
Transdermal Patch Pankaj Dhapke
Jagdish Baheti
76 Analytical Method Development Prajakta Sontakke 411-414
and Validation of Antidiabetic Disha Dhabarde
Drugs Jagdish Baheti
77 Synthesis of colloidal metal oxide Ramesh N. Zade 415-418
using reverse micelle technique and Pravin S. Bodakhe
its application in coupling reactions Kishor B. Raulkar
for Benzoxazole formation Bhupesh M. Mude
78 Synthesis, Charecterization and Disha M. Dhabarde 419-420
Biological Evaluation of Coumarin – Punam B. Rathi
Chalcone Derivatives Ashish Telrandhe

79 Synthesis and Characterization of P. S. Nandurkar 451-424

(3-(3,5-dichloro-2-hydroxyphenyl)- M. M Rathore
5-(pyridin-2-yl) isoxazol -4-
yl)(phenyl) methanone with
microwave irradiation
80 Microbial Examination of Ashish G. Sarap 425-428
Nanoparticle of Tetra-O-acetyl-B-D- P.T. Agrawal
Glucosyl-5aryl-4-Dithiobiurets
81 Synthesized PANI/Cu-NPs / Aloe- Dr. D. B. Dupare 429-435
Vera thin films Biocomposites for
ammine gas sensor stimulator
82 Structural and Ion-Exchange P. P. Kalbende 436-445
Properties of Copolymer Derived M. S. Dhore
from 4, 4’-Biphenol - Benzidine – S. S. Butoliya
Formaldehyde
83 Navigating the Therapeutic Mr. P S. Nawale 446-451
Landscape: Discovery and Ms. A D. Mehakare
Pharmacophore Modeling of BRD4 Dr. J R. Bansod
Inhibitors for Prostate Cancer Prof. Capt. M M. Rathore
Precision Treatment

*****
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

.
Chemistry
.
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

1
QSAR evaluation of Glucagon Receptor (GCGR) antagonists for the anti-
hyperglycaemic lead development

Ajaykumar Gandhi1, Pooja Gaikwad1, Jyoti Dahatonde1, Archana Chapolikar1*.

1
Department of Chemistry, Government College of Arts and Science, Aurangabad, Maharashtra, India, 431004
([email protected] , [email protected], [email protected], [email protected] )
*
Correspondence: [email protected], Mobile Number- (+91)9657538276.
ABSTRACT: GA-MLR based QSAR evaluation has been effectuated on a small dataset of GCGR
antagonists abide by OECD principles. Present study t revealed some of the structural features such as
minimum possible occurrence of -H2CCH-, PhCH2- moieties and more number of carbon atoms at
topological distance of 10 from halogen(s) as crucial for better GCGR antagonistic activity. Developed
QSAR model performed very well for all the approved statistical parameters such as, R2 = 0.90, Q2LOO
= 0.87, Q2LMO = 0.82, Q2Fn = 0.86–0.92, CCCext = 0.93. Applicability domain analysis and Y-
randomization test also enhanced the credibility of this study.
KEYWORDS: QSAR, GCGR, hyperglycaemia
ABBREVIATIONS:
CADD- Computer Aided Drug Designing, GCGR- Glucagon Receptor, GCGRA- Glucagon Receptor
Antagonist, SMILES- Simplified Molecular-Input Line-Entry System, GA- Genetic Algorithm,
MLR- Multiple Linear Regression, QSAR- Quantitative Structure-Activity Relationship, QSARINS-
QSAR Insubria, OECD- Organization for Economic Co-operation and Development, OFS- Objective
Feature Selection, SFS- Subjective Feature Selection
1. INTRODUCTION
A lifestyle induced metabolic disorder, Type 2 Diabetes Mellitus (T2DM) is the prevalent and ever
escalating type of the diabetes, worldwide. To date, 463 million people have diabetes and an estimated
global projection for diabetes is 578 million adults by 2030[1]. Cardiovascular discomfort, retinopathy,
nephropathy are some of the health complications that are consequences of hyperglycaemia. Glucagon
– a peptide hormone, known to elevate hepatic glucose production by stimulating gluconeogenesis and
glycogenolysis and regulates blood glucose homeostasis collaboratively with insulin. In both, type 1
and type 2 diabetes, increase in glucagon level and/or insufficient glucagon suppression contribute to
the hyperglycaemia. Glucagon Receptor (GCGR) antagonism emerged as a promising therapy to
control hyperglycaemia. BAY 27-9955 was the first clinically approved GCGR antagonist. MK-
357732, MK-0893, PF-06291874, LY- 2409021, and LGD-6972 under trial GCGR antagonist[2]–[5].
GCGR is a well-established target for glucagon suppression and hence further optimization of reported
compounds may possibly suffice a better lead of desired anti-hyperglycemic potency with minimum
and less serious side effects
Computer Aided Drug Designing (CADD)[6]–[8] being result oriented, time- and cost-
efficient, and eco-friendly approach, is now an established drug discovery approach. QSAR–is a
multidisciplinary approach wherein experimentally determined bio-activity of molecule is statistico-
mathematically correlated to its molecular features (expository QSAR) which further assist in an in
silico bio-activity prediction (statistical QSAR)[9]–[11]. Expository QSAR intelligibly aligned with
statistical QSAR provides better insights for the pharmacokinetics[12]–[19].
In the recent work, a QSAR model using dataset of 44 GCGRAs with experimentally
determined GCGR binding activity is developed. Decidedly this model will assist synthetic organic
chemists in the development of better GCGRA as anti-hyperglycemic lead.

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

2. MATERIALS AND METHODS

A. Selection of data-set:
A dataset of 44 Glucagon Receptor Antagonists (GCGRAs) with in vitro GCGR binding
activity in terms of half-maximal inhibitory concentration (IC50) ranging from 0.02 to 131.1 μM
(converted to pIC50) is used[20], [21]. The variation in GCGR binding activity with chemical features,
illustrated using five least and five most active compouds (Figure. 1). The SMILES strings with
reported IC50 and pIC50 values for all the GCGRAs are present in the Table S1 in the supplementary
material. (A google drive for supplimentary information provided at the end)
B. Molecular Structure Drawing and Optimization:
Free and open source software, ChemSketch 12 Freeware (www.acdlabs.com) is used to draw
2D structures and further conversion to 3D structures is achieved using OpenBabel 2.4.0. A geometry
optimized molecule is characterized by its lowest energy i.e. most stable conformation. Geometry
optimization ensures the normalization of all such a physico-chemical properties for all the molecules
from dataset and makes it a necessarly important step prior to molecular descriptor calculation. As a
step forward, force field MMFF94 available in TINKER is used for optimization of the molecules.

Figure 1: Variations in activity and chemical structure in the present dataset of GCGRAs (a) five least active
compounds (b) five most active compounds.

C. Molecular Descriptor Calculation and Objective Feature Selection (OFS):

A molecular descriptor is a structural and physico-chemical property of a molecule or specific
part of the molecule. More than 18,000 molecular descriptors were calculated for each molecule using
PyDescriptor[22] and PaDEL[23]. In the data pruining step, Objective Feature Selection (OFS) in
QSARINS v2.2.4[24], [25] screened out multi-collinear and spurious molecular descriptors (i.e. with
nearly constant values >95%, co-linearity |R| >0.95) and contracted molecular descriptor pool with 1145
variables is generated. Contracted though, molecular descriptor pool has covered adequately
comprehensive chemical space being comprised of 0D– to 3D– descriptors, structural, constutional
properties and charge descriptors etc.
D. Subjective Feature Selection, QSAR Model – Development and Validation:
In accordance with the OECD principles, firstly, apt variable selection method GA– MLR[26]
in Subjective Feature Selection (SFS) operation in QSARINS v2.2.4 is used to perform simple and
easy to interpret QSAR models. Then, all the derived models were subjected to the thorough

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

statistical validation, Y-scrambling and Applicability Domain analysis. The steps in QSAR building
process:
I. Random split operation in QSARINS v2.2.4 split given dataset into a training set with 35
molecules (80%) and a prediction set with 9 molecules (20%) prediction. Molecules in training
set were utilized for QSAR model development and external validation was performed on 9
molecules in prediction set.
II. QSAR models were built using Subjective Feature Selection (SFS) operation in QSARINS
v2.2.4 (at default settings) by setting Q2LOO as fitness function. Insignificant increase in Q2LOO
value was observed after 5 variables and hence to avoid overfitting SFS opertion is confined
to 5 variables which additionally helped in deriving easy and informative QSAR models. (See
supplementary information Table S2)
III. (a) Leave-One-Out (LOO) and Leave-Many-Out (LMO) parameter based internal validation;
(b) External validation; (c) Y-scrambling and model Applicability Domain (AD) analysis,
performed for legitimate validation.
IV. Performance of each model, measured by close inspection of the various statistical parameters
meter the robustness of the GA-MLR based QSAR model. The QSAR model with best values
of these parameters and with best predicative ability is selected.
3. RESULTS
 Values of Fitting, Internal Validation, and External Validation are very well above the
approved threshold values.
 The predictiveness of the generated QSAR model can be corroborated from Figure 2a.
 Moreover, Model applicability domain (AD) of the model is corroborated from Williams
plot (Figure.2b).
 QSAR Model (Divided Set: Training Set-80% and Prediction Set-20%):
pIC50 = 7.941(±1.078) - 0.148(±0.068)H_don_8B - 1.594(±0.204)KRFPC298 -
1.700(±0.490) KRFPC582 + 0.106(±0.047) APC2D10_C_X
[R2 = 0.90, R2adj = 0.88, Q2LOO = 0.87, Q2LMO = 0.82, RMSEtr = 0.25, MAEtr = 0.25, RSStr
= 3.86, CCCtr = 0.95, s = 0.35, F = 70.22, RMSEcv = 0.36 MAEcv = 0.29, PRESScv = 4.80,
CCCcv = 0.93, R2ext = 0.90, Q2-F1 = 0.88, Q2-F2 = 0.86, Q2-F3 = 0.92, CCCext = 0.93]
 Compounds 13, 40, 22, 17 and 16 with Predicted Models Equation Residual values in the
range –0.012 to +0.032 are best predicted for their GCGR activity (pIC50) in the present
QSAR evaluation (Figure. 3).
 All these five compounds fall within Applicability Domain, as evident from William Plots
(Figure. 2b). On the other hand, GCGR binding activity of Compound 31 is worst predicted
for by the QSAR models and turned out as an outlier.

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Figure 2: For QSAR Model (a) Graph of Experimental vs. Predicted pIC50 values (b) Williams plot.

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Figure 3: Depiction of first 5 Best Predicted Molecules

4. DISCUSSION
 H_don_8B (-VE correlation with GCGR binding potency) small possible number of Hydrogen
atoms within 8 bonds from HBD atom is advisable for more potent GCGR antagonist
 KRFPC298 and of KRFPC582 8B (-VE correlation with GCGR binding potency) less frequent
occurrence of H2C-CH and of PhCH2- structural fragments, respectively are recommended for
better GCGR binding potency.
 APC2D10_C_X (+VE correlation with GCGR binding potency) more frequent occurrence of
Carbon at topological distance of 10 from any halogen is advocated by QSAR evaluation

5. CONCLUSIONS
Decidedly, a QSAR evaluation of a given series of compounds revealed some prominent
structural/chemical traits that are responsible for the enhanced activity of the compound, such as
minimum occurrence of -H2CCH- and PhCH2- structural fragments, and less possible number of
hydrogen atoms within eight bonds from hydrogen bond donor atoms, and more number of Carbons at
topological distance of ten from halogen(s) which revealed the scope for optimization of the lead for
better GCGR binding potency. The QSAR model will assist in optimizing leads to better GCGR binding
potency to curb hyperglycaemia and diabetes.

ACKNOWLEDGMENTS: The authors are thankful to Dr. Paola Gramatica and her team for
providing QSARINS-v2.2.4 and developers of TINKER, ChemSketch 12 Freeware (ACD labs), and
PyDescriptor for providing the free versions of their software.

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REFERENCES:
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[Online]. Available: https://diabetesatlas.org/en/.
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glucagon receptor antagonist LGD-6972 in healthy subjects and subjects with type 2 diabetes mellitus,”
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dichlorophenyl)-5-(6-methoxynaphthalen-2-yl)-1 H -pyrazol-1-yl]ethyl}phenyl) carbonyl]-β-alanine
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‘Push a button and find a correlation’: A case study of toxicity of (Benzo-)triazoles on Algae,” Mol.
Inform., vol. 31, no. 11–12, pp. 817–835, 2012, doi: 10.1002/minf.201200075.
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[18] V. H. Masand, D. T. Mahajan, G. M. Nazeruddin, T. Ben Hadda, V. Rastija, and A. M. Alfeefy, “Effect
of information leakage and method of splitting (rational and random) on external predictive ability and
behavior of different statistical parameters of QSAR model,” Med. Chem. Res., vol. 24, no. 3, pp. 1241–
1264, Aug. 2015, doi: 10.1007/s00044-014-1193-8.

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[19] P. Gramatica, “On the development and validation of QSAR models,” Methods Mol. Biol., vol. 930, pp.
499–526, 2013, doi: 10.1007/978-1-62703-059-5_21.
[20] S. Shu et al., “A novel series of 4-methyl substituted pyrazole derivatives as potent glucagon receptor
antagonists: Design, synthesis and evaluation of biological activities,” Bioorganic Med. Chem., vol. 26,
no. 8, pp. 1896–1908, 2018, doi: 10.1016/j.bmc.2018.02.036.
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containing derivatives as a novel series of potent glucagon receptor antagonists,” Bioorganic Med.
Chem., vol. 24, no. 12, pp. 2852–2863, 2016, doi: 10.1016/j.bmc.2016.04.053.
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models for environmental pollutants in QSARINS,” J. Comput. Chem., vol. 35, no. 13, pp. 1036–1044,
May 2014, doi: 10.1002/jcc.23576.
[25] P. Gramatica, N. Chirico, E. Papa, S. Cassani, and S. Kovarich, “QSARINS: A new software for the
development, analysis, and validation of QSAR MLR models,” J. Comput. Chem., vol. 34, no. 24, pp.
2121–2132, Sep. 2013, doi: 10.1002/jcc.23361.
[26] A. K. Saxena and P. Prathipati, “Comparison of MLR, PLS and GA-MLR in QSAR analysis,” SAR
QSAR Environ. Res., vol. 14, no. 5–6, pp. 433–445, 2003, doi: 10.1080/10629360310001624015.
[27] K. Roy, S. Kar, and P. Ambure, “On a simple approach for determining applicability domain of QSAR
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[28] T. M. De Assis et al., “QSAR Models Guided by Molecular Dynamics Applied to Human Glucokinase
Activators,” Chem. Biol. Drug Des., vol. 87, no. 3, pp. 455–466, 2016, doi: 10.1111/cbdd.12683.
[29] P. Gramatica, “Principles of QSAR models validation: Internal and external,” QSAR Comb. Sci., vol. 26,
no. 5, pp. 694–701, 2007, doi: 10.1002/qsar.200610151.
[30] A. Arwansyah, A. R. Arif, G. Syahputra, S. Sukarti, and I. Kurniawan, “Theoretical studies of
Thiazolyl-Pyrazoline derivatives as promising drugs against malaria by QSAR modelling combined
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https://doi.org/10.1080/08927022.2021.1935926, 2021, doi: 10.1080/08927022.2021.1935926.

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2
Synthesis and Bilogical Evaluation of Pyrazole Derivative
1A P Thakare & 2S R Kolteke
1
Department of Chemistry, Rajarshee Shahu Science College, Chandur Rly
2
Mahatma Fule Arts, Commerce & Sitaramji Chaudhari Science College, Warud
Email- [email protected]
Abstract:
Pyrazole, a five-membered heterocycle containing two nitrogen atoms, is extensively found as
a core framework in a huge library of heterocyclic compounds that envelops promising agro-
chemical, fluorescent and biological potencies. Attributed to its several potential applications,
there is a rise in the significance of designing new pyrazoles. The current study presents the
synthesis of pyrazole derivatives by the reaction of chalcones with aryl hydrazine
hydrochlorides in acetic acid (30%) under reflux conditions in good yields. Structures of
synthesized new pyrazoles were confirmed by spectral studies.
Biological activity of all the synthesized compounds was checked against gram positive and
gram negative bacteria. It has been found that all compounds shows good antimicrobial
activity.
Introduction:
Heterocyclic compounds are a highly valuable and unique class of compounds. These
compounds demonstrate a broad spectrum of physical, chemical and biological characteristics.
Amongst heterocyclic compounds, nitrogen-containing heterocycles are extensively found as
a core framework in a huge library of heterocycles and show several employments in natural
science and other areas of science. Additionally, nitrogen-containing heterocycles have
striking structural features and they are widely observed in natural products, for instance,
vitamins, hormones and alkaloids.
Chalcones are the principal precursors for the synthesis of bioactive small molecules such as
benzothiazepines, pyrazolines, isoxazolines, cylopropanes, oxadiazoles, etc., The chalcones
are most commonly synthesized via Claisen-Schmidt reaction of an aromatic aldehyde with
acetophenones. Chalcones has gained importance due to their simple structures and diverse
pharmacological applications. Design and synthesis of simple heterocycles with various
bioactivities is a worthwhile contribution in organic synthesis. The compounds with thiophene
are the most important class in active pharmaceutical drugs and remain the choice for anti-
inflammatory agents in spite of multiple attempts at exploring alternative scaffolds. [1-5]
II. EXPERIMENTAL
All melting points were determined in open glass capillaries and are uncorrected. The IR
spectra were recorded on KBr disc using Perkin Elmer-1800 intrachord. 1HNMR and 13C NMR
spectra were recorded in CDCl3 on Brucker Avance 400MHz spectrophotometer with TMS as
internal standard (chemical shifts are expressed in δ ppm). The mass spectra were recorded on
a Joel SX-102 (EI/CI/FAB) mass spectrometer at 70 eV. The reactions were monitored by the
TLC on silica gel G plates in the solvent system benzene–methanol mixture (9:1). All reagents
were purchased from commercial suppliers and used without further purification. The
compound includes 2-hydroxy-4-methylacetophenone, benzaldehyde, p-chloro benzaldehyde,
anisaldehyde, 4-methyl benzaldehyde.
Synthesis of chalcone:
A mixture of 0.01 mol 2-hydroxy-4- methylacetophenone and 0.01 mol various aldehyde added
into ethanol solvent. To this reaction mixture 20 % NaOH added and heated for several minutes

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upto formation of solid residue. By keeping overnight residue nuetralized by ice cold HCl
solution, filtered and dried in oven.

Synthesis of Pyrazole:
A mixture of synthesized chalcone and aryl hydrazine hydrochloride in aqueous acetic acid
under reflux condition produced pyrazole derivative.

Result & Discussion:

Sr No Compounds R1 R2 R3
1 2-(1,5-diphenyl-1H-pyrazol-3-yl)phenol H H H
2 2-(1-phenyl-5-(p-tolyl)-1H-pyrazol-3-yl)phenol CH3 H H
3 2-(5-(4-chlorophenyl)-1-phenyl-1H-pyrazol-3- Cl H H
yl)phenol
4 2-(5-(4-methoxyphenyl)-1-phenyl-1H-pyrazol-3- OMe H H
yl)phenol
5 2-(5-(4-methoxyphenyl)-1-phenyl-1H-pyrazol-3- OMe CH3 H
yl)-5-methylphenol
6 2-(5-(4-chlorophenyl)-1-phenyl-1H-pyrazol-3-yl)- Cl CH3 H
5-methylphenol
7 5-methyl-2-(1-phenyl-5-(p-tolyl)-1H-pyrazol-3- CH3 CH3 H
yl)phenol

Table: Analytical Data of Synthesized Compounds

Sr No Molecular M.P. (oC) Yield (%) C% H% N%
formula
1 C21H16N2O 112 84 80.75 5.16 8.97

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2 C22H18N2O 119 78 80.96 5.56 8.58

3 C21H15ClN2O 110 76 72.73 4.36 8.08
4 C22H18N2O2 123 81 77.17 5.30 8.18
5 C23H20N2O2 121 84 77.51 5.66 7.86
6 C22H17ClN2O 120 80 73.23 4.75 7.76
7 C23H20N2O 105 88 81.15 5.92 8.23

1) 2-(1,5-diphenyl-1H-pyrazol-3-yl)phenol

IR (KBr) ν: 3059(Ar C-H), 3345 (O-H), 1330 (C-N), 1226 (C-O).

1
H-NMR (400 MHz, CDCl3) δ (ppm) 7.05 (s, 1H, OH), 7- 8.26 (m, 14H, Ar)
2) 2-(1-phenyl-5-(p-tolyl)-1H-pyrazol-3-yl)phenol

IR (KBr) ν: 3059(Ar C-H), 3345 (O-H), 2990 (aliphatic C-H), 1328 (C-N), 1226 (C-O).
1
H-NMR (400 MHz, CDCl3) δ (ppm) 7.05 (s, 1H, OH), 7- 8.26 (m, 13H, Ar), 2.34 (s, 3H,
CH3)
3) 2-(5-(4-chlorophenyl)-1-phenyl-1H-pyrazol-3-yl)phenol

IR (KBr) ν: 3059(Ar C-H), 3345 (O-H), 2990 (aliphatic C-H), 1328 (C-N), 1226 (C-O).
1
H-NMR (400 MHz, CDCl3) δ (ppm) 7.02 (s, 1H, OH), 7- 8.26 (m, 13H, Ar)
4) 2-(5-(4-methoxyphenyl)-1-phenyl-1H-pyrazol-3-yl)phenol

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IR (KBr) ν: 3059(Ar C-H), 3345 (O-H), 2990 (aliphatic C-H), 1328 (C-N), 1226 (C-O).
1
H-NMR (400 MHz, CDCl3) δ (ppm) 7.02 (s, 1H, OH), 7- 8.26 (m, 13H, Ar), 3.81 (s, 3H,
OCH3)
5) 2-(5-(4-methoxyphenyl)-1-phenyl-1H-pyrazol-3-yl)-5-methylphenol

IR (KBr) ν: 3059(Ar C-H), 3345 (O-H), 2990 (aliphatic C-H), 1328 (C-N), 1226 (C-O).
1
H-NMR (400 MHz, CDCl3) δ (ppm) 7.02 (s, 1H, OH), 7- 8.26 (m, 12H, Ar), 3.81 (s, 3H,
OCH3), 2.28 (s, 3H, CH3)
6) 2-(5-(4-chlorophenyl)-1-phenyl-1H-pyrazol-3-yl)-5-methylphenol

IR (KBr) ν: 3059(Ar C-H), 3345 (O-H), 2990 (aliphatic C-H), 1328 (C-N), 1226 (C-O).
1
H-NMR (400 MHz, CDCl3) δ (ppm) 7.02 (s, 1H, OH), 7- 8.26 (m, 12H, Ar), 2.28 (s, 3H,
CH3)
7) 5-methyl-2-(1-phenyl-5-(p-tolyl)-1H-pyrazol-3-yl)phenol

IR (KBr) ν: 3059(Ar C-H), 3345 (O-H), 2990 (aliphatic C-H), 1328 (C-N), 1226 (C-O).
1
H-NMR (400 MHz, CDCl3) δ (ppm) 7.02 (s, 1H, OH), 6.89- 7.55 (m, 12H, Ar), 2.28 (s, 3H,
CH3)
Biological Activity
Compounds were screened for their antibacterial and antifungal activity using cup-plate agar
diffusion method at a concentration of 40 mg, using Gram positive bacterial strains such as B.
cocous and B.subtillus and Gram negative bacterium strain such as Proteus vulgaris and
Escherichia coli. The antifungal testing was carried out against Aspergillus niger. Known
antibiotics like Amoxycillin, Benzoylpenicillin, Ciprofloxacin, Erythromycin, and antifungal
activity was compared with Greseofulvin. The zone of inhibition measured in mm. By

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visualizing the antimicrobial data, it could be observed that most of the compounds exhibited
significant activity.

References:
1. Manjunath B.C., Manjula M., Raghavendra K.R., Ajay Kumar K., Lokanath N.K. (2014) 4-(Thiophen2-yl)-
2-[4-(trifluoromethyl)-phenyl]-2,3-dihydro-1,5-benzothiazepine. Acta Cryst. Sect. E, 70 (Part 3).
2. Kumar G.V., Govindaraju M., Renuka N., Khatoon B.B.A., Mylarappa B.N., Kumar K.A. (2012) Synthesis
of 1,3,5-triaryl-4,6-dioxo-pyrrolo[3,4-d]-7,8-dihydropyrzoles and their antimicrobial and antioxidant
activity. Rasayan J. Chem. 5 (3) 338–342.
3. Ajay Kumar K., Govindaraju M., Vasantha Kumar G. (2010) Synthesis of isoxazoles via 1,3-dipolar
cycloaddition reactions and their antimicrobial activity. Ind. J. Heterocycl. Chem. 20 (4) 183-184.
4. Ajay Kumar K., Lokanatha Rai K.M., Vasanth Kumar G., Mylarappa B.N. (2012) A facile route for the
synthesis of ethyl N-aryl-2,6-dioxo-piperid-3-ene-4-carboxylates and their biological activity. Int. J. Pharm.
Pharm. Sci. 4 (Suppl 4) 564-568.
5. Ajay Kumar K., Lokanatha Rai K.M. (2004) Synthesis and evaluation of antimicrobial activity of 4,5-
dihydro-12,4-oxadiazoles, Bulg. Chem. Commun. 36 (4) 249-252.
6. Chikulla, K V. Raja, S. Isoxazole-A Potent Pharmacophore, I. J. of Pharmacy and Pharmaceutical Sci., 9(7):
13-24.
7. Jyothi, A N. Poojitha, J. Raju, G N. Nadendla, R R. Potential activities of isoxazole derivatives, World J. of
Pharmaceutical Research. 4(12): 667-679.
8. Dou, G. Xu, P. Li, Q. Xi, Y. Huang, Z. Shi,D. 2013. Clean and Efficient Synthesis of Isoxazole Derivatives
in Aqueous Media, Molecules, 18: 13645–13653.

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3
Healthy hydroponic cattle feed (grass) development eco-friendly &
economically

Dr. Ramesh Tukaram Parihar, Dr A.G.Gaddamwar*

Department of chemistry, Vidnyan Mahavidyalaya Malkapur, Dist-Buldhana, Affiliated to Sant Gadge Baba
Amravati University Amravati, Maharashtra, India
Department of chemistry, Amolakchand Mahavidyalaya Yavatmal, Dist-Yavatmal, Affiliated to Sant Gadge
Baba Amravati University Amravati, Maharashtra, India

Abstract
Dairy cattle require green fodder for high milk yield. However, it cannot available
throughout the year and in some area, it is difficult to have access for green fodder. Thus,
hydroponic fodder production has become an alternative way to fulfil this green fodder
requirement of the dairy cow in low cost. According to WHO the production of milk in the
India is 14 crore litres, but the consumption is 64 crore litres. It is being claimed that 68.7
percept of the milk and its products sold in the country are adulterated. In the US, the average
dairy cow produces more than 7.5 gallons of milk per day and India’s milk processing capacity
is 126 million litres per day, the highest in the world, and lauded the dairy sector for increasing
production from 22 MMT, or around 6 crore litres per day in 1977, to 58 crore litres per day in
2022. The adoption of this technique has enabled the production of fresh forage from
grains without soil. Hydroponic fodder has high nutritive value due to the conversion of
complex compounds into simpler and essential form, and activation of enzymes during
germination. Thus, it contains high protein, vitamins and minerals which are essential for dairy
cows. There were improvements in digestibility and intake of nutrients results in increased milk
yields and quality like milk fat of dairy cow on the feeding of hydroponic fodder. Traditional
fodder production has a number of limitations regarding soil & climatic conditions. In
successful mulch animal rearing green fodder has its special significance.
Keywords: Hydroponics, Eco-friendly, Economics, nutritional value, Amino Acid.
Introduction
The word hydroponics has been derived from the Greek word “water working”. Hydro means
“water” and phonic means “working” and it is a technology of sprouting grains or growing
plants without soil, but only with water or nutrient rich solution. However, hydroponics fodder
can be well produced with the use of fresh water only It is one of the most important agricultural
techniques currently in use for green forage production in many countries especially in arid
and semi-arid regions environmentally control houses. The population of Dairy livestock has
increased immensely in our country, but still it is unable to fulfil the required milk Demand
that we need to import it. Dairy cattle require green fodder for being healthy and higher Milk
yield. The facts that deficiency of feed and fodder alone Account nearly 50 percept losses in
livestock. To overcome these hydroponic cattle feed cultivation is introduced. Hydroponics
fodder production has two aspects i.e. Physiology and nutrition of plant-animal system and
Engineering of hydroponic technology. Hydrophophic is the science of soilless growing of
plants in Nutrient rich solutions at regulated temperature and humidity. The main problems of
feed scarcity emanate from land scarcity; actually, rapid urbanization is the major cause behind
the decrease in land meant for grazing and fodder cultivation. With Water, labour shortage and
elevated cost of fertilizers the farmer leans to cultivate commercial food crops over green
fodder. Nevertheless, producing green fodder to meet the current demand has become a greatest
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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

challenge among livestock farmers. In fact, green fodder is very important for productive and
reproductive performance of animals.
Feeding green fodder can improve livestock products. Livestock production in most regions is
limited due to poor production and pricy imported green fodder. Today, land scarcity presents
an important limit towards forage production for animal especially sheep, goats and cattle.
Unlike monogastric mammals, ruminant cannot solely dependent on cereal grains. That’s why,
alternative technologies, such hydroponics, are regarded as vital to face these issues. The use
of this technology can help improve the long-term economic development of the livestock
industry. Hydroponic fodder can also improve the performance of the animals by providing
their nutrient needs. Hydroponic growing green fodder has high feed quality, rich with proteins,
fibres, vitamins, and minerals effects on animals. The growth of the fodder crop mainly
depends on moisture, temperature, RH and irrigation. Hydroponic Green fodder is the natural
diet for livestock. Hydroponic green fodder consists of grass with grains, roots, stems and
leaves which are highly nutritious and provide sustainable fodder production and conserve
water take 8 days to develop seed to green fodder. It’s production to meet current demand has
become a greatest challenge among livestock farmers, due to temperature, humidity and
contamination on the seed and fodder are restrict the optimum growth of fodder. Amount of
yield and quality of fodder is influenced by grain quality, grain variety and treatments and
growing environmental. Green fodder production crisis serious problems which are
contamination of seeds that effect germination, and plant growth. This contamination further
takes to fungus and mould infection which are not healthy for livestock animals. Temperature,
relative humidity is also parameter to increase and decrease the fungus and mould infection.
Hydroponic farming means growing of plants without soil by using nutrients water at desired
temperature and humidity. Through hydroponics it is easy and quick to produce nutritive green
fodder. Green fodder is the natural diet for livestock. Its production to meet the current demand
has become a greatest challenge among livestock farmers. Due to many reasons, green fodder
production has been facing a serious crisis and so the livestock productivity. Due to increasing
intensive system of rearing livestock, the need for green fodder is enormous .As the gap
between the demand and supply of the green fodder for livestock becoming unconquerable,
researchers and farmers are in search for an alternative fodder or fodder production method,
that would restore fodder and livestock production.
Hydroponics is the state of the art technology that has revolutionised the green fodder
production in the 21st century. Hydroponics is a method of growing green fodder without soil
in an environmentally controlled houses or machines. Many of the livestock farmers are
switching to hydroponic fodder production from conventional production methods, as the
fodder produced by this method are highly nutritious, provide sustainable fodder production
round the year and conserve water. Though this method has made a greatest impact in the
fodder production system, most of the farmers are facing some practical difficulties in
profitably running the hydroponic machine for sustainable fodder production. This manual has
been compiled with the essential manage mental practices that have to be carried out for an
economically sustainable fodder production. Green fodder plays major role in feed of all
livestock. Also, into the milch animals, Green fodder providing required nutrients for milk
production and health of the dairy animals. Green fodder feeding to livestock is important for
optimization of productivity. Animals Feeding and fodder production are the two important
aspects for the sustainability of products and productivity in animal rearing. Although, India is
the top producer of milk in the world But there are many challenges insufficient livestock feed,
fodder is one of the constraints Affecting growth, health, production and reproduction potential
of livestock. Green fodder is the natural diet for livestock. Its production to meet the current
demand has become a greatest Challenge among livestock farmers. Due to many reasons, green

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fodder production has been Facing a serious crisis and so the livestock productivity. Due to
increased population day by Day hence agriculture land is converted into urbanization. Now
days small land holdings Amongst farmers, non-availability of irrigated lands for fodder
production is reduced, Unavailability of fertile land for fodder production, increasing mining
and coastal line has Limited area for fodder production, deforestation, lack of scientific
knowledge of feed and Fodder production among unemployed youths for fodder farming,
higher labour cost and Small land holdings has left livestock as well as dairy farmer with many
challenges for Animal rearing and milk production in all over the world. Also, due to increasing
intensive System of rearing livestock, the need for green fodder is huge. To overcome all these
shortage Issue of green fodder the new hydroponics technology came into exist. Hydroponics
is the state-of-the-art technology that has revolutionised the green fodder production in the 21st
century. In India only 4.9% of cropped land area is utilized for cultivating fodder. Indian
livestock industry faces a deficit of 35.6% green fodder, 26% of dry fodder and 41% of
concentrate feed ingredients.

Problem statement
A suitable combination of green & dry fodder is Very important for maintaining animal
health& milk production. But in Scarcity condition traditional green fodder production become
impossible Because of lack of irrigation water. In such condition the technique of Green fodder
production by Hydroponic method is very useful tool. This technique does not require soil.
Hence limitations like Saline soil, inferior soil, water logged soil etc can be easily overcome.
This Technique requires very less quantity of water. Hence it can be easily undertaken in
scarcity affected areas. Green fodder is the natural diet of cattle. Green fodder is the most
Viable method to not only enhance milk production, but to also bring about A qualitative
change in the milk produced by enhancing the content of Unsaturated fat,, Omega 3 fatty acids
, vitamins, minerals and carotenoids. Hydroponics fodder growing is the state-of-the-art
technological Intervention to supplement the available normal green fodder resources required
by the dairy cattle. The contamination was starts from the seeds. When seeds were soaking for
24 hours in water there was problem of fermentation and because of that the fermented odour
take place to the seeds which are resulted the fungus infection. After soaking the seeds, the
seeds were placed into gunny bags at dark rooms for sprouting this resulted increase in
fermentation odour. Sprouted seeds when placed into the hydroponic or office tray water
sprayed to the seeds were for 2 minutes which are 1liter/min and because of that the maize
seeds had higher amount of water which are not suitable for fodder and the water are not well
drained from the trays this causes increase in fungus infection. Uncontrolled environment was
also helping to increasing the fungus infection if there is temperature is greater than 35 °c and
humidity higher than 70 to 80 % the fungus and mould infection were increases.
Materials and Method/procedures
Hydroponics fodder unit is nothing but a chamber with arrangement of temperature, humidity
and light intensity for maximum sprouting & growth of fodder crop seeds (mainly maize, oat,
barley, wheat etc). With 30 to 35 ˚C temperature, 60 to 75 % R humidity and 50 % shed 1 Kg
of maize should yield 6 to 8 Kg green fodder in 7 to 9 days. It found that for 6-8 times mass
increase around 2 litre water is sufficient per kg of seeds during summer season. So by this one
can grow very good, healthy & economical viable fodder for dairy / goat farming. (We suggest
to calculate economic output of system at your end based on input seed cost) There are plenty
of manufacturer suppliers of hydroponics systems in market but farmers can build their own
system as it’s very easy to fabricate & all components available in market.
Following are required components with their specifications -

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System components – Growing cabinet, Racks for trays, trays, seeds, watering system (fogger
/ mister / drip pipes) , timer, motor. (Photo look like are given below)
Growing cabinet – Square shape cabinet is most suitable & easy to fabricate , cabinet size can
vary based on number of trays , cabinet can be fabricated with mild steel pipes (MS) , UPVC
or bamboo. Just make square box and trays arrangement. Slotted angle racks can also be serve
purpose. Cabinet need to be covered with 50 % green shed net and/or poly house covering film
for reducing light intensity & conserves humidity.
Racks for trays – Can be of MS, UPVC or bamboo. Height of the tray racks need to be arranged
as per day cycle of fodder i.e lower level 2 racks with 6 inch , above 2-3 racks on 8 to 12 inch
and upper level racks 12 to 15 inch height. A gentle slope is beneficial for avoiding water
lodging and fungal growth.
Trays – Various kind of trays are available in market based on quality of plastic & durability,
specialized hydroponics trays are costly ( 2 feet * 2 feet tray cost approx. Rs.350) . We can use
simple office tray (1.5 feet *1 feet) with perforations at bottom (simply drill holes on equal
distance). Make sure there are sufficient holes made to avoid any kind of water lodging and
avoid fungal infection. After every use, trayneed to be disinfected by diluted hydrogen peroxide
and sun drying.
Seeds – Maize, wheat, oat, barely suits best for hydroponics. You can choose based on
availability and rate per unit. Seed should be free from any fungal infection. To remove broken
seed, give brine water treatment and remove floating seeds. Soak seed further hours in
potassium permanent. Once seeds spouted 2 treatments of powder tricoderma Species will also
help in lowering fungal Infection. Seed rate of 0.5 Kg / Ft2 is sufficient for hydroponics fodder
cultivation.
Watering system – Motor pump of 0.25 or 0.5 Hp is sufficient for 100 tray system. For lesser
Capacity, minimum 0.25 Horse power motor will be required for operating fogger / misters.
We prefer Jain Irrigation misters than foggers as they required less pressure & easy for
maintains. Drip line – regular 16 mm HDEP drip line is best suited.
Timer – Various timers available in market. Timer best suited are generally 1 to 2 min operation
Every / hour. We prefer Frontier TM-619-H-2 with 17 time intervals. If motor pump is above
3 Amp Additional really is preferable.
Engineering Aspects of Hydroponic Technology – Research in the following areas of
engineering and machinery development will help in making the system more acceptable to
the farmers.
Reservoir Engineering and Nutrient Solutions- The reservoir is the part of the hydroponic
System that holds the nutrient solution. Depending on the type of hydroponic system, the
Nutrient solution can be pumped from the reservoir up to the growing chamber (root zone) in
cycles using a timer, as well as continually without a timer. Reservoir can be made out Of
materials including the plastic that can hold water. Nutrient solutions can be developed
separately for hydroponic fodder crop and sprouted grain crop.
Delivery System-The delivery of the hydroponic system’s nutrient solution/water can be
customized as per local needs.
Submersible Pump- Most hydroponic systems use a submersible pump to pump water/ Nutrient
solution from the reservoir up to the growing chamber/root zone of the plants.

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Air Pump- Other than in water culture systems, air pumps are optional in hydroponic Systems.
However, using them has benefits. It helps to increase dissolved oxygen levels in the water up
and keep the water oxygenated. This helps in keeping the nutrients evenly mixed all the time.
Grow Lights- Grow lights are optional part of hydroponic systems. One can choose to either
Use natural sunlight or artificial light to grow the plants depending on the place of operation
of the hydroponic system. These are generally used in commercial hydroponic systems for
Growing fruits, vegetables and flowers.

Conclusion:
High initial investment on fully automated commercial hydroponic systems and high labour
and energy costs in maintaining the desired environment in the system adds substantially to the
net cost of hydroponic fodder production. Such systems are not successful in developing
countries. Conversely, low cost hydroponic systems have been developed by utilizing locally
available infrastructure where there is an acute shortage of fodder and water; local irrigation
systems are not well established; transportation and fuel costs are high; and seasonal variations
of fodder prices are extreme. Typical lean periods of fodder production are the norm,
investment in controlling temperature and humidity are low, and so is the cost of labour. Under
such situations the cost structure is often shifted in favour of hydroponic fodder production,
and it may find a niche in increasing livestock production. Hydroponics fodder is nutritious,
palatable and digestible and can be grown in low cost techniques with locally home grown
grains. Against impeding climate change and less availability land hydroponics fodder
production is an effective alternative technology for sustainable livestock production in
different agro climatic regions of India. The use of hydroponic culture modified the chemical
composition of milk with respect to fat content, which is a desirable parameter. Moreover,
principal component analysis revealed that with respect to proximate analysis, the quality of
milk from cows from the GZ farm was superior to that of cows from the MT farm. The
difference between minerals in the two farms may not be easily interpreted as the environment
plays an important role, particularly with contaminants in the air, water, and soil. Further
studies are needed to establish the effects of long-term feeding with different types of
hydroponic fodder and to investigate the effects of these hydroponic fodders on the productive
and reproductive performance of dairy cows. Further studies may be directed towards the
development of feeding strategies with respect to the inclusion of hydroponic fodder under
different agro climatic conditions. The challenges of Hydroponic fodder-cum-sprouted grain
technology are enormous in producing quality green feed and fodder, and using it for value
addition of products, such as milk rich in CLA, Omega-3, vitamins and micro minerals; cage-
free poultry products, grass-fed meat and meat products. Better outcome will be possible with
the strong policy support and technology back up. When used as supplement in the feeding of
dairy animals, it will improve health and reproduction; boost productivity as well as nutritional
quality of milk and other foods of animal origin, thus improving the income and profits of the
farmers. However, before adoption of hydroponic feed/fodder on wider scale, several issues
pertaining to HPFP technology, namely, economics, viability, sustainability and superiority of
hydroponic Fodder require further research on priority.
Result and Discussion
For sustainable dairy farming, quality green fodder should be fed regularly to dairy
animals. Hydroponic fodder is a good option in front of the farmer because it grows fast, it
contains a high nutrient value, and the most important thing is animals like to eat. Green fodder
is an important constituent in the feed of livestock. Due to many drastic Changes in agriculture

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system, animal rearing and increased population there is shortage of Green fodder to the
livestock. Green fodder production through hydroponics technology can be a real beneficial
alternative source to overcome the fodder deficiency in livestock sector with many advantage.
In developed countries where there is no dearth of quality feed and fodder, the hydroponic
production of fodder is less competitive than traditional fodder production when compared on
per kg dry matter basis.
References
1. Authored by Nisha Sharma, Somen Acharya, Kaushal Kumar, Narendra Singhand O.P. Chaurasia,
in the Journal of Soil and Water Conservation 17(4): 364-371,October-December 2018
2. Authored by Bikram Pradhan and Bandita Deo, published in Current Science, Vol.116, No. 5.
3. Authored by Shailesh Solanki, Nitish Gaurav, Geetha Bhawani and Abhinav Kumar, published in
International Journal of Advanced Research (IJAR), this paper focus on the challenges and
possibilities to bring soil less farming in India.
4. Authored by Awadhesh Kumar, published in Acta Scientific Agriculture (ISSN:2581-365X)
Volume 3, Issue 2, 2019.
5. Butler, J.D., 2006. Hydroponics as hobby growing plants without soil. Information Ofce of
University of Illinois 18(2), 11-32.
6. Choi, B., Lee, S.S., 2012. Effects of waste nutrient solution on growth of Chinese cabbage (Brassica
campestris L.) in Korea. Korean Journal of Environmental Agriculture 30(2), 125-131.
7. Folds, E., 2018. Where did hydroponics come from? Accessed on 16-08-2021.Liao, P., Liu, J., Sun,
L., Chang, H., 2020. Can the Adoption of Protected Cultivation Facilities Affect Farm
Sustainability? Sustainability 12(23), 70-99.
8. Miller, A., 2011. A Critical Appraisal of Current Development in Vertical Farming. Carleton
University publisher 8(15), 23-45.
9. Resh, H.M., 2013. Hydroponic Food Production: A Definitive Guidebook for the Advanced Home
Gardener and the Commercial Hydroponic Grower. CRC Publisher 11(23), 45-68.Resh, H.M.,
1998.
10. Hydroponics: Questions and answers for successful growing. Woodbridge Publisher 5(4), 12-
23.Yadav, D., 2020.

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4
Design and Synthesis of Heterocyclic Active Moieties of Pyrazoles and Its
Importance’s
Somwanshi A.R.
Department of chemistry, J.D. Patil Sangludkar Mahavidyalaya, Daryapur, Amravati (Maharashtra)India
(Corresponding author: Somwanshi A.R., Email: [email protected],Mob. No. 9822468944)

Abstract
Pyrazoles have totally concerned and great attention in organic and pharmaceutical fields, due
to their promising value as synthetic intermediates for the generation of various bioactive
compounds, Subsequently, the synthesis of pyrazoles is a significant focus on research for
synthetic organic chemists. Similarly, fused pyrazoles such as pyrazolo based pyridines and
pyrazolo pyrimidines have been broadly studied due to their various physiochemical and
biological utilities based on the important structural electronic properties of these N-
heterocyclic compounds. So, the synthesis of these fused heterocycles and of their derivatives
is of distinguished interest to both reveal novel derivatives and exposed the new application.
Several procedures have been explained in the literature for the preparation of pyrazoles and
of their fused systems in recent years, which mostly involve condensation reactions. And the
synthesized pyrazole-based derivatives were confirmed by 1H NMR and GCMS analysis.
Keywords: Pyrazole, N-Heterocyclic, bioactive, fused heterocycles
Introduction
Heteroaromatic compound pyrazole compound of 5-membered ring having two adjacent
nitrogen atoms [1]. NH-Pyrazoles are weak bases and also weak acids because they have
tendency to accept the protons (C=N) and also nitrogen atom (N-H) in pyrrole have tendency
to donate protons. [2] Similarly, the interaction between the heteroatoms and hydrogen i.e.,
hydrogen bonding is depending on structural units of pyrazoles [3].
A German chemist Ludwig Knorr in 1883, who tried to prepare the quinoline compound with
antipyretic activity [4]. But unfortunately synthesized the pyrazole instead to quinoline [5].
Knorr has first familiarized the of pyrazole to this heterocycles core to represent that it was
derived from pyrrole by the replacement of carbon atom by nitrogen [6]. the first to notice
antipyretic action of pyrazole-based compounds in man, which has encouraged the interest in
pyrazoles moiety [7]. Then in 1846, the Kosuge and Okeda extracted (a plant which is having
antimicrobial activity), 3-n-nonylpyrazole from Houttuynia cordata and also levo-β-(1-
pyrazolyl) alanine from watermelon seeds (Citrullus vulgaris) [8]. Until these findings it was
thought that pyrazoles could not be found naturally [9]. The adaptability of pyrazole moiety-
based compounds in biological utilities as well as synthetic also. It has been well recognized,
being even one of the furthermost studied compounds among the azole family, even though
there are so many natural products containing the pyrazoles moiety [10].
A N-heterocyclic compounds as like composed pyrazoles, substitutes pyrazoles are important
due to their wide applications [11]. Most of natural products shows the presence of fused
pyrazole structure. A compound pyrazolo[4,3-d] pyrimidine is naturally present in Formicin
A. Which is having different biological activity. Such as antiviral and antitumor.
In over-all, biological and pharmacological activities of these compounds containing pyrazole
core moiety fused with five- and six-membered heterocyclic compounds. In particular,
pyrazole-based pyridine and pyrimidines have taken part in drug discovery [12].

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EXPERIMENTAL SECTION
MATERIALS

Methanol (98%), Ethanol (98%) Acetonitrile (99%) and ethyl acetate (98%) were procured from Avra
chemical Pvt. Ltd. Sodium hydroxide (98%) zinc chloride Were acquired from Sisco Research
Laboratories Pvt. Ltd.
CHARACTERIZATION TECHNIQUE
The Chemical structure of synthesized compounds was confirmed by spectral data. 1H-NMR
spectra were recorded on BRUKER AVANCE NEO 500 MHz spectrometer using DMSO and
CDCl3 solvent and TMS as internal standards at SAIF, Punjab University, Chandigarh (India).
Chemical shifts are expressed in ppm. Mass spectrums were recorded on Thermo Scientific
TSQ 8000 Gas Chromatograph
General Reaction

Scheme 1. General Reaction for Synthesis of N-Heterocyclic Substituted Pyrazoles

Reactant (R1) (0.1 mmol) was heated with reactant (R2) (0.15 mmol) in oil bath at 100 ºC to
110 ºC, till complete removal of dehydrated water is ensured. The solid cream color precipitate
was obtained and it filtered, washed with methanol and recrystallized by ethanol with
preferable yield, m.p.-178ºC as cream- white colored crystalline solid. (Scheme 1.)
Table 1. Synthesis of N-heterocyclic Pyrazoles Derivatives
Sr. R1 R2 Product Time Yield
No in
Hrs.

1 4 86

2 4 81

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3 4 87

Table 2. Structural analysis of derivatives

Sr. Structure of Products Structural analysis by 1HNMR and GCMS
No

1 1
HNMR(500MHz,CDCl3):9.40(s,1H),6.747.20(m,4H),3.41(s
,3H),3.2(q,1H),2.0-2.16(t,2H),1.56-1.81(t,2H),1.6(t,1H)

GCMS: Cal m/z: 245.28, Found m/z: 244.16

2 1
HNMR(500MHz,CDCl3):16.4(s,1H),6.747.68(m,5H),3.41(s
,3H),3.2(d,1H),2.0-2.16(t,2H),1.56-1.81(t,2H),1.6(t,1H)

GCMS: Cal m/z: 273.11, Found m/z: 273.06

3 1
HNMR(500MHz,CDCl3):9.69(s,1H),7.68(s,1H),6.68(s,1H),
3.41(s,3H),3.2(q,1H),2.06-2.16(q,2H),1.56-
1.81(m,2H),1.6(t,1H)

GCMS: Cal m/z: 252.29, Found m/z: 251.40

Conclusion
Synthesis of pyrazole derivatives and even their various functional group substituents are well
established. Mostly, the bioactivity of the pyrazole derivatives was studied in detail. There are
recent attempts in understanding different properties of the pyrazole derivatives. In addition,
there are some challenges to overcome. These challenges include efficiency for high yields in
synthesis, generating novel pyrazole derivatives with bioactivity in the sub micromolar range,
and characterizing the properties of the derivatives accurately. For this reason, we believe there
is a need for investigating new synthetic routes, studying different properties and seeking new
applications of novel derivatives especially in blends with polymers. Therefore, the new trend
in pyrazole derivatives will be towards new applications in various areas.
Acknowledgement
The author is thankful to Department of chemistry, J.D. Patil Sangludkar Mahavidyalaya for
providing research facilities Authors are very much thankful to the Director, SAIF, Punjab
University Chandigarh for providing spectral data.
Conflict of Interest: None

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References
1) Bekhit, Adnan A., et al. "New heterocyclic hybrids of pyrazole and its bioisosteres: Design, synthesis
and biological evaluation as dual acting antimalarial-antileishmanial agents." European Journal of
Medicinal Chemistry 94 (2015): 30-44.
2) Farag, A. M., Ali, K. A., El-Debss, T. M., Mayhoub, A. S., Amr, A. G. E., Abdel-Hafez, N. A., &
Abdulla, M. M. (2010). Design, synthesis and structure–activity relationship study of novel pyrazole-
based heterocycles as potential antitumor agents. European journal of medicinal chemistry, 45(12),
5887-5898.
3) Abdelgawad, N., Ismail, M. F., Hekal, M. H., & Marzouk, M. I. (2019). Design, synthesis, and
evaluation of some novel heterocycles bearing pyrazole moiety as potential anticancer agents. Journal
of Heterocyclic Chemistry, 56(6), 1771-1779.
Fayed, E. A., Eissa, S. I., Bayoumi, A. H., Gohar, N. A., Mehany, A. B., & Ammar, Y. A. (2019).
Design, synthesis, cytotoxicity and molecular modeling studies of some novel fluorinated pyrazole-
based heterocycles as anticancer and apoptosis-inducing agents. Molecular diversity, 23, 165-181.
4) Nossier, E. S., Fahmy, H. H., Khalifa, N. M., El-Eraky, W. I., & Baset, M. A. (2017). Design and
synthesis of novel pyrazole-substituted different nitrogenous heterocyclic ring systems as potential anti-
inflammatory agents. Molecules, 22(4), 512.
Faisal, M., Saeed, A., Hussain, S., Dar, P., & Larik, F. A. (2019). Recent developments in synthetic
chemistry and biological activities of pyrazole derivatives. Journal of Chemical Sciences, 131, 1-30.
5) M Abdelrazek, F., M Gomha, S., H Abdelrahman, A., Metz, P., & A Sayed, M. (2017). A facile
synthesis and drug design of some new heterocyclic compounds incorporating pyridine moiety and their
antimicrobial evaluation. Letters in Drug Design & Discovery, 14(7), 752-762.
6) Gangurde, K. B., More, R. A., Adole, V. A., & Ghotekar, D. S. (2024). Design, synthesis and
biological evaluation of new series of benzotriazole-pyrazole clubbed thiazole hybrids as bioactive
heterocycles: Antibacterial, antifungal, antioxidant, cytotoxicity study. Journal of Molecular
Structure, 1299, 136760.
7) Reddy, G. M., Garcia, J. R., Yuvaraja, G., Venkata Subbaiah, M., & Wen, J. C. (2020). Design,
synthesis of tri‐substituted pyrazole derivatives as promising antimicrobial agents and investigation of
structure activity relationships. Journal of Heterocyclic Chemistry, 57(5), 2288-2296.
8)Castillo, J. C., & Portilla, J. (2018). Recent advances in the synthesis of new pyrazole
derivatives. Targets Heterocycl. Syst, 22, 194-223.
Karati, D., Mahadik, K. R., & Kumar, D. (2022). Pyrazole Scaffolds: Centrality in Anti-Inflammatory
and Antiviral Drug Design. Medicinal Chemistry, 18(10), 1060-1072.
9) Liu, X. R., Wu, H., He, Z. Y., Ma, Z. Q., Feng, J. T., & Zhang, X. (2014). Design, synthesis and
fungicidal activities of some novel pyrazole derivatives. Molecules, 19(9), 14036-14051.
10) Ansari, A., Ali, A., & Asif, M. (2017). Biologically active pyrazole derivatives. New Journal of
Chemistry, 41(1), 16-41.
11) Abd-El Gawad, N. M., Hassan, G. S., & Georgey, H. H. (2012). Design and synthesis of some
pyrazole derivatives of expected anti-inflammatory and analgesic activities. Medicinal chemistry
research, 21, 983-994.
12) Bakthavatchala Reddy, N., Zyryanov, G. V., Mallikarjuna Reddy, G., Balakrishna, A., Padmaja,
A., Padmavathi, V., ... & Sravya, G. (2019). Design and synthesis of some new benzimidazole
containing pyrazoles and pyrazolyl thiazoles as potential antimicrobial agents. Journal of Heterocyclic
Chemistry, 56(2), 589-596.

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QSAR analysis of sodium glucose co–transporter 1 (SGLT1) inhibitors

for anti-hyperglycaemic lead development
Archana Chapolikar1,*, Jyoti Dahatonde1, Pooja Gaikwad1, Ajaykumar Gandhi1
1
Department of Chemistry, Government College of Arts and Science, Chhatrapati Sambhajinagar,
Maharashtra, India, 431004 ([email protected] )
*Correspondence : [email protected], Mobile Number- (+91)9657538276.

Abstract: GA-MLR based QSAR evaluation has been effectuated on a small dataset of
SGLT1 inhibitors abide by OECD principles. Recent study revealed that the presence of o-
xylene moiety, hydrogen atoms exactly five bonds from hydrogen bond donor atoms and
ring carbon atoms within 2Å from hydrogen bond donor/acceptor atoms facilitate the SGLT1
inhibitory action of the compound. Developed QSAR models shown excellent performance
on all the statistical parameters with values well above the approved thresholds, such as, R2 =
0.79- 0.80, Q2LOO = 0.77, Q2LMO = 0.76, Q2Fn = 0.87–0.90, CCCext = 0.93-95. Applicability
domain
analysis and Y-randomization test also enhanced the credibility of this study.

Keywords: QSAR, SGLT1, anti-hyperglycaemics.

ABBREVIATIONS:

CADD- Computer Aided Drug Designing, SGLT1/2 - sodium glucose co–transporter

1/2, SMILES- Simplified Molecular-Input Line-Entry System, GA- Genetic Algorithm,
MLR- Multiple Linear Regression, QSAR- Quantitative Structure-Activity Relationship,
QSARINS- QSAR Insubria, OECD- Organization for Economic Co-operation and
Development, OFS- Objective Feature Selection, SFS- Subjective Feature Selection
1. INTRODUCTION
SGLT1 and SGLT2 play crucial role in renal glucose re-absorption. SGLT1 is present in
both the renal tubules and small intestine and are responsible for active glucose absorption.
SGLT1 is a low-capacity glucose transporter (relative to SGLT2), its higher glucose affinity
than SGLT2 and additional galactose transportation capability has own an identity of
anti-
hyperglycaemic target to SGLT1. Dual SGLT inhibitors i.e. inhibitors of both SGLT1 and
SGLT2 are under clinical trial and unfortunately no SGLT1 inhibitor based drugs in the
market are present.
C-phenyl 1-thio-D-glucitol scaffold based derivatives [10,11], Deuterated C‑Aryl
Glycoside[12], Indole‑N‑glucoside (TA-1887)[13], 1-methoxy-
6,8- dioxabicyclo[3.2.1]octane[14], benzocyclobutane-C-glycosides[15], 6-hydroxyl C-aryl
glucoside derivatives[16], benzocyclobutane-C-glycosides[17], 6-deoxy O-spiroketal C-
arylglucosides [18] are some classes of compounds tested for SGLT1 inhibitory potency.
But the search of sufficiently potent SGLT1 inhibitor with no or minimum side effect is still
on. To contribute to the development of better SGLT1 inhibitor L. Burggraaff did attempt to
identify SGLT1 inhibitors in silico (Burggraaff et al., 2019) with the application of
proteochemometrics through machine learning. There is still huge scope for the development

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

of SGLT1 inhibitors as possibly effective anti-hyperglycaemic agent.

2. MATERIALS AND METHODS

A. Selection of data-set:
For the present work, 107 C-aryl glucoside derivatives with in vitro SGLT1 inhibitory
activity in terms of half-maximal inhibitory concentration i.e. IC50 = 0.4 to 100000 nM were
used [19- 22]. The dataset consists of benzocyclobutane-C- (Placeholder1), deuterated
C‑Aryl Glycoside, series of C-phenyl D-glucitol derivatives, 6-deoxy O-spiroketal-C-
arylglucosides, C-phenyl 1-thio-D-glucitol, and C5-fluoro-hexose derivatives. A dataset
truly comprised of diverse set of molecules with plenty of pharmacophoric features
embedded along with the different scaffolds and hence has covered large chemical space. A
QSAR model developed using such a dataset will certainly have large applicability domain
that cover the class of the compounds presently under the process of optimization towards
more potent SGLT1 inhibitors to treat hyperglycaemia.
The IC50 values in nanomolar (nM) unit were converted into molar (M) unit by multiplying
factor 10−9. The five least and the five most active C-aryl glucoside derivatives from the
present dataset are depicted in Figure 1 to demonstrate the variation in bio-activity with
chemical features. The SMILES strings with the reported IC50 (in nM and M unit) and pIC50
values for all the molecules are given in Supplimentary information. Before QSAR analysis,
the IC50 values in the molar unit were converted into pIC50 by using formula 𝑝𝐼𝐶50 = − log10
𝐼𝐶50 for ease of the data handling.

Figure 1: Variations in activity and chemical structure in the dataset used for development of QSAR Models

B. Molecular Structure Drawing and Optimization:

Free and open source software, ChemSketch 12 Freeware (www.acdlabs.com) is
used to draw 2D structures and further conversion to 3D structures is achieved using
OpenBabel
2.4.0. A geometry optimized molecule is characterized by its lowest energy i.e. most stable
conformation. Geometry optimization ensures the normalization of all such a physico-
chemical properties for all the molecules from dataset and makes it a necessarly important
step prior to molecular descriptor calculation. As a step forward, force field MMFF94
available in TINKER is used for optimization of the molecules.
C. Molecular Descriptor Calculation and Objective Feature Selection (OFS):
A molecular descriptor is a structural and physico-chemical property of a molecule
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or specific part of the molecule. More than 18,000 molecular descriptors were calculated for
each molecule using PyDescriptor and PaDEL. In the data pruining step, Objective Feature
Selection (OFS) in QSARINS v2.2.4 screened out multi-collinear and spurious molecular
descriptors (i.e. with nearly constant values >95%, co-linearity |R| >0.95) and contracted
molecular descriptor pool with 1145 variables is generated. Contracted though, molecular
descriptor pool has covered adequately comprehensive chemical space being comprised of
0D– to 3D– descriptors, structural, constutional properties and charge descriptors etc.
D. Subjective Feature Selection, QSAR Model – Development and Validation:
In accordance with the OECD principles, firstly, apt variable selection method GA– MLR
in Subjective Feature Selection (SFS) operation in QSARINS v2.2.4 is used to perform
simple and easy to interpret QSAR models. Then, all the derived models were subjected to
the thorough statistical validation, Y-scrambling and Applicability Domain analysis. The
steps in QSAR building process:
I. Random split operation in QSARINS v2.2.4 split given dataset into a training set with
35 molecules (80%) and a prediction set with 9 molecules (20%) prediction. Molecules in
training set were utilized for QSAR model development and external validation was
performed on 9 molecules in prediction set.
II. QSAR models were built using Subjective Feature Selection (SFS) operation in
QSARINS v2.2.4 (at default settings) by setting Q2LOO as fitness function. Insignificant
increase in Q2LOO value was observed after 5 variables and hence to avoid overfitting SFS
opertion is confined to 5 variables which additionally helped in deriving easy and informative
QSAR models. (See supplementary information Table S2)
III. (a) Leave-One-Out (LOO) and Leave-Many-Out (LMO) parameter based internal
validation; (b) External validation; (c) Y-scrambling and model Applicability Domain (AD)
analysis, performed for legitimate validation.
IV. Performance of each model, measured by close inspection of the various statistical
parameters meter the robustness of the GA-MLR based QSAR model. The QSAR model
with best values of these parameters and with best predicative ability is selected.

3. RESULTS

Statistical QSAR
After removing all the insignificants terms, the following GAMLR QSAR model-1 and
model- 2 were obtained from GA-MLR analysis.
GAMLR QSAR Model-1 (Training Set: 80% and Prediction Set: 20%)
𝑝𝐼𝐶50 = 22.737(±0.2.253) + 0.608(±0.275)_𝒓𝒊𝒏𝒈𝑪_𝟐𝑨
+ 0.289(±0.121)𝒐𝒏𝑯𝟓𝑩 − 0.466(±0.173)𝒓𝒊𝒏𝒈𝑺_𝑪_𝟖𝑨𝒄
− 634.57(±82.98)𝑷. 𝟕
GAMLR QSAR Model-2 (Training Set: 80% and Prediction Set: 20%)

𝑝𝐼𝐶50 = 18.274(±0.2.440) + 0.657(±0.308)𝑲𝑹𝑭𝑷𝟑𝟓𝟗𝟔

+ 0.258(±0.116)𝒇𝒅𝒐𝒏𝑯𝟓𝑩
− 0.206(±0.073)𝒇𝒓𝒊𝒏𝒈𝑺𝑯𝟔𝑩
− 424.535(±82.051)𝑷. 𝟕 (1)

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Statistical The statistical parameters for developed

Values
Parameters GAMLR QSAR models 1 and 2 have been
Model Model 5.2 presented in Table 1. A good number of
5.1 statistical parameters for model 1 and 2, which
FITTING are related to fitting, internal and external
𝑅2 0.79 0.80 validation and Y-scrambling, have been
𝑅2 0.78 0.79 calculated. From Table 1, it is clear that R2TR,
𝑎𝑑𝑗. CCCTR, CCCcv, R2adj, and F satisfy the
𝑅2 − 𝑅2 0.01 0.01 recommended threshold value, which shows that
𝑎𝑑𝑗.
𝐿𝑂𝐹 0.32 0.32 the QSAR models are statistically robust with
𝐾𝑥𝑥 0.22 0.23 adequate number of molecular descriptors in the
∆𝐾 0.14 0.15 models. Although the data are obtained from
different publications, they pass the statistical
𝑅𝑀𝑆𝐸𝑇𝑅 0.52 0.51
compatibility as the values of statistical
𝑀𝐴𝐸𝑇𝑅 0.40 0.39
parameters for fitting criteria such as R2adj =
𝑅𝑆𝑆𝑇𝑅 22.85 22.48
0.78, R2-R2adj = 0.01, LOF = 0.32, CCCTR =
𝐶𝐶𝐶𝑇𝑅 0.88 0.89
0.88, etc. are well above the approved threshold
𝑠 0.53 0.53
values for both models. From Fitting parameters
𝐹 77.42 79.01 such as, R2TR, CCCTR, CCCcv, R2adj, and F satisfy
INTERNAL
VALIDATION
the recommended threshold value which shows
0.77 0.77 that the QSAR models are statistically robust
𝑄2 with adequate number of molecular descriptors
𝐿𝑂𝑂
𝑅2 − 𝑄2 0.02 0.03 in them. The values for different cross-
𝐿𝑂𝑂 validation parameters such as Q2LOO, RMSEcv,
𝑅𝑀𝑆𝐸𝐶𝑉 0.55 0.54 MAEcv, CCCcv, and Q2LMO support the statistical
𝑀𝐴𝐸𝐶𝑉 0.42 0.41 robustness of the QSAR models. The graphs;
𝑃𝑅𝐸𝑆𝑆𝐶𝑉 25.60 25.53 Experimental endpoint against Predicted
𝐶𝐶𝐶𝐶𝑉 0.87 0.87 endpoint for both the models [Figure 4a & 5a]
𝑄2 0.76 0.76 and experimental endpoint against residuals
𝐿𝑀𝑂 [Figure 4b & 5b] represent the performance of
𝑅2 0.05 0.05 the QSAR model 1 and 2, respectively, on both
𝑌𝑆𝐶𝑅 Training and Prediction set molecules.
𝑅𝑀𝑆𝐸 𝐴𝑉𝑌𝑆𝐶𝑅 1.10 1.10
𝑄2 -0.07 -0.07
𝑌𝑆𝐶𝑅
The external predictive ability of the models is
established by the high values of R2 EXT, Q2 F1,
EXTERNAL Q2F2, Q2F3, and CCCEXT. In short, the developed
VALIDATION QSAR models fulfill the recommended threshold
𝑅𝑀𝑆𝐸𝐸𝑋𝑇 0.36 0.40 values for many internal and external validation
𝑀𝐴𝐸𝐸𝑋𝑇 0.32 0.35 parameters. In addition, for a better validation of
𝑃𝑅𝐸𝑆𝑆𝐸𝑋𝑇 2.78 3.43 derived models, the model applicability domain
2
𝑅𝐸𝑋𝑇 0.90 0.88 (AD) was assured by plotting Williams plots for
𝑄𝐹21 0.90 0.87 models 1 [Figure 4b & 5b]. Therefore, these
𝑄𝐹22 0.90 0.87 models are statistically robust and possess good
𝑄𝐹23 0.90 0.87 external predictive ability. Moreover, fulfilment
𝐶𝐶𝐶𝐸𝑋𝑇 0.95 0.93 of recommended threshold values for many
𝑟𝑚2 𝑎𝑣𝑔. 0.85 0.76 parameters as well as low R2 value after Y-
𝑟𝑚2 𝑑𝑒𝑙𝑡𝑎 0.03 0.13 randomization indicate that the model is not
𝑘′ 1.00 0.99 developed by chance.
The performance of the QSAR Model 1 and 2 for
𝑘 1.00 1.00

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each molecule from used dataset is given in Supplimentary Information respectively.

Compound 29 (Exp. pIC50 =7.036 Pred. pIC50 = 7.042, PMER value = 0.006, HAT i/i= 0.037)
and 85 (Exp. pIC50 =7.456, Pred. pIC50 = 7.457,
PMER value = 0.001, HAT i/i= 0.1 10) are the best predicted for their pIC50 values by model
1 and 2, respectively. Both compound 29 and 85 fall well within the AD of respective models
as reflected from HAT i/I values. Possible reason for the best bioactivity P for the compound
29 [Figure 2] might be the presence of simple structural features, molecular planarity in a side
chain of the glucoside scaffold maintained by simple methyl substituent and a fused ring
substitution (benzocyclobutane) that minimized the errors in the bioactivity Prediction due to
conformational changes in the molecules. Compound 85 (Figure 3) is with more number of
pharmacophoric feature with large number of hydrogen bond donor and acceptor atoms (O, N,
etc.). This might have helped molecule to attain optimum value for the molecular descriptors
wherefrom QSAR equation is developed in subjective feature selection.

Figure 2: Representation of the best predicted compound by QSAR Model 1 and 2

Collective result of all these causes could be the reason for best SGLT1 inhibitory activity
prediction for these compounds. Models developed in present study failed to predict precise
bioactivity for some of the compound (see Figure 3). Compound 102 is the worst predicted
compound (Exp. pIC50 =4.449 Pred. pIC50 = 5.784, PMER value = 1.335, HAT i/i= 0.038)
for its SGLT1 inhibitory activity by model 1 whereas compound 45 is the worst predicted
compound (Exp. pIC50 =5.933, Pred. pIC50 = 7.348, PMER value = 1.415, HAT i/i= 0.024)
for SGLT1 inhibitory activity by model 5.2. In both the models, compound 14 is the third
worst predicted compound (Exp. pIC50 =7.578, by model 5.1; Pred. pIC50 = 8.893, PMER
value
=1.306, HAT i/i= 0.079 and by model 5.2; Pred. pIC50 = 8.858, PMER value =1.271, HAT
i/i= 0.079).

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Figure 3: Representation of worst predicted and outlier compounds

Figure 4: (a) Graph of Experimental vs. Predicted Endpoint values (b) Williams plot for QSAR Model-1

Figure 5: (a) Graph of Experimental vs. Predicted Endpoint values (b) Williams plot for QSAR Model-2

Descriptive QSAR fdonH5B

The molecular descriptor fdonH5B encode the information on frequency or number of
Hydrogen atoms exactly five bonds from HBD atom. It has positive coefficient, thus increase
in the value of fdonH5B increase the pIC50 value i.e. SGLT1 inhibitory potency of the
compound. To support this observation, a pair of compound 44 (fdonH5B = 2; pIC50 = 6.754)
and 30 (fdonH5B = 1; pIC50 = 6.686) is appropriate. Compound 17 (fdonH5B = 0; pIC50 =
8.301) and 16 (fdonH5B = 3; pIC50 = 8.398) is yet another pair of compound that appropriately
support the same observation (Figure 6). The substituent –Cl in compound 17 on replacement
by –CH3 added three Hydrogens which got placed exactly five bond from HBD i.e. –OH group
ortho glycoside residue.

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ringS_C_8Ac Figure 6: Illustration of the molecular descriptor fdonH5B

The molecular descriptor ringS_C_8Ac encode the information on the sum of partial charges
of ring Sulfur atoms within 8Å from C atom. As it has negative coefficient, decrease in the
value of ringS_C_8Ac enhance the SGLT1 inhibitory potency of the compound. The
significance of this molecular descriptor is highlighted by comparing compound 9
(ringS_C_8Ac = 2.337; pIC50 = 8.194) with 10 (ringS_C_8Ac = 1.897; pIC50 = 8.328) wherein
decrease in the value of ringS_C_8Ac furnished more potent SGLT1 inhibitor (see Figure
7). The close observation of the structures of compound 9 and 10 reveal that this molecular
descriptor indirectly provide information on substituency on ring containing Sulfur atom. In
compound 9, the electron donating inductive effect of –CH2- group is limited by an electron
withdrawing inductive effect of ester –C(O)O group whereas in compound there is ether
functionality that too separated by one more CH2 group and hence an electron donating effect
of CH2 group is retained. This feature in compound 10 diminish the partial positive charge
on ring Sulfur.

da_ringC_2A

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Figure 7: Illustration of the molecular descriptor ringS_C_8Ac

The molecular descriptor, da_ringC_2A encode the information on number of ring Carbon
atoms within 2Å from HBD and HBA. It has positive coefficient and hence increase in the
value of this descriptor cause increase in the SGLT1 inhibition activity of the compound.
Comparison of compound 44 (da_ringC_2A =2; pIC50 = 6.754) with 30 (da_ringC_2A
=1;
pIC50 = 6.686) rationalize the observation. Compound 48 (da_ringC_2A =3; pIC50 = 7.444)
with 47 (da_ringC_2A =4; pIC50 = 7.638) is yet another pair of the compound to support this
fact.
KRFP3596
It is a Klekota-Roth fingerprint abbreviated as KRFP. It encode the information on the
presence of various chemical substructures. KRFP3596 molecular descriptor encode
specifically the presence of substructure represented by o-xylene moiety (see Figure 8). This
molecular descriptor has positive coefficient and hence the presence of o-xylene substructure
own better SGLT1 inhibitory potency to the compound. This observation can be supported
by the observation of whole dataset. All the most active compounds with pIC50 ≥ 7.036. o-
xylene substructure is present (KRFP3596 = 1) whereas in the least active compounds with
pIC50 ≤
5.514 (with few exceptions) there is no o-xylene substructure present (i.e. KRFP3596 = 0).
In Figure 8 o-xylene moiety quantitatively encoded as KRFP3596 is depicted using
compound 4 (a most active SGLT1 inhibitor of the series).

ASP.7 Figure 8: Illustration of the molecular descriptor KRFP3596

ASP.7 i.e. average simple path, order 7 abbreviated as ASP.7 is one of the χ Chi Path descriptor.
It has negative coefficient in both the models. Therefore, decrease in the value of ASP.7
increase the value of pIC50 i.e. SGLT1 inhibitory potency of the compound. Significance of
the smaller value of ASP.7 molecular descriptor for better SGLT1 inhibitory potency is
supported
by comparing compound 1 (ASP.7 = 0.027, pIC50 = 7.652) with compound 2 (ASP.7 = 0.026,
pIC50 = 9.398). Decrease in the value of ASP.7 by just 0.001 unit elevated the potency of
compound 3 by about 1.7 unit.
fringSH6B

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The molecular descriptor, fringSH6B encode the information on the number of Hydrogen
within 6 bonds from ring Sulfur. It has negative coefficient and hence decrease in the value
of fringSH6B could increase the SGLT1 inhibitory potency of the compound. The least
active SGLT1 inhibitor of the series i.e. compound 96 there are 6 such a hydrogens are
present (Figure 9) and last 7 least active compounds have at least 2 such hydrogens (i.e.
fringSH6B =2 in their structure. On the contrary, the 4 most active compounds of the series
have only 1 or even no (0) such a hydrogens (Figure 9). This molecular descriptor provide an
easy ground for optimization of present SGLT1 inhibitors toward more potent leads. For
example, in compound 4 (the most potent SGLT1 of the series) just repositioning of –Cl that
replace hydrogen at 6 bond from sulfur could furnish more potent SGLT1 inhibitor. In case
of presence of -OCH3 group as a bearer of such hydrogens can be replaced by –OCF3 or –OH
group and more SGLT1 inhibitor can be expected.
Figure 9: Illustration of the molecular descriptor fringSH6B

4. CONCLUSIONS
Statistical accomplishment of both the models is quite satisfactory as reflected from values
of the various statistical parameters that are well above the approve threshold values. High
value of Q2LOO = 0.77, Q2LMO = 0.76, R2EXT = 0.88-0.90, and Q2Fn = 0.87-0.90 and

CCCEXT
= 0.93-0.95 mark the acceptable predictive ability of the Model 1 and 2. Applicability domain
analysis based on leverage approach provided an information on the most appropriate classes
of the compounds with distinguished scaffold and pharmacophoric features for reliable
prediction of SGLT1 inhibition potential prior to their wet-lab synthesis and/or in-vitro/in-
vivo
studies. Information on PMER values for all the compounds and best predicted compound(s)
help to have precise guess on the predicted activity. The knowledge of worst predicted
compound(s) and/or outlier(s) help the user to identify compound(s) unsuitable for the
application of the models for reliable SGLT1 inhibitory potential.
Presence of o-xylene moiety, Hydrogen atoms exactly five bonds from HBD atom and ring
Carbon atoms within 2Å from HBA and HBD atoms, facilitate the SGLT1 inhibitory action
of the compound, as suggested by QSAR evaluations. In the course of optimization of the
existing SGLT1 inhibitors within applicability domain of the models developed, it is advisable
to ensure the more number of Hydrogens exactly five bond and ring Carbons within 2Å from
HBA and/or HBD atoms. Introduction of ring substituent within 2Å from HBA and HBD
atoms or substituting existing ring system with optimum number of functionalities with HBA
and HBD atoms could suffice to have required structural modification and hence highly
recommended during structural derivatization. QSAR evaluation also suggest that the ring

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Sulfur atoms within 8Å from Carbon atom, Hydrogen atoms within 6 bonds from ring Sulfur
are detrimental to the activity of the SGLT1 inhibitors. In the compounds from dataset, sulfur
present in two different scaffold systems viz. S-glycoside ring and thiophene ring. It is
observed that, occurrence of S-glycoside moiety set to zero towards the end of most potent
SGLT1 inhibitors. However, thiophene has not proved that detrimental to SGLT1 inhibitory
potency of the compound as it is present in some of the relatively more potent SGLT1
inhibitors. Removal of thiophene is not advisable but, it is suggestive to not to substitute
thiophene ring with substituent that could intensify the positive charge on Sulfur of the
thiophene through electron withdrawing inductive/resonance effect such as –Cl, -COR, -
C(O)OR etc.
In conclusion, the present QSAR analysis was effective in identifying interdependent,
interconnected astounding structural features which are otherwise hard to recognize by mere
observation. Strategic optimization of the present SGLT1 inhibitors within applicability
domain of the model with recommended Structural modifications could lead to the discovery
of more active SGLT1 inhibitor(s) as an effective anti-hyperglycaemic agent with acceptable
ADMET profile.
ACKNOWLEDGMENTS: The authors are thankful to Dr. Paola Gramatica and her team
for providing QSARINS-v2.2.4 and developers of TINKER, ChemSketch 12 Freeware
(ACD labs), and PyDescriptor for providing the free versions of their software.
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type 2 diabetes treatment, Bioorganic Med. Chem. 27 (2019), pp. 394–409.
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absorbable sodium-dependent glucose cotransporter 1 (SGLT1) inhibitor (TP0438836) for the
treatment of type 2 diabetes, Bioorganic Med. Chem. Lett. 28 (2018), pp. 3534–3539.
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of benzocyclobutane-C-glycosides as potent and orally active SGLT1/SGLT2 dual inhibitors,
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[33] J. Huang and X. Fan, Why QSAR fails: An empirical evaluation using conventional computational
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6
Preparation and Acetone Gas Sensing Properties of Cd Substituted
Magnesium Ferrites
A. V. Kadu
Prof Ram Meghe College of Engineering and Management, Badnera, Amravati, M.S. India.
Mail id: [email protected]

Abstract :
In this work, we report the preparation and gas sensing performance of pure and doped
MgFe2O4 were prepared by sol gel method. The structural characteristics of the material were studied
by using X-ray diffraction and Scanning Electron Microscopy. The XRD pattern shows a
nanocrystalline solid solution of MgFe2O4 with an orthorhombic phase and the crystallite sizes is found
to be in the range of 30-40 nm. The gas sensing performance of the unmodified and surface modified
films was tested for various gases such as H2S, NH3, LPG and acetone. Mg0.4Cd0.6Fe2O4 powder showed
large response to 200 ppm acetone gas at an operating temperature 1800C. The sensitivity, selectivity
of Mg0.4Cd0.6Fe2O4 thick films was measured.
Keywords: Acetone sensor, Mg0.4Cd0.6Fe2O4, Gas sensing properties, Response time.
1. Introduction:

Magnesium ferrites (MgFe2O4) have emerged as promising candidates for gas sensing applications
owing to their inherent chemical stability, high surface area, and tunable electrical properties. The
ability to detect volatile organic compounds (VOCs) such as acetone is of particular interest due to its
relevance in various industrial, environmental, and healthcare sectors. However, enhancing the
sensitivity, selectivity, and response time of gas sensors based on magnesium ferrites remains a key
challenge.
Dopant-mediated modifications offer a promising strategy for tailoring the gas sensing properties of
magnesium ferrites. Among the dopants explored for this purpose, cadmium (Cd) substitution has
garnered significant attention due to its potential to influence the structural and electronic properties of
the host material. By introducing Cd ions into the magnesium ferrite lattice, it is possible to induce
changes in the surface morphology, defect structure, and gas interaction mechanisms, thereby
enhancing the sensing performance towards acetone and other VOCs.
The detection of acetone gas holds immense importance in various applications, including industrial
process monitoring, environmental pollution control, and medical diagnostics. Reliable and efficient
gas sensors capable of detecting acetone at low concentrations are essential for ensuring workplace
safety, environmental compliance, and early disease diagnosis.
Chemical methods [1-3], microwave sintering method [4,5], Citrate precursor method [6], wet chemical
method [7] and Sol-gel method [8] etc. The sol-gel method has been identified as the most
straightforward approach for preparing ferrites in bulk form. In this paper the results are presented of
the ferrites prepared by using sol gel method.
The development of Cd-substituted magnesium ferrite-based gas sensors presents an exciting
opportunity to address these challenges and advance the state-of-the-art in gas sensing technology.
Structural properties of Mg2+ and Al3+ co-substituted lithium ferrites were studied by Modi et al [9]. X-
ray and infrared studies of chromium substituted magnesium ferrite was reported by Kawade et al [10]
and found that, the lattice parameters decrease with Cd3+ substitution and the X-ray density decreases
as Cd3+ content increases. The distance between magnetic ions in both octahedral and tetrahedral sites
decreases with increase in Cd3+.
In this research paper, we present a systematic investigation of the preparation and acetone gas sensing
properties of Cd-substituted magnesium ferrites. Through a combination of synthesis techniques,
structural characterization, and gas sensing measurements, we aim to elucidate the influence of Cd
doping on the microstructure, surface chemistry, and sensing performance of the synthesized materials.

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2. Experimental Details
2.1. Material Synthesis
Analytical grade reagents were utilized throughout the experiment. Nanostructured MgFe 2O4
and Mg0.4Cd0.6Fe2O4 were synthesized by the sol-gel method. Initially, a precursor was prepared using
the sol-gel citrate method, employing the stoichiometric ratio of ferrous nitrate, magnesium nitrate,
cadmium nitrate, and citric acid. This precursor was then dissolved in ion-free water at 80°C for 2 hours.
Subsequently, ethylene glycol was introduced with continuous stirring to achieve a homogeneous and
stable sol. The solution underwent further heating in a pressure vessel at approximately 130°C for 12
hours. Throughout this process, the transparent solution underwent a transformation into a gel state with
notably high viscosity. The material was subsequently subjected to heating in a furnace at 350°C for 3
hours, resulting in a vigorous combustion that spontaneously propagated until all the gel was consumed,
leaving behind a loose powder. This powder is calcination at 650°C for 6 hours to enhance the
crystallinity of the materials.
2.2. Characterization techniques
The synthesized samples were characterized for their structure and morphology by X -ray
powder diffraction (XRD; Siemens D5000) and Scanning electron microscopy (SEM; Hitachi-800).
The X-ray diffraction data were recorded by using CuKα radiation (1.5406 A0).
2.3. Measurement of sensing properties

The gas-sensing properties of prepared Mg0.4Cd0.6Fe2O4 powders were studied for reducing
gases such as H2S, NH3, LPG and acetone whose concentration were fixed at 1000 ppm in air. The gas
sensitivity (S) was defined as: S = (Ra −Rg)/Ra =ΔR/Ra; where, Ra and Rg are the resistance of sensor
in air and the test gas, respectively. The gas-sensing properties were measured in a temperature range
of 50 – 3500C.
3. Results and Discussion
3.1. X-ray Diffraction Study

The XRD pattern of the Mg0.4Cd0.6Fe2O4 prepared by sol-gel method calcined at 6500C is presented in
Figure 1. Study of X-ray diffraction reveals that all the compositions under investigations were found
to be face centered cubic spinel structure. The XRD pattern indicates that the product has high degree
of crystallinity judged from the high and sharp diffraction peaks. The average particle size of the
nanocrystalline Mg0.4Cd0.6Fe2O4 according to the scherrer formula were in the range of 30–40 nm.

Fig 1 XRD pattern of Mg0.4Cd0.6Fe2O4 calcined at 6500C

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3.2 Scanning Electron Microscopy

The powder sample's morphology was examined using scanning electron microscopy (SEM).
Figure 2 displays the SEM image of the Mg0.4Cd0.6Fe2O4 powder, revealing a uniform distribution of
grain sizes with a slight tendency for agglomerate formation. The formation of polycrystalline material
led to particle sizes ranging between 30 to 40 nm.

Fig 2 SEM image of Mg0.4Cd0.6Fe2O4 calcined at 6500C

3.3. Gas Sensing Properties
Figure 3 illustrates the sensor response (S) plotted against the operating temperature for
undoped MgFe2O4 nanopowder, which was calcined at 650°C for 6 hours, when exposed to different
reducing gases such as H2S, NH3, LPG, and acetone. The sensor exhibits a notably higher response to
acetone gas compared to LPG, NH3, and H2S at an operating temperature of 250°C.

To enhance the gas response properties, various atoms or additives are introduced into the base
sensing semiconductor. Figure 4 illustrates the gas response concerning different amounts of Cd-doped
MgFe2O4 (x = 0.2, 0.4, 0.6, and 0.8). The response to acetone gas exhibits a consistent increase with the
rise in Cd concentration. The highest response is observed for Mg0.4Cd0.6Fe2O4 (x = 0.6) due to the
increased availability of sites for oxygen adsorption, facilitating the oxidation of the test gas.
Conversely, a decrease in response may stem from an inadequate number of available sites on the
surface. The partial replacement of Mg by Cd ions leads to a reduction in grain size, resulting in a higher
density of grain boundaries and consequently, an increased effective exposure area of the film to
ammonia gas. Additionally, the chemical composition of the semiconductor is a critical parameter
influencing its sensing performance. Indeed, the composition alone can impact the microstructure and
thereby dictate the sensing properties.

Fig 3 Gas sensing characteristics of undoped MgFe2O4 for various reducing

gases as a function of operating temperature.

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Fig 4 Sensor Response of MgFe2O4 doped with different amount of Cd

calcined at 6500C. (a) x=0.2, (b) x=0.4, (c) x=0.6and (d) 0.8

Figure 5 depicts the gas response of Mg0.4Cd0.6Fe2O4 across various operating temperatures. It
is evident that the sensor exhibits a notably higher response to acetone gas compared to H2S, NH3, and
LPG at 180°C. This heightened sensitivity to acetone gas can be attributed to the surface modification
achieved by Cd over the Mg0.4Cd0.6Fe2O4 film. The enhanced selectivity can be elucidated as follows:
Upon adsorption of O2- on the surface of Mg0.4Cd0.6Fe2O4, it captures electron(s) from the n-type
semiconductive Mg0.4Cd0.6Fe2O4 body due to the strong electronegativity of the oxygen atom, resulting
in the formation of negatively charged chemisorbed oxygen species such as O2−, O−, and O2−.
Consequently, the electron concentration within the n-type Mg0.4Cd0.6Fe2O4 diminishes, thereby leading
to an increase in the material's resistance.

Figure 6 shows the dependence of gas response of the Mg0.4Cd0.6Fe2O4 sensor on the
concentration level of acetone gas at 180oC. It is clear from the graph that with the increase in the
concentration, the response increases linearly up to 200 ppm of acetone gas, after that it saturates. The
graph also indicates that at low concentration response has a linear relationship with concentration
because there may be sufficient number of available surface states to act on acetone gas. After 200 ppm
level of acetone gas, the curve flattens because there would not be enough ionosorbed oxygen species
to contribute to detecting mechanisms.

Fig 5 Response to different reducing gases of Mg0.4Cd0.6Fe2O4 as a function of operating

temperature.

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

1.0

0.8

Response
0.6

0.4

0.2
50 100 150 200 250 300
Gas Concentration

Fig 6 Response of Mg0.4Cd0.6Fe2O4 to acetone gas of different

concentration at an operating temperature 180oC.

Figure 7 shows the response time of the sensor at 180oC. The response time in this case is ~ 40
sec. It was observed that the response time comes to saturated at 55 sec. It suggests that after this time
there is no more O– species left to react with the acetone gas vapour. It also indicates that by increasing
the surface area i.e. increasing the grain size of the film, one can increase the response time.

1.0

0.9

0.8

0.7
Response

0.6

0.5

0.4

0.3

0.2
0 10 20 30 40 50 60 70 80 90 100 110
Response Time (Second)

Fig 7 Response characteristics of Mg0.4Cd0.6Fe2O4 at 180oC.

4. Conclusion

In this study the preparation and gas sensing properties of both pure and doped MgFe 2O4 materials
synthesized through a chemical method. Structural analysis via X-ray diffraction and scanning electron
microscopy revealed a nanocrystalline solid solution of MgFe2O4 exhibiting an orthorhombic phase,
with crystallite sizes ranging between 30-40 nm. Gas sensing experiments were conducted on
unmodified and surface-modified films, assessing their responses to various gases including H2S, NH3,
LPG, and acetone. Specifically, Mg0.4Cd0.6Fe2O4 powder demonstrated a significant response to 200
ppm acetone gas at an operational temperature of 180°C. The sensitivity and selectivity of
Mg0.4Cd0.6Fe2O4 thick films were also evaluated. These findings underscore the potential of MgFe2O4-

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

based materials for gas sensing applications, particularly in detecting acetone gas, with implications for
diverse industrial and environmental monitoring scenarios.

References :

[1] I.Z. Rahman and T.F. Ahmed., J. Mag. and Mag. Mater., 2005, 290,1576.
[2] E.E. Sileo and S.E. Jacobo., Phys.B: Condens. Matter., 2004, 354,241.
[3] M.V. Islam, T. Abbas, S.B. Niaz. Z. Ahed, S. Sabeen and M.A. Choudary., Solid State
Comm., 2004, 130, 353.
[4] A. Bhaskar, B. Rajinikanth and S.R. Murthy., J. Mag. and Mag. Mater. , 2004, 283,109.
[5] Y.J.Yang, C.I Sheu, S.Y. Cheng and H. Chang., J. Mag. and Mag. Mater., 2004, 284, 220.
[6] A.Thakur and M. Singh., Ceramics Inter., 2003, 29, 505.
[7] K.M. Jadhav, V.B. Kawade, K.B. Modi, G.K. Bichile, R.G. Kulkarni., Phys.B: Conds.
Matter. 2000, 291, 379.
[8] S. Yan, J. Geng, L. Yin and E. Zheu., J. Mag. and Mag. Mater., 2004, 277, 84.
[9] K. B. Modi J. D. Gajera, M. C. Chhantbar, K. G. Saija, G. J. Baldha and H. Joshi., Mater.
Lettrs., 2003, 57,4049.
[10] V. B. Kawade, G. K. Bichile and K. M. Jadhav., Mater. Lettrs.,2000, 42, 33.

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

Studies of Acoustic Properties of Substituted Chalcone in Different

Percentage of Dioxane-Water Mixture

A. D. Khambre*
*Departments of Chemistry, Sant Gadge Baba Amravati University, Amravati (MS), India
Sant Gadge Baba Amravati University,Amravati
Email id –[email protected]

ABSTRACT--Ultrasonic velocity and density measurement of synthesized Dibromo chalcone

ligand were carried out in different percentage of Dioxane-water mixture at 30 ± 0.10C. The
obtained experimental data have been used to evaluate acoustical parameters such as
intermolecular free length, specific acoustic impedance, relative association, adiabatic
compressibility, apparent molal compressibility, and apparent molal volume. These parameters
are used to interpret solute-solvent, solute-solute interaction in the system.
Keywords --- 1,4-Dioxane, ligand, Ultrasonic velocity, intermolecular free length,metal ions
etc.
INTRODUCTION
The ultrasonic velocity technique is one of the most powerful techniques for studying
the nature of intermolecular interaction in liquid. The ultrasonic velocity and other acoustic
parameters can be measured with great accuracy and provides a powerful way to determine
intermolecular interactions. The ultrasonic studies are useful in extensive research in different
field of science [1-4]. This is because of its ability of characterizing physicochemical behaviour
of liquid medium[5-6]. The measurements of ultrasonic velocity are helpful to interpreted
solute- solvent, ion-solvent interaction in aqueous and non-aqueous medium[7-8].Numerous
workers have done the acoustical study by the measurement of density and ultrasonic velocity
of different aqueous and non-aqueous system at different temperature, different concentration
of solute and in different percentage of organic solvent[9-11].The ultrasonic and viscometric
studies of œ amino acids in different percentage of aqueous dioxane systems have been carried
out by Raut et al[12]. The ultrasonic interferometric investigations of 3-(chloroaryl)-5-aryl-1-
substituted pyrazolines in different percentage of dioxane medium at different temperature
have studied by Deshmukh and Raghuwanshi[13]. They are reported that the ultrasonic
velocity decreases with increase in percentage of organic solvent.Thorat S. A. and Thakur S.
D.have studied the ultrasonic behaviour and molecular interaction of substituted 3,5-diaryl
isoxazoline in different percentage DMF-water mixture at 305 K. They reported that there is
weak solute-solvent interaction in all systems[14]. Deosarkar et al investigated the acoustical
studies of some pyrazoles in different percentage of dioxane-water mixture at 303.15 k at 2
MHz frequency. They observed that the interactions are exist between pyrazoles and dioxane
water mixture and solute-solvent interactions are more favorable than other interactions [15].
The ultrasonic behviour and study of molecular interactions of chalcone in dioxane at different
concentrations and in different percentages of dioxane- water mixture at 305 K at 1 MHz
frequency have been investigated by Pathare[16]. The present work deals with the effect of
different percentage of Dioxane-water mixture on acoustical parameters of synthesized Schiff
base ligand at 30 ± 0.10C.
EXPERIMENTAL
The Dibromo Chalcone ligand were synthesized in the laboratory by literature method
[17]. All the chemicals used are of good analytical grade (AR). The solution of solute was

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

prepared by dissolving required amount of ligand in different percentage of purified Dioxane-

water solvent. The acoustical parameters are determined at fixed concentration of solute 1 x
10-3 M. The density of solvent and solutions were measured by specific gravity bottle having
10 ml capacity. The ultrasonic velocities were measured by using ultrasonic interferometer
having frequency 2MHz (Mittal Enterprises Model No. F-80).The constant temperature was
maintained by circulating water through the double wall measuring cell, made up of glass.
THEORY AND CALCULATION
The sound velocity of ligand was measured in different percentage of dioxane-water
mixture. The wavelength of ultrasonic wave is calculated using relation,
2D = λ --------------------------------------- (1)
Where, λ is wave length and D is distance in mm. The ultrasonic velocity is calculated by using
relation,
Ultrasonic velocity (U) = λ x Frequency x 103 ---------------------------------- (2)
Some acoustical parameters have been calculated using the standard relations. The
adiabatic compressibility (βs) of solvent and solution are calculated by using equations
Adiabatic compressibility solution (βs) = 100/ U2s x ds -------------------------- (3)
Adiabatic compressibility solvent (β0) = 100/ U20 x d o -------------------------- (4)
Acoustic impedance (Z) = Us x ds ------------------------------------------ (5)
Where, U0, d0, β0 and Us, ds, βs are ultrasonic velocity, density and adiabatic compressibility
of solvent and solution respectively.
Intermolecular free length (Lf) = K x √βs ------------------------------------------- (6)
Relative association (RA) = (ds /d0) x (U0 /Us)1/3 -------------------------------- (7)
Where, K is Jacobson’s constant[19] is calculated by using relation,
K= (93.875+0.375xT) x 10-8 ---------------------------------------------------- (8)
Where, T is temperature at which experiment is carried out.
Apparent molal compressibility (Øk) has been calculated by using the relation,
βs d0 – β0 ds = βs x M
(Øk) = 1000 X βs d0 – β0 ds / m x ds x d0 + βs x M/ ds ---------------------------- (9)
Apparent molal Volume (Øv) has been calculated by using the relation,
M (d0 – ds) x 1000 (Øv) = M/ds X (do-ds) x 1000/mxdsxdo ------------------------ (10)
Øk(s) = Øk0(s) + Sk(s) √m ---------------------------------------------------- (11)
Øv = Øvo + Sv √m ---------------------------------------------------------------- (12)
RESULT AND DISCUSSION
In the present investigation, different acoustical properties such as ultrasonic velocity
(Us),density (ds), adiabatic compressibility (βs), intermolecular free length (Lf), specific
acoustic impedance (Z), apparent molal volume (Øv), apparent molal compressibility(Øk(s)),

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relative association (RA) are listed in table. It is found that ultrasonic velocity decreases with
decrease in percentage of dioxane The ultrasonic velocity is related to intermolecular free
length. The variation of ultrasonic velocity in solution depends upon the increase or decrease
of intermolecular free length after mixing the solute is based on a model for sound propagation
proposed by Eyring and Kincaid[20]. The intermolecular free length (Lf) increases with
increasing the percentage of dioxane, hence decrease in ultrasonic velocity with increasing the
percentage of dioxane. This indicates that there is a weak interaction between ion and solvent
molecules which suggesting a structure non-promoting behavior of the added solute. This also
implies increase in number of free ions showing the occurrence of ionic dissociation due to
weak ion-ion interactions.
The value of specific acoustic impedance (Z) decreases with decrease in the percentage
of dioxane for given system. This is due to decrease in the strength of intermolecular attraction.
The specific acoustic impedance (Z) is directly proportional to ultrasonic velocity and inversely
proportional to adiabatic compressibility and shows similar behaviour to that of ultrasonic
velocity and opposite to that of adiabatic compressibility. It was found that the value of
adiabatic compressibility increases with decrease in percentage of dioxane indicates that there
is weak solute-solvent interaction may be due to departure of solvent molecules around ions.
The relative association is the property used to understand the interaction in solution. The
relative association is measure of extent of association of the component in the mixture. The
value of relative association decreases with decrease in percentage of Dioxane-water solvent..
This is due to the breaking up of the associated solvent molecules on addition of dioxane in it.
It was observed that apparent molal volume decreases with decrease in percentage of Dioxane-
water solvent the system. It indicates the existence of weak ion-solvent interaction.The
apparent molal volume increases due to decreasing dielectric constant of medium with increase
in percentage of Dioxane-water solvent.
The value of apparent molal compressibility decreases with decreases in percentage
of Dioxane-water solvent for the system. The positive value of apparent molal compressibility
shows the strong electrostatic attractive force in the vicinity of ions causing electrostatic
solvation of ions
Table
Acoustic Properties of ligand (L) at different percentages in Dioxane-water Mixture
% of Us ds Βs Lf Øv Øk(s) RA Z
Dioxane (cm3mole- (cm.sec-
(m.sec-1) (g.cm- (bar-1) (A0) (cm3mole- 1
bar-1) 1
.cm-3)
3 1
) )

100 1630.22 0.9039 5.04 x 10-4 13.51 x 102 1006.11 25.36 0.9176 14773.55

95 1584.30 0.9125 4.77 x 10-4 13.14 x 102 900.18 15.92 0.9194 1467.57

90 1560.45 0.9275 4.52 x 10-4 12.80 x 102 723.23 10.42 0.9285 1456.59

85 1544.15 0.9385 4.35 x 10-4 12.56 x 102 595.86 6.92 0.9354 1449.18

80 1514.65 0.9481 4.20 x 10-4 12.33 x 102 487.63 3.42 0.9410 1436.04

75 1480.85 0.9510 4.03 x 10-4 12.08 x 102 455.46 1.09 0.9479 1408.28

CONCLUSION
The present study shows the experimental data for ultrasonic velocity, density at 30
± 0.10C for synthesized Dibromo Chalcone in 70% Dioxane-water mixture. From experimental

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

data, the acoustical properties were calculated. The solute-solvent interaction and ion-ion /
solute-solute interaction existing between Dibromo Chalcone ligand and different percentage
of dioxane were also studied with the help of experimental data. Lastly it has been concluded
from the experimental data, that the solute-solvent interaction in Dibromo Chalcone and
dioxane systems are weak.

ACKNOWLEDGEMENT
The Author are thankful to Principal,vidarbha mahavidyalay,amravati for providing
necessary facilities and for kindly cooperation.
REFERENCES
[1] K. Balaramamoorthy, V. A. Chandramouli and N. KondalRao, J. Sci. Ind, Res.,32(12),1973,747.
[2] Y. Tanaka, M. Ido, Y. Umeki and S. Honda, Bull. Jp. Soc. Prec. Eng.,1975,9(4), 99.
[3] D. A. Bell, Br. Brit. Cer. Trans., 1989,88.
[4], 133. [4] O. Doutres, Y. Salissou, N. Atalla and R. Panneton, Appl. Acous., 2010,71, 506..
[5] M. Gupta and J. P. Shukla, Indian J Pure Appl Phys., 1996), 34, 772.
[6] Pankaj and C. Sharma, Ultrasonics, 1991,29, 344.
[7] S. BalajaandS. Oza, Fluid Phase Equlibria, 2002, 200, 11-18.
[8] S. S. Aswale, D. T. Tayade,S. R. Aswale and P. B. Raghuwanshi, Proceedings of 1st International
Society Bio-Technolog Conference,Gangtok, 2008, 325.
[9] S. D. Thakur,D. T. Mahajan and M. L. Narwade,J. Ind. Chem. Soc., 2007,84(5), 480..
[10]S. A. Ikhe,P. R. Rajput and M. L. Narwade,Ind. J. Chem.A, 2005,44(12), 2495.
[11] G. V. Ramana,E. Rajagopal and N. M. Murthy,J. Pure Appl. Ultrason., 2005,27,98..
[12] Raut A. R., Mhaske S. T. and Murhekar G. H. (2015), Int. J. Curr. Res. Chem., Pharma. Sci.,2(10),
79-83..
[13] Deshmukh A. O. and Raghuwanshi P. B. (2015), Int. J. Chem. Tech. Res., 8(10), 375-382. [14]
Thorat S. A. and Thakur S. D. (2015), Sci. Revs. Chem.
Commun.,5(2), 57-61..
[15] Deosarkar S. D., Jahagirdar H. G. and Talwatkar V. B. (2010), Rasayan J. Chem., 3(4), 757-760..
[16] PathareUnnati A. (2016), J. Appli. Chem., 9(2), 68-70.
[17] S. S. Lamani Kumar,OblennavarKotresh,M. A. Phaniband and J. C. Kadakol, E. Journal of
Chemistry, 2009, 6 (S1), S239S246..
[18] Vogel’s “A Text Book of Practical Organic Chemistry” (1989), V th edition, Longman Scientific
and Technical, John Willey and Sons, New York, 401.
[19] K. Sreekanth, D. Sravanakumar, J. Chem. Pharma Res., 2011,3(4), 29-41.
[20] Eyring H. and Kincaud J. F. (1938), J. Chem. Phys., 6, 620.

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8
Synthesis, Physicochemical evaluation of heteryl amino derivatives of bis[5-
cyano-1,6-dihydro-6-imino-2-isopropyl-4-(methylthio) pyrimidine] diazene
Girish Deshmukh*1 Avinash Thakare2, Chanda Gawande3
1, 2. Shankarlal Agrawal Science College, Salekasa
3. S.Chandra Mahila Mahavidyalaya, Amgaon, Maharashtra (INDIA)
Email: [email protected]

Abstract:
Study of the synthesis and pharmacological evaluation of heteryl amino derivatives of
bis[5-cyano-1,6-dihydro-6-imino-2-isopropyl-4-(methylthio)pyrimidine]diazene,we obtained
good result Antimycobacterium activity and Antityphoid activity.

Introduction:
The understanding of principles of drug design and development of the drug molecules
is important study the physicochemical properties of chemical compounds used to develop
novel pharmacologically active compounds. The biological activities, mechanism of actions,
possible biological activities of the metabolites and significance of stereochemistry for
molecules are important factors for new drug design [1]. All these principles are based on the
basic organic chemistry, physical chemistry and biochemistry. Heterocyclic compounds
contain, one or more atoms of other elements apart from carbon, common hetero atoms are
sulphur, nitrogen and oxygen [2]. The heterocyclic compounds having less common atoms
such silicon, bromine, phosphorus, tin, boron are much investigated in recent years. The
heterocycles with five or six atoms in the ring are the most important [3]. The practice of
medicinal chemistry is devoted to the research in development of new diseasetreating agent.
The process of finding a new drug is complex and involves talent of people from variety of
disciplines [4]. The important aspect of medicinal chemistry is to establish a relationship
between pharmacological activity and chemical structure [5].
Pyrimidine is a six membered cyclic compound containing 2 nitrogen and 4 carbon
atoms which is pharmacologically inactive however its substituted derivatives shows an
important place in modern medicine [6]. Pyridazine 1, oxygenated derivative-pyridazinone 2
and benzfused pyridazine or phthalazine 3 are heterocyclic compounds that contain two
adjacent nitrogen atoms (1,2-diazine) in the ring structure [7]. They show a high range of
pharmacological activities and are found in different natural compounds with different
biological activities [8]. Many heterocyclic compounds obtained from synthetic as well as
natural sources, generally in practice have one or more nitrogen in the heterocyclic ring system.
Diazines (1,2/1,3/1,4) are important heterocylic rings. Recently, much attention has been
focused on diazine derivatives for their broad-spectrum pharmacological activities [9].

Present Work:
The newly synthesised molecule possesses two methylthio groups which can be easily
substituted with heteryl amines that can be marked.
Heteryl amino derivatives ofcan be prepared by reacting it with substituted heteryl
amines in 1:2 proportion using DMF solvent and anhydrous K2CO3 catalyst to yield compounds
.

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

H H H
H HN N N NH
HN N N NH
N N K2CO3/DMF N N
+ N N
N N Reflux N
NC CN
NC CN
SCH3 N N
SCH3

N : HN HN HN O
, ,

Scheme: Synthesis of heteryl amino derivatives of bis[5-cyano-1,6-dihydro-6-imino-2-

isopropyl-4-(methylthio) pyrimidine] diazene
Results and Discussion:
In the study of optimization of reaction condition and solvent for the synthesis of
heteryl amino derivatives of bis[5-cyano-1,6-dihydro-6-imino-2-isopropyl-4-(methylthio)
pyrimidine] diazene we have obtained low yield at room temperature than the reaction carried
out at the reflux condition.
Experimental Section:
All chemicals procured from spectrochem, Alfa Aeser, SDfine, Sigma Aldrich were
used without further purification. The IR spectra were recorded with Shimadzu FTIR. The
melting points were recorded using Veego digital melting point apparatus. The NMR spectra
were recorded with Bruker 400MHz using DMSOd6 solvent.
General Procedure:
Compound (0.442 g, 1 mmol) and different heteryl amines, (1 mmol) in 1:2
proportion using 15 ml of DMF as solvent and catalyst anhydrous K2CO3as catalyst (10 mg)
reflux for 5-7 hours, the progress of reaction was monitored by TLC. The reaction mixture was
cooled to room temperature and poured into ice cold water. The separated solid product was
filtered off, washed many times with water and recrystallized from ethanol.
Spectral Data:
1) bis[5-cyano-1,6-dihydro-6-imino-2-isopropyl-4-(piperidinyl) pyrimidine]
diazene

HN
CN
N
NC HN
N N
N N N
HN N
H

 Molecular Formula : C26H36N12

 Mol. Weight (g/mol) :516.6
 -1
IR(KBr) (cm ) : 3300(=NH), 2198 (CN), 1616 (C=N)
 1H NMR (δ, ppm) : 1.24 (q, 8H), 1.36 (s, 12H), 2.82 (t, 8H), 3.86 (s, 2H),
(DMSOd6) 9.21(s, 2H)
 Mass (m/z) : 372.2, 516.4 (M+.)
2) bis[5-cyano-1,6-dihydro-6-imino-2-isopropyl-4-(p-piperazinyl) pyrimidine]

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

diazene

H
N
HN CN
N
NC HN
N N NH
N N N
HN N
H

 Molecular Formula : C24H34N14

 Mol. Weight (g/mol) :518.6
 -1
IR(KBr) (cm ) : 3309 (=NH), 2204 (CN), 1610 (C=N)
 1H NMR (δ, ppm) : 1.35 (s, 12H), 3.08 (t, 8H), 3.10 (t, 8H), 3.82 (s, 2H),
(DMSOd6) 8.00 (s, 2H)
 Mass (m/z) : 332.6, 518.9(M+.)

3) bis[5-cyano-1,6-dihydro-6-imino-2-isopropyl-4-(4-thiomorpholino)
pyrimidine] diazene

S
HN
CN
N
NC HN
N N
N N S
N
HN N
H

 Molecular Formula : C24H32N12S2

 Mol. Weight (g/mol): 552.7
 IR(KBr) (cm-1) : 3301 (=NH), 2198 (CN), 1606 (C=N)
 Mass (m/z) : 477.1, 552.6 (M+.)

Table: Synthesis of heteryl amino derivatives of bis[5-cyano-1,6-dihydro-6-

imino-2-isopropyl-4-(methylthio) pyrimidine] diazene

Reaction
Yield M.P.
Entry Substrate Product Time
(%) (0C)
(h)
O
H HN CN
N N
NC HN
N
5.0 74 194
N O
N N N
a O
HN N
H

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

HN CN
N
NC HN
N N 5.5 70 186
N N N
N
b H HN N
H

HN
CN
N
NC
N
HN
N 6.5 67 180
N N N
c N HN N
H H

H
N
H
N HN CN
N
NC HN 4.5 78 203
N N
N N N N NH
d H HN N
H

HN
NH2 N CN
NH
N NC HN
N NH 5.0 79 178
e N N N
N
HN N
H

S
H HN CN
N N
NC HN
N
6.5 83 182
N S
N N N
f S
HN N
H

Table : Physicochemical evaluation of heteryl amino derivatives bis[5-cyano-1,6-

dihydro-6-imino-2-isopropyl-4-(methylthio) pyrimidine] diazene
Solvent: DMSO
Density Viscosity Refractive
Compound
Sr. No. (ρ) ( μ*103) Index
(0.1M )
(g/cm3) poise (n)

1 3.19a 0.9520 6.3465 1.5423

2 3.19b 0.9689 6.8745 1.5279

3 3.19c 0.9588 7.9472 1.5364

4 3.19d 0.9637 6.3120 1.5457

5 3.19e 0.9697 7.1857 1.5663

6 3.19f 0.9884 6.4412 1.4666

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References:
1. Zanatta, N.; Flores, D. C.; Madruga, C. C.; Flores, A. F. C.; Bonacorso, H. ;
Tetrahedron Lett.2006, 47, 573.
2. Amir M., Javed SA, Kumar H ;Indian J. Pharm. Sciences, 2007, 69(3): 337.
3. Jain M. K., Sharnevas SC,Organic Chem. 2008, 3: 997.
4. Sasada T, Kobayashi F, Sakai N, Konakahara T; Org. Lett.2009, 11, 2161.
5. Zhichkin P, Fairfax D J, Eisenbein S A; Synthesis, 2002, 720.
6. Ahmad O. K., Hill M D, Movassaghi M; J. Org. Chem., 2009,74: 8460.
7. Barthakur M. G., Borthakur M, Devi P, Saikia C J, Saikia A, Bora U; Synlett, 2007,
223.
8. Karpov S. and Mulle T J J; Synthesis, 2003, 2815.
9. Movassaghi M., Hill M. D.; J. Am. Chem. Soc., 2006, 128: 14254.
10.GhorabM. M., Heiba MI, Hassan AA; JACS.2011,7(1),1063.
11. Kappe C. O.; Eur J Med Chem.2000, 35(12), 1043.
12. Brown D. J. New York: Pergamon press, 1984. 57.
13. Callery P., Gannett P.; Philadelphia Lippincott Williams and Wilkins, 2002. 934.
14. Polak A., Scholer HJ. ;Chemotherapy. 1975, 21, 113.
15. Cheng C., Roth B. In: Ellis GP; 8th ed. London: Butterworths, 1982. 267.
16. Ghomi A. S., Ali M.; Dig J Nanomater Biostruct 2010,5(2):303.
17. Lagoja I. M.; Chem Biodivers2005, 2(1):1.
18. Brown D. J., Evans R. F., CowdenUK; Pergamon Press Oxford; 1984.15.
19. Elderfield R. C. John Wiley & Sons New York; 1957. 6.
20. Agarwal O. P. Krishna Prakashan Media (P) Ltd.; 2006. 735.
21. Porter A. E. Pregamon Press, Elsevier Science BV Amsterdam; 1979. 14.
22. Gavilan M. D., Gomez-Vidal J. A., Serrano F. R.;Chem Lett2008; 18, 1457.
23. Ali M., Azad M., Siddiquia H. L., Nasim F. H.; J Chinese Chem Soc,
2008; 55, 394.
24. Siddiqui A. B., Trivedi A. R., Kataria V. B., Shah ;Bioorg Med Chem Lett,2014
15, 24, 1493.
25. S. Cao, X. Qian, G. Song, B. Chai, and Z. Jiang; Journal of Agricultural and Food
Chemistry,2003, 51, 152.
26. K. Abouzid, M. Abdel Hakeem, O. Khalil, and Y. Maklad; Bioorganic &
Medicinal Chemistry, 2008,16, 1, 382.
27. W. Malinka, A. Redzicka, O. Lozach, ;Il Farmaco, 2004,59, 6, 457.
28. Jain, K. S.; Chitre, T. S.; Miniyar, P. B.; Kathiravan, M. K.; Bendre, V. S.; Veer, V.S. J.
Curr. Sci. 2006, 90, 793.
29. Botta, M.; Corelli, F.; Maga, G.; Manetti, F.; Renzulli, M.; Spadari, S.;
Tetrahedron2001, 57, 8357.

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

Novel Synthesis and Characterization of Some Substituted

Chlorosubstituted Aryl Substituted 1,3-Thiazole as Antibacterial Agents

Chhaya D. Badnakhe1
1
Department of Chemistry, Dr.Manorama and Prof.H.S.Pundkar, Arts, Commerce and Science College,
Balapur, Dist. Akola.
E-mail : [email protected]

Abstract :- The synthesis, spectral analysis and biological activities of 5-(2’-hydroxy-3’,5’-

dichlorophenyl)-4-heptan-1-one-2-[4-(2-hydroxy-3,5-dichlorophenyl)-2-mercapto-
imidazolo]-1,3-thiazole (8d2) (J”) have been carried out. In this case 5-(2’-hydroxy-3’,5’-
dichlorophenyl)-4-(heptan-1-one)-2- amino-1,3-thiazole (8d) (J), 5-(2’-hydroxy-3’,5’-
dichlorophenyl)-4-heptan-1-one-2-[(2-hydroxy-3,5-dichlorophenyl) ethanonylamino]-1,3-
thiazole (8d1) (J’) & 5-(2’-hydroxy-3’,5’-dichlorophenyl)-4-heptan-1-one- 2-[4-(2-hydroxy-
3,5-dichlorophenyl)-2-mercapto-imidazolo]-1,3-thiazole (8d2) (J”) have been screened. The
compound ( J) was synthesized from 1-(2’-Hydroxy-3’,5’-dichlorophenyl)-2-bromo-1,3-
nonanedione (a4 ) by the action of thiourea, while (J”) was synthesized from (J) by reaction
with α-bromo, 2-hydroxy-3,5 dichloroacetophenone to get 5-(2’-hydroxy-3’,5’-
dichlorophenyl)-4-heptan-1-one-2-[(2-hydroxy-3,5-dichlorophenyl)ethanonylamino]-1,3-
thiazole (8d1) (J’). Further (J’) on treatment with KSCN was dissolved in acetic acid gave
(J’’). The nanoparticles of the compounds J, J’and J’’ have been prepared by using ultrasonic
technique. The newly synthesized titled compound and it’s nanoparticles were screened for
their antibacterial activity against some pathogens ; Gram+ve bacteria viz. Staphylococcus
pneumoniae, Staphylococcus aureus and Gram-ve bacteria viz. Escherichia coli and
Pseudomonas fluorescens by using agar disc diffusion method.All the newly synthesized
compounds were found to be active against test pathogens.
Keywords : Chalcone, thiazine, thiourea, α-bromo,2-hydroxy-3,5 dichloroacetophenone,
KSCN was dissolved in acetic acid, antibacterial assay.

INTRODUCTION : Heterocyclic nucleus plays an important role in medicinal chemistry

and it is a key template for the growth of var ious therapeutic agents. Thiazole is a heterocyclic
compound featuring both a nitrogen atom and sulfur atom as part of the aromatic five-
membered ring. Thiazoles and related compounds are called1,3-azoles (nitr ogen and one other
hetero atom in a f ive-membered ring). They are isomeric with the 1,2-azoles, the nitrogen and
sulphur containing compound being called isothiazoles. Thiazoles are found naturally in the
essential vitamins. Molecules that possess sulfur atoms are important in living organisms.
Chalcones and their analogues having α, β -unsaturated carbonyl system are very versatile
substrates for the evolution of various reactions and physiologically active compounds. Plant
Pathology or Phytopathology deals with the cause, etiology, resulting losses and control or
management of the plant diseases.
. It is the scientific study of diseases in plants caused by pathogens (infectious organisms) and
environmental conditions (physiological factors). Organisms that cause infectious disease
include fungi, oomycetes, bacteria, viruses,phytoplasmas, protozoa, nematod-es and parasitic
plants.
The researchers(1-6) have reported the synthesis of several thiazoles and also their potent
biological activities such as antimicrobial7, antibacterial8, antifungal9, fungicidal10 and

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insecticidal agent11.
Now a days nanotechnology is a promising field of interdisciplinary research. It opens up a
wide array of opportunities in various fields like medicine, pharmaceuticals, electronics and
agriculture. Since the physiochemical properties of nanoforms vary greatly, it becomes
important to examine the effect of nanoparticles on microorganisms to harness the benefit of
this technology in the plant protection especially against phytopathogens. Previous studies
confirmed that metal nanoparticles are effective against pathogens, insects and pests. Hence
nanoparticles can be used in the preparation of new formulations like nanomedicines for the
diseases like diagnosing & treating cancer12, enhancing outer membrane of living cells13,
inhibiting tumour growth in human being14, brain cancer 15. Nanotechnology has the potential
to revolutionize the different sectors of agriculture and food industry with modern tools for the
treatment of diseases by providing the medicines for rapid diseases like malaria16, cancer &
HIV17, breast cancer18, localized diseases19.

In the present study, the chlorosubstituted 1,3-thiazole & their imidazole derivatives (J”)
have been prepared along with their nanoparticles and screened them for their antibacterial
activities against some Gram+ve bacteria viz. Staphylococcus pneumoniae, Staphylococcus
aureus and Gram-ve bacteria viz. Escherichia coli and Pseudomonas. All the newly synthesized
compounds were found to be active against test pathogens.
EXPERIMENTAL :-
All the glasswares used in the present work were of pyrex quality. Melting points were
determined in hot paraffin bath and are uncorrected. The purity of compounds was
monitored on s ilica gel coated TLC plate. IR spectra were recorded on Perkin-Elmer
spectrophotometer in KBr pelletes, H 1 NMR spectra on spectrophotometer in CDCl3 with
TMS as internal standard. UV spectra were recorded in nujol medium. The analytical data
of the titled compounds was highly satisfactory. All the chemicals used were of analytical
grade. All the solvents used were purified by standard methods. Physical characterisation
data of all the compounds is given in Table 1.

2’-Hydroxy 3,5’-Dichloroacetophenone:
2-Hydroxy- 5-chloroacetophenone was dissolved in acetic acid (5 ml) , Sodium acetate (3g)
was added to the reaction mixture and then chlorine in acetic acid reagent (40 ml; 7.5 w/v)
was added dropwise with stirring. The temperature of the reaction mixture was maintained
be low 200 C. The mixture was allowed to stand for 30 minutes. It was poured into cold water
with stirr ing. A pale yellow solid then obtained was filtered, dried and crystallized from
ethanol to get the compound 2’-hydroxy 3’,5’-dichloroacetophenone.
Preparation of 2’-hydroxy-3’,5’-dichloro-4-hexylchalcone :
2-Hydroxy-3,5-dichloroacetophenone (0.01 mol) dissolved in ethanol (50 ml) treated
with heptanaldehyde (0.1 M) at its boiling temperature. Aqueous sodium hydroxide solution
[40%, 40 ml] was added dropwise and the the mixture was stirred mechanically at room
temperature for about 1 hour. It is then kept for 6 to 8 hours followed by decomposition with
ice cold HCl [1:1]. The yellow granules thus obtained were filtered, washed with 10%
NaHCO3 solution and finally crystallized from ethanol-acetic acid solvent mixture to get the
compound .
Preparation of 1-(2’-hydroxy-3’-5’-dichlorophenyl)-2,3-dibromononan-1-one (a1) :
2’-Hydroxy-3’,5’-dichloro-4-hexylchalcone (0.01 M) was suspended in bromine–

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

glacial acid reagent [25% w/v] [6.4 ml]. The reagent was added dropwise with constant
stirring. After complete addition of reagent the reaction mixture was kept at room
temperature for about 30 minutes. The solid product, thus separated, was filtered and washed
with a little petroleum ether to get the compound (a 1 ).
Preparation of 2-(4”-hexyl)- 6,8-dichloroflavone (a 2 ):
1-(2’-Hydroxy-3’,5’-dichlorophenyl)-2,3-dibromo-nonan-1-one (0.01 mol) was
dissolved in ethanol (25 ml). To this, aqueous solution of KOH (25 ml) was added. The
reaction mixture was refluxed for 1 hour, cooled and diluted with water. The product, thus
separated, was filtered and crystallized from ethanol to get the compound (a 2 ).
Preparation of 1-(2’-hydroxy-3’,5’-dichlorophenyl)-1,3-nonanedione (a3) :
2-(4”-Hexyl)-6,8-dichloroflavone (0.01 mol) was dissolved in ethanol (25 ml). To
this, aqueous solution of HCl (25 ml) was added. The reaction mixture was then refluxed for
one hour, cooled and diluted with water. The solid product, thus obtained, filtered and
crystallized from ethanol to get the compound (a3).
Preparation of 1-(2’-hydroxy-3’,5’-dichlorophenyl)-2-bromo-1,3-nonanedione (a 4 ) :
1-(2’-Hydroxy-3’,5’-dichlorophenyl)-1,3-nonanedione (0.01 mol) was dissolved in a
mixture of ethanol (10 ml) and dioxane (10 ml). To this, calculated amount of liquid bromine
(0.5 ml) was added. The product was not separated even after standing for one hour. It was
then diluted with water and washed with water several times and extracted with ether. The
solvent was removed under reduced pressure to get the white solid of the compound (a 4 ).
Preparation of 5-(2’-hydroxy-3’,5’-dichlorophenyl)-4-(heptan-1-one)-2- amino-1,3-
thiazole (J) :
1-(2’-Hydroxy-3’,5’-dichlorophenyl)-2-bromo-1,3-nonanedione (a 4 ) (0.01 mol) and
thiourea (0.01 mol) were dissolved in ethanol (25 ml). To this, aqueous KOH solution (0.01
mol) was added. The reaction mixture was then refluxed for three hours, cooled, diluted with
water and acidified with conc HCl. The product, thus separated, was filtered and crystallized
from ethanol to get the compound (J).
Preparation of 5-(2’-hydroxy-3’,5’-dichlorophenyl)-4-heptan-1-one-2-
[(2-hydroxy-3,5-dichlorophenyl)ethanonylamino]-1,3-thiazole (J’):
A stoichiometeric mixture of 5-(2’-hydroxy-3’,5’-dichlorophenyl)-4-(heptan-1-one)-
2- amino-1,3-thiazole (J) and α-bromo-2-hydroxy-3,5-dichloro acetophenone was dissolved in
ethanol and refluxed for one hour. It was then cooled, diluted with water and crystallized from
ethanol to get the compound (J’).
Preparation of 5-(2’-hydroxy-3’,5’-dichlorophenyl)-4-heptan-1-one- 2-[4-(2-hydroxy-
3,5-dichlorophenyl)-2-mercapto-imidazolo]-1,3-thiazole (J”):
A stoichiometeric mixture of 5-(2’-hydroxy-3’,5’-dichlorophenyl)-4-heptan-1-one-2-[(2-
hydroxy-3,5-dichlorophenyl)ethanonylamino]-1,3-thiazole(J’) and KSCN was dissolved in
acetic acid, refluxed for 4.5 hours, cooled ,diluted with water,and solid product, thus obtained
crystallized from ethanol to get the compound (J”).

The UV, IR, and NMR spectral data :-

Compound (J) :
UV : Spectrum No. 1
The UV-Vis spectrum of the compound (J) reported in dioxane showed max value 410 nm
corresponding to n transition.

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IR (KBr) :- Spectrum No. 2

3036.60 cm-1 (-OH phenolic), 2955.55cm-1 (aliphatic -C-H stretching), 3036.60cm-1 (aromatic
-C-H stretching), 3797.72 cm-1 (-NH2 stretching), 1538.48 cm-1 (-C=N stretching), 1228.56
cm-1 [(C-N=) stretching], 756.57 cm-1 (C-Cl stretching in aliphatic), 1073.66 cm-1 (C-Cl)
stretching in aromatic).

PMR :- Spectrum No. 3

∂ 5.2 (hump, 2H, -N-H2) ; ∂ 6.7 (d, 1H, -CH=C-H) ; ∂ 6.8 (d, 1H, -CH=C-H) ; ∂ 7 to
7.8 (m, 6H, Ar-H) ; offset (region not observed, observed, O-H)

Compound (J’’) :

UV : Spectrum No. 4
The UV-Vis spectrum of the compound J” reported in dioxane showed max value 399 nm
corresponding to n transition.

IR (KBr) :- Spectrum No. 5

1649 cm-1 (=C=O stretching), 3391 cm-1 (-OH phenolic), 2925 cm-1 (aliphatic -C-H stretching),
3068 cm-1 (aromatic -C-H stretching), 1435.8 cm-1 (-C=N stretching), 1305 cm-1 [(C-N) (C-
NO2) stretching], 738 cm-1 (C-Cl stretching in aliphatic), 2547 cm-1 (-S-H stretching).
1.02

PMR :- Spectrum No. 6

∂ 7.4 to 8.25 (m, 4H, Ar-H) ; ∂ 0.80 (t, 3H, -CH2-CH3, ∂ 1.064 (envelope of CH2, 10H, -(CH2)-CH3).

Preparation of Nanoparticles of the Titled Compound:

Ultrasonic Processor Sonapros PR-250MP was used to produce nanoparticles of the test
compound. The test compound was dissolved in dioxane to prepare 0.1 M solutions. This
solution was taken in a beaker and the probe of the sonapras 250 MP was dipped in solution.
These solution was exposed to sonopros MP 250 for 10 minutes separately. The test compound
was converted to nanoparticles. The solvent dioxane was evaporated by conventional heating
method. The size of nanoparticles of the test compound was confirmed by X-ray diffraction
studies using Benchtop x-ray diffraction (XRD) instrument (Miniflex).
The thin film of the nanoparticles of the test compound was prepared on glass slide. This
slide was intr oduced to the X-ray diffraction instrument to get graphical information which
was used for the calculation of the crystal size of test compounds.

Characterisation of Size of Nanoparticles of the Test Compound:

The crystal size of nanoparticles of the test compound is calculated by using Debye -
Scherrer equation.

0.94 λ
D = -------------
. Cos 

53
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

Where,
D = The average crystalline size.
0.94 = The particle shape factor which depends on the shape and size of the particle.
λ = is the wavelength.

maximum intensity peak

rad).

Spectrum No. 01

RC SAIF PU, Chandigarh

73.8
72
3797,72
70 1955,72
901,71 485,71
614,70
68 933,70 513,70
1003,69
631,68
66 1027,67
1073,66
64
1702,65
62

60
%T 3036,60 1196,60
58
2858,60
56 3208,58
1347,56 756,57
1256,56
54 1228,56
2955,55 1595,54
52 2928,54 1447,54 1316,53
696,53
50

48 1496,50

46 1538,48
45.0
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
cm-1

Chhaya D-12.sp - 8/25/2014 - L

Spectrum No. 02

54
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

J
BRUKER
AVANCE II 400 NMR

7.7079
7.4577
7.4514
7.3829
7.3767
7.3670
7.2850
7.2271
7.1594
7.1223
7.1137
7.0710
6.9896
6.9695
6.9442
6.8914
6.8160
6.7692
5.2323
3.9860
3.2603
3.1776
2.9032
2.8078
2.7908
2.7612
2.6806
2.6395
2.6017
2.5689
2.5099
2.4852
2.4698
2.4398
2.4219
2.3551
2.3200
2.3158
2.3116
2.2301
2.2023
2.1865
2.1683
2.1478
2.1129
2.0976
2.0812
2.0468
1.9891
1.9702
1.8973
1.8600
1.7199
1.5667
1.5294
1.3623
1.0646
1.0414
0.8846
0.8670
0.8496
0.6692
0.6647
0.2355
0.1953
0.1671
Spectrometer
SAIF
Panjab University
Chandigarh

Current Data Parameters

NAME Aug20-2014
EXPNO 180
PROCNO 1

F2 - Acquisition Parameters
Date_ 20140820
Time 14.16
INSTRUM spect
PROBHD 5 mm PABBO BB-
PULPROG zg30
TD 65536
SOLVENT DMSO
NS 8
DS 2
SWH 12019.230 Hz
FIDRES 0.183399 Hz
AQ 2.7263477 sec
RG 512
DW 41.600 usec
DE 6.00 usec
TE 295.7 K
D1 1.00000000 sec
TD0 1

======== CHANNEL f1 ========

NUC1 1H
P1 10.90 usec
PL1 -3.00 dB
SFO1 400.1324710 MHz

F2 - Processing parameters
SI 32768
SF 400.1300768 MHz
WDW EM
SSB 0
LB 0.30 Hz
GB 0
PC 1.00

0.33

0.44

0.10

6.00

3.08
14 13 12 11 10 9 8 7 6 5 4 3 2 1 0 ppm

[email protected]

Spectrum No. 03

Spectrum No. 04
RC SAIF PU, Chandigarh

52.1
50

45 1818,48 412,48
1738,48
1775,46 458,45

40
440,42
2605,41
2728,39 605,39
35 2547,40
3016,35 912,35
2970,35 1557,34 903,36
30 2925,34
3082,31
1022,29
%T 25 3068,27 1114,27 973,29 881,27
1088,27
867,25 556,24
20
1608,21 1134,21 642,21

15
1337,15 789,15
10
1365,11 738,11
1178,10
5
1649,7 1236,7
1434,5 1305,5
0.0
4000.0 3600 3200 2800 2400 2000 1800 1600 1400 1200 1000 800 600 400.0
cm-1

Chhaya D-18.sp - 6/17/2015 - L1

Spectrum No. 05

55
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

J1
BRUKER

12.5982
AVANCE II 400 NMR

8.2583
7.9570
7.9507
7.8094
7.8031
7.5612
7.5548
7.4793

3.3976
2.5389
2.5315
2.5272
2.5228
2.5183
2.4036
2.1127
2.0983
1.9194
1.7862
1.5823
1.3960
1.3783
1.3603
1.2680
1.2365
1.1201
1.0949
1.0774
1.0599
Spectrometer
SAIF
Panjab University
Chandigarh

Current Data Parameters

NAME May28-2015
EXPNO 130
PROCNO 1

F2 - Acquisition Parameters
Date_ 20150528
Time 13.34
INSTRUM spect
PROBHD 5 mm PABBO BB-
PULPROG zg30
TD 65536
SOLVENT DMSO
NS 8
DS 2
SWH 12019.230 Hz
FIDRES 0.183399 Hz
AQ 2.7263477 sec
RG 181
DW 41.600 usec
DE 6.00 usec
TE 297.9 K
D1 1.00000000 sec
TD0 1

======== CHANNEL f1 ========

NUC1 1H
P1 10.90 usec
PL1 -3.00 dB
SFO1 400.1324710 MHz

F2 - Processing parameters
SI 32768
SF 400.1299944 MHz
WDW EM
SSB 0
LB 0.30 Hz
GB 0
PC 1.00

1.00

1.26
1.19

3.54

7.86
4.42
16 15 14 13 12 11 10 9 8 7 6 5 4 3 2 1 0 ppm

[email protected]

Spectrum No. 06
Table 1 : Characterisation data of newly synthesized compounds :
Compounds Molecular M.P. % % of element
formula in of C H N S Cl Br
0
C yield
C8H6O2Cl2 54 80 47.90/48 2.95/3 34.15/34.58
a C15H9O4NCl2 250 70 53.10/53.25 2.40/2.66 3.98/4.18 21/21.77
a1 C15H9O4NCl2Br2 72 70 36.01/36.14 1.78/1.80 2.78/2.81 14.20/14.25 32.08/32.12
a2 C15H7O4NCl2 132 60 53.14/53.57 2.07/2.08 4.13/4.16 21.03/21.13
a3 C15H9O5NCl2 117 50 50.74/50.84 2.45/2.54 3.90/3.95 20.03/20.05
a4 C15H8O5NCl2Br 78 60 41.12/41.57 1.78/1.84 3.20/3.23 16.08/16.39 18.34/18.47
J C16H20O2N2Cl2S 96 60 51.10/51.20 5.30/5.33 7.40/7.46 7.67/7.76 17.20/17.23
J’ C24H22O4N2Cl4S 94 70 49.85/50.00 3.78/3.81 4.78/4.86 5.50/5.55 24.50/24.65
J” C25H21O3N3Cl4S2 108 70 48.60/48.62 3.38/3.40 6.75/6.80 10.35/10.37 23.00/23.01

Scheme :
Cl
OH OCOCH3 OH
CH3COONa AlCl3
OH
Cl2 - AcOH
+ (CH3CO)2O 120oC + CHO-(CH2)5-CH3
Cl Cl Cl COCH3 Cl
Cl COCH3
(2a)
OH Cl (3a)
EtOH/NaOH
Br2/AcOH
OH
(CH2)5-CH3 25% (w/v)
H H EtOH / aq KOH
Cl
(CH2)5-CH3
O Cl
(4b) Br Br
O
Cl Cl (4d)
Cl

O (CH2)5-CH3 OH EtOH/liq Br2

OH NH2-CS-NH2
H EtOH
EtOH/HCl
(CH2)5-CH3 (CH2)5-CH3
Cl Cl Cl
Br
(5d) O O O O O
(6d) Cl (7d)

Cl NH2 HO
H
OH alpha-Bromo,2-hydroxy-3,5-dichloroacetophenone
S N Cl H N
Cl
(CH2)5-CH3 OH H O KSCN in CH3COOH
Cl S N
O (CH2)5-CH3
(8d) Cl
O
(8d1)
Cl
HO
HS N
Cl

Cl N
OH
S N
(CH2)5-CH3
Cl
O
(8d2)

56
National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

Where :

1) R1 = -H, -C6H5
2) R2 = -H, -C6H5
EXPERIMENTAL DETAILS AND DISCUSSION OF RESULTS :
Antibacterial Assay :
The compounds (a – J”) were screened for their antibacterial activity against Gram +ve bacteria
viz. Staphylococcus pneumoniae, Staphylococcus aureus and Gram -ve bacteria viz.
Escherichia coli and Pseudomonas fluorescens at conc. of 1000 ppm by using Agar disc
diffusion method. Ofloxacin used as a standard and chloroform as solvent control. The zones
of inhibition formed were measured in mm and are shown in Table No.2.
Table No.2- Impact of test compounds against plant pathogens :
Sample Code (Gram positive ) (Gram Negative)
Staphylococcus Staphylococcus Escherichia Pseudomonas
pneumoniae aureus coli fluorescens
a - 15 12 12
a1 - 13 - 12
a2 17 14 12 15
a3 15 - 12 25
a4 14 12 15 12
J 14 15 15 -
J’ 12 13 12 18
J” 12 - 14 25
Reference
Antibiotic (Ofloxacin) (Ofloxacin) (Ofloxacin) (Ofloxacin)

Diameter of inhibition zone (mm)

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

RESULT AND DISCUSSION :

Most of the test compounds have shown remarkable and very encouraging antibacterial
activities. A further detailed study in the light of plant pathology is advised.
ACKNOWLEDGEMENTS :
The authors are thankful to Dr.D.H.Pundkar, Principal, Dr.Manorama & Prof.H.
S.Pundkar, Arts, Commerce & Science College, Balapur, Dr.B.B.Wankhade, Principal,
Malkapur Vidnyan Mahavidyalaya, Malkapur, and Shri Shankarlal Khandelwal College, Akola
for providing help in carrying out the antibacterial activities & for providing necessary
facilities to carry out the research work.
REFERENCES :-
(1) K. Anilkumar, P.K., K. Shrivastava Rao, Reddana P., Ind. J. of chem. sec., B, pp. 1033-
1037, June 2007.
(2) Kakade B.S., "Synthesis in heterocyclic compounds Ph.D. Thesis", Nagpur University,
1983.
(3) Seiji Miwatashi, Yasayoshi Arikava, Shigonori Ohkawa,Keiko Ugo-Chem., Pharm,
Bull. 56 (8), Vol. 56, No. 8, pp.no. 1126-1137, 2008.
(4) Airody Vasudeva Adhikari, Karabasanagouda T., Ramgopal Dhanwad and G.
Parameshwarappa, Indian J. of Chem., vol. 47 B, pg.no. 144-152, Jan. 2008.
(5) Yasuda, Nobuyuki, Karikome, Michinori, Toda, Takashi, Chem., Lett. (12), 1141-2-
1995.
(6) Bhavin Sutariya, Raziya S.K., S. Mohan and S.V. Sombassiva Rao, Indian J. of
Chem. Vol. 46 B, pp.no. 884-887, May 2007.
(7) Airody Vasudeva Adhikari, Karabasanagouda T., Ramgopal Dhanwad and G.
Parameshwarappa, Indian J. of Chem., vol. 47 B, pg.no. 144-152, Jan. 2008.
(8) Mr. Prakash Anil Castelina, Dr. Jagdish Prasad, Dr. Prasanntha Naik, Vol 4/ISSUE :
3/Mar. (2014)/ISSN-2249-555 x.
(9) Khare, P.K., H. Singh, Shrivastava, Indian J. Chem., Sec., B, 875-879, May 2007.
(10) Pathan S.R., Dighe N. and S. Shinde, H.V. Asian J. of Research chem., 2 (2), April-
June 2009.
(11) Suman Adhikari, S.B. Bari, A. Samanta, Journal of Applied Research, 8, 1, 31-40
[2014].
(12) Piotr Grodzinski, American cancer society, 3 Aug. (2012) ; DOI : 10.1002/CnCr. 27766.
(13) Yue Yaun, Xijun Wang Bin Mei, Dongxin Zhan Scientific Reports 3, Article number :
3523, doi : 10.1038/srep 035 23 ; 29 November (2013).
(14) Andrei L. Gartel, Scientifica, volume (2014) (2014), Article ID 59 658, http://dx.
doi.org./10.1155/2014/596528.
(15) Dr. Ho, Nanotech advancements, Scientific hardness nanotechnology to better diagnose
and tract cancer, 120 : 2781-2783 ; doi 10.1002/Cner.28982.
(16) Patricia Urban and Zavier Fernandez Busquets, Bentham science, Current Medicinal
chemistry, ISSN 1875-533 x. Volume 22, 38 Issues (2015).
(17) Chun-Mao Lin and Tan-Yi Lu, Bentham Science, ISSN : 2212-4020.
(18) Ming Wang, Mariana Holasia, Kasim Kabirov, Aryamitra Banerjee, Cell Cycle, vol. 11,
issue 18, (2012).
(19) Oriol Planas, Thibault Gallavardin and Santi Nonell, Institut Quimic de Sarria, Universitat
Ramon L/ull Via Augusta, 390, 08017, DOI : 10.1039/C4CC 09070 E ICommun, 2015,
51, 5586-5589.


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10

Mini Review on biological aspects of 1,3,4-Thiadiazoles

Mr. N.D. Dahake*, Dr. V.H. Masand

Abstract: Thiadiazols are a significant class of heterocyclic compounds with a wide range of
uses in biological, pharmacological, and chemical synthesis. They are also well-known for their
benefits as metal chelating agents, cyanine dyes, oxidation inhibitors, and anti-corrosion
agents. Researchers from all around the world are working on this moiety because of its wide
range of uses, which has helped to enhance the chemistry of thiadiazols. The most recent
information on advancements, novel approaches, synthetic strategies, methods used for
thiadiazole synthesis, and their various medicinal uses, as well as the structure-activity
relationship of the most effective compounds and their physical characteristics, is provided in
this review article. For medicinal and organic chemists working on developing newer
compounds with thiadiazole moiety that may be safer and more effective agents, this article
can be a valuable resource.
Key words: Semi carbazide, thiadiazole, antitumour, antioxidant, anti-inflammatory.
Antibiotic Activity: It has been found that 1,3,4-thiadiazoles have a wide range of antibacterial activity.
Through its several reactivity sites, which have been outlined above, the thiadiazole ring frequently
served as a framework to which various pharmacophore compounds with comparable antibacterial
properties were connected. Quinoline, pyrazole, triazole, piperine, imidazoline, and other well-known
pharmacophore agents were known to exhibit a range of bioactivities (Schem – 1). In order to test the
in vitro antibacterial activity of 2,8-substituted quinolin-1,3,4-thiadiazoleyl acetamide derivatives
against Staphylococcus aureus, Streptococcus pyogenes, Salmonella typhimurium, and Escherichia
coli, Bhat et al.Compounds 183 and 184 demonstrated superior inhibitory effects against S. aureus at
MIC = 16 μg/mL (controlled to amoxicillin MIC = 32 μg/mL), while compounds 185 and 186
demonstrated superior outcomes against pyogenes at MIC = 16 μg/mL (controlled to amoxicillin MIC
= 32 μg/mL).

Figure 4- Scheme -1

Antiviral Activity: Research on 1,3,4-thiadiazoles' antiviral efficacy was unsatisfactory. Despite their
best efforts, the majority of the time the researchers only saw moderate or even lower activity. Triazolo
1,2,4-[3,4-b]In vitro anti-HIV-1 (IIIB) and HIV-2 (ROD) activity in MT-4 cells was significantly lower
with -1,3,4-thiadiazoles with 3,6-substituted by substituted phenyl, heterocyclic groups such as furan,
thiophene, pyrozine, pyridine, pyrrole, coumarin, etc., adamantyl and acyclic C-nucleosides (compound

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

214) than the reference drug (Scheme 2). Moreover, 2-arylthioacetamino-1,3,4-thiadiazoles were
produced and assessed for their ability to block HIV-1 reverse transcriptase as non-nucleoside inhibitors
(NNRTIs) through an allosteric interaction with the non-nucleoside inhibitor binding site, or NNBS,
which is a location next to the NRTI binding site. The new 1,3,4-thiadiazole compounds did not perform
as well as their benzoimidazole analogues and the reference drug zidovudine, despite the fact that the
nitrogen atom and the amino group of thiadiazole can form H bonds with the RT NNBS residues and
the 5-position substituent can also contribute to the interaction with NNBS observed through docking
simulation. Comparing thiadiazole compounds to their benzoimidazole equivalents and the reference
medication zidovudine, they did not perform as well.

Figure 5 - Antiviral Analogues of 1,3,4-Thiadiazole

Antioxidant Activity: Antioxidants can stop chain reactions that could harm or kill a cell by
removing free radical intermediates from oxidation events. Usually, they are reducing agents
like polyphenols, ascorbic acid, or thiols. Similar to chemical antioxidants, a network of
interacting antioxidant enzymes, including catalase, peroxiredoxins, thioredoxin and
glutathione systems, superoxide dismutase (SODs), and catalase, protects cells from oxidative
stress. There is a lot of literature on the antioxidant properties of 1,3,4-thiadiazoles. Compared
to the reference antioxidant propyl gallate, N-(2,4-Dimethylphenyl)-5-(4-nitrophenyl)-1,3,4-
thiadiazol-2-amine exhibited more superoxide anion scavenging action. The presence of a
potent electron-withdrawing group (NO2) may be responsible for stabilising the negatively
charged adduct that superoxide forms. This outcome supported the report by Chidananda.
Notwithstanding the unsurprising results, some (benzo)-imidazothiadiazoles and
triazolothiadiazoles demonstrated potential antioxidant activities against a variety of in vitro
systems, including superoxide radical, 2,2-diphenyl-1-picrylhydrazyl DPPH radical,
microsomal NADPH-dependent inhibition of lipid peroxidation LP levels, nitric oxide
scavenging activity, and microsomal ethoxy resorufin O-deethylase (EROD). When used to
scavenge free radicals (DPPH•, ABTS•+, and •OH) in vitro, thiol and aminothiol compounds
formed from 1,3,4-thiadiazoles have been shown to have much greater antioxidant properties
than their comparable analogues derived from benzothiazoles, with thiols being superior to
aminothiols In comparison to the radioprotectors WR-2721 and WR-1065, analogue provided
83% protection against 2-deoxyribose degradation by •OH at 60 μM, scavenging DPPH• and
ABTS•+ free radicals with IC50 values of 0.053 and 0.023 mM, respectively. However, in vivo
tests revealed that benzothiazoles had a more effective radioprotective effect. It was proposed
that there is a direct connection between the aromatic ring's ability to trap radicals and the thiol
function's ability to capture them. Furthermore, studies in the literature have shown that 1,3,4-
thiadiazoles had less antioxidant activity than oxadiazole units in both nitric oxide and DPPH
techniques.61a,b The mutagenicity effect of cyclophosphamide was able to be mitigated by
melatonin, the pineal gland indole, and its thiadiazol indole derivatives. This reduction in
mutagenicity effect may have been caused by the antioxidant activity of these compounds in
male mice. Additionally, the thiadiazole ring's existence was unable to interfere with its action.

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

Melatonin activity, however, was not affected by the mere existence of a thiadiazole ring fused
to a melatonin moiety with an acetonitrile side chain.

References
1. Bhat, A. R.; Tazeem; Azam, A.; Choi, I.; Athar, F. Eur. J. Med. Chem. 2011, 46, 3158.
2. Xiaohe, Z.; Yu, Q.; Hong, Y.; Xiuqing, S.; Rugang, Z. Chem. Biol. Drug Des. 2010,
76, 330.
3. (a) Limon-Pacheco, J.; Gonsebatt, M. E. ́ Mutat. Res., Genet. Toxicol. Environ.
Mutagen. 2009, 674, 137. (b) Pastore, A.; Federici, G.; Bertini, E.; Piemonte, F. Clin.
Chim. Acta 2003, 333, 19. (c) Rahman, I.; Biswas, S. K. Encyclopedia of Respiratory
Medicine; Academic Press: Oxford, 2006
4. Pandey, A., Rajavel, R., Chandraker, S., Dash, D., Eur. J. Chem. 2012, 9(4), 2524-2531.
5. Kai, Z., Wang, P., Li-Na, X., Xiao-Yun, F., Fen, J., Sha, L., Yuming, L., Bao-Quan, C.,
Bioorg. Med. Chem. Lett. 2014, 24, 5154-5156.
6. Raj, M. M., Patel, H, V., Raj, L. M., Patel, N. K., Int. J. Pharma. Chem. Bio. Sci. 2013,
3(3), 814-819.
7. Husam, A. A., Ahlam, J. Q., Iraqi. J. Pharm. Sci. 2012, 21(1), 98-104.
8. Ahmad, T., Singh, A. K., Jaiswal, N., Singh, D., Int. Res. J. Pharm. 2012, 3, 70-82.
9. Plonka, W., Paneth, A., Paneth, P., Mol. 2020, 25(20), 4645.
10. Ming, C., Wen, G. D., Gui, S. L., Zhong, T. F., Xiu, W., Mol. 2021, 26(6), 1708.
11. Georgeta, S., Mol. 2020, 25(4), 942.
12. Wadher, S. J., Puranik, M. P., Karande, N. A., Yeole, P. G., Int. J. Pharm. Tech. Res.
2009, 1(1), 22-33.
13. Akocak, S., Lolak, N., Nocentini, A., Karakoc, G., Tufan, A., & Supuran, C. T.,
Bioorg. Med. Chem. 2017, 25(12), 3093-3097.
14. Hamadneh, L. A., Sabbah, D. A., Hikmat, S. J., Al-Samad, L. A., Hasan, M., Al-Qirim,
T. M., & Al-Dujaili, A. H., Mol. Cell. Biocheme. 2019, 458(1), 39-47.
15. Akram, E., Daham, S. N., Rashad, A. A., & Mahmood, A. E., Al-Nahrain J. Sci. 2019,
22(1), 25-32.
16. Gaber, M., El-Wakiel, N., El-Baradie, K., & Hafez, S., J. Iran. Chem. Soc. 2019, 16(1),
169-182.
17. Godhani, D. R., Mulani, V. B., Mehta, J. P., & Kukadiya, N. B., World. Scient. News.
2018, 100, 51-60.
18. Rajavel, P., Senthil, M. S., Anitha, C., E-J. Chem. 2008, 5(3), 620-626.
19. Pandey, A. K., Bangladesh. J. Pharmacol. 2019, 14(3), 127-128.
20. Ibrahim, D. H., Saleem, A. J., Awad, A. A., Ahmed, H. S., & Shneshil, M. K., Insti.
Phys. Publis. 2019, 1294(5), 052029.

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11

Simultaneous Non-aqueous Conductometric Investigation of

Pharmaceutically Potent Ibuprofen-Paracetamol Combination Drugs
Pradip P. Deohate, Aakanksha S. Dongare and Anushri R. Jalamkar
Department of Chemistry, Shri Radhakisan Laxminarayan Toshniwal College of Science, Akola-444001, India
E-mail : [email protected]
Abstract
Simultaneous non-aqueous conductometric investigation of pharmaceutically potent
ibuprofen-paracetamol combination drugs has been accomplished using the solvent
isopropanol and the titrant KOH in isopropanol. Drugs ibuprofen and paracetamol are distinctly
acidic in nature. These drug combinations are extensively used in medicines and
pharmaceuticals. In present work, ibuprofen-paracetamol combination drugs are
simultaneously investigated by non-aqueous conductometric titration method using the dip
type of glass conductivity cell. This method has been found to be precise for assay of
combination drugs and can be used even in common laboratories. The results are compared to
those obtained by Indian Pharmacopoeia (IP) method.
Keywords : Non-aqueous, conductometric investigation, ibuprofen-paracetamol
Introduction
The literature survey showed that, different pharmaceutical drugs and organic
compounds have been analyzed by the conductometric method1-3. Good amount of literature is
available on determination of pharmaceutical compounds by acid-base titration method in non-
aqueous solvents as well as other methods. Procedures for non-aqueous titrations are described
in the main pharmacopoeias4-6. Conductometric titration method using conductivity cell is also
recommended by the pharmacopoeias. Determination of pharmaceutical compounds by non-
aqueous titration method is discussed and reviewed by many authors7-9. In the literature,
different methods are given for the determination of ibuprofen 10,11. Non-aqueous
conductometric estimation of ibuprofen in perchloric acid has been reported earlier12. For the
determination of combination drugs, different methods have been suggested and these are
mostly concerns with the separation of components followed by individual component
estimation using suitable technique. Investigation of pharmaceutically potent ibuprofen-
paracetamol combination drugs by conductometric titration method using solvent isopropanol
was not yet reported in literature. As the drugs ibuprofen and paracetamol are distinctly acidic
in nature, could not be titrated directly with aqueous alkali because of their hydrolysis. The
basic titrants are superior to the alkoxide solvents. These are more susceptible to the
atmospheric moisture and carbondioxide.
Herein, the easy method to investigate the pharmaceutically potent drugs is reported. It
will help in the investigation of raw materials and products for quick check of spurious drugs
that are feared to penetrate the markets. In the reported work, conductometric titrations in non-
aqueous medium using the solvent isopropanol and titrant KOH in isopropanol were carried
out to analyze the ibuprofen and paracetamol in two component combination drugs without
any separation.

Material and Methods

All the titrations were performed by a digital conductometer (Systronics-304) with dip
type of glass conductivity cell having outer diameter of 15 mm and cell constant 1.
Conductivity cell was conditioned in the desired solvent for 24 hours before the use.
Conductometric titrations were carried out at constant temperature using thermostatic bath and
the volume of titration solution was always kept constant (20 ml). Suitable volume corrections

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were applied whenever necessary. Weightings of required drugs and chemicals were made on
Contech, CAH-123 (±0.001 g) balance. Chemicals and solvents used were AR grade. Solvents
were purified and made anhydrous by standard methods13 and titrants were protected from
atmospheric moisture and CO2. Drugs used in present work were obtained from pharmaceutical
laboratories. These are pharmaceutical in nature and included in pharmacopoeias4-6.
Ten tablets of the same batch of ibuprofen-paracetamol drugs were accurately weighed
and powdered for the present investigation. As per the requirement, quantity of powder
equivalent to about 400 mg of ibuprofen and 325 mg of paracetamol was weighed accurately,
treated with 50 ml of isopropanol and stirred vigorously for dissolving the active component
of the tablets. Binding agents and fillers remained insoluble. Common additives present in the
tablets i.e. calcium carbonate, glucose, lactose, starch, gum etc. were mostly insoluble in
isopropanol. Solutions were filtered, residues were washed with small portions of isopropanol
for three to four times and volumes of solutions were made to 100 ml with isopropanol.
Aliquots of 10 ml of these solutions were diluted to 20 ml with isopropanol and titrated with
0.1 M solution of KOH in isopropanol by conductometric method using a conductivity cell.
Conductometrically, titrant was standardized with 0.1 M benzoic acid in isopropanol. End
points were found out by plotting the graphs and later on amount of drugs present in titrated
weights of tablet powder were calculated. Amount of active components (drugs) present in one
tablet was calculated from the average weight of the tablet. Same tablets were also analyzed by
the method given in pharmacopoeias so as to compare the results.

Results and Discussion

Ten tablets of the same batch of drugs ibuprofen and paracetamol were weighed and
powdered. Powder equivalent to 400 mg of ibuprofen and 325 mg of paracetamol was exactly
weighed, extracted with isopropanol and volume was made to 100 ml. The 10 ml of this
solution was diluted to 20 ml with isopropanol and titrated conductometrically with KOH in
isopropanol. Standardization of titrant was done using standard benzoic acid in isopropanol by
conductometric titration. Weight of drugs ibuprofen and paracetamol in titrated amount of
tablets was calculated. Investigation of same tablets was done by IP method and result for two
tablets of different brands was tabulated. It was observed that, the reported method of non-
aqueous conductometric titration gives quite accurate and comparable results to those obtained
by IP method (Table-1). It is simple, precise and free from indicator error or interferences.
Acidic drugs get hydrolyzed in presence of aqueous alkali, but in non-aqueous medium it can
be avoided. As per the US Pharmacopoeia, alcoholic solution of the acidic drugs is to be titrated
with aqueous alkali and such titration must be performed very fastly for minimizing the
hydrolysis. Present method has not this limitation. The most common additives present in the
tablets i.e. calcium carbonate, sugars, gum etc. are insoluble in isopropanol and do not affect
the results. Conductometric breaks obtained using isopropanol as solvent are quite pronounced
and prominent with minimum error (Graph-1). At the end point, solvent isopropanol permitted
a large change in the concentration of solvated proton. Solvent isopropanol can be purified and
made anhydrous very easily. Its dielectric constant is smaller. Present method is simple than
other one, where the components are separated and investigated by spectrophotometric,
chromatographic or other techniques.
Table-1 : Conductometric investigation of ibuprofen-paracetamol combination drugs

Sample Label Claim (mg) Weight Found by IP method (mg) Weight Found by present method (mg)

Ibuprofen Paracetamol Ibuprofen Paracetamol Ibuprofen Paracetamol

DT-01 400 325 398.80 326.80 399.10 327.50

DT-02 400 325 401.90 317.10 402.60 319.70

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Graph-1 : Conductometric investigation of ibuprofen-paracetamol combination drugs

Conclusion
Simultaneous non-aqueous conductometric investigation of pharmaceutically potent ibuprofen-
paracetamol combination drugs is fast, simple, precise method. It can be worked out in all common
laboratories without the use of any kind of sophisticated equipments. For titration of drugs in non-
aqueous medium, solvent isopropanol was found to be good one with accurate results. KOH in
isopropanol was found to be better basic titrant to the alkoxide solvents that are susceptible to
atmospheric moisture and carbondioxide.

Acknowledgement
Authors are thankful to Principal, Shri Radhakisan Laxminarayan Toshniwal College of
Science, Akola for providing sophisticated equipments and necessary laboratory facilities.

References
1. S. Ashour and H. Aboudan, Int. J. Pharm. Chem., 4(1), 8 (2018).
2. F. M. Salama, K. A. M. Altia, R. A. M. Said, A. El-Olemy and A. M. Abdel-Raoof, J. Adv. Pharm. Res.,
2(2), 113 (2018).
3. M. Ayad, M. El-Balkiny, M. Hosny and Y. Metias, Ind. J. Adv. Chem. Sci., 4(2), 149 (2016).
4. “Pharmacopoeia of India”, Directorate of Publications, New Delhi (2007).
5. “British Pharmacopoeia”, Her Majesty’s Stationary Office, London (2004).
6. “United States Pharmacopoeia XX and National Formulatory XV”, U.S. Pharmacopeia Convention,
Rockville (1980).
7. L. Safarik and M. Kasova, Pharmazie, 36, 843 (1984).
8. M. Beslova, Z. Bezakova, M. Bachrata and J. Subter, J. Farmaceuticky Obzor., 56, 97 (1987).
9. P. N. Vyas and R. B. Kharat, J. Ind. Chem. Soc., 65, 885 (1988).
10. E. M. Costi, I. Goryacheva, M. D. Sicilia, S. Rubio and D. Perez-Bendito, J. Chromatogr., 1210(1), 1 (2008).
11. R. Hamoudova and M. Pospisilova, J. Pharm. Biomed. Anal., 41(4), 1463 (2006).
12. S. Ghora, O. Halder, A. Bagchi, P. Mukherjee, A. Raha and M. Pal, Int. J. Pharm. Res. Health, 6(3), 2643
(2018).
13. J. Kucharsky and L. Safarik, “Titrations in non-aqueous solvents”, Elsevier, New York (1965).

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12
Cp*Co(III) Catalyzed Redox-Neutral Annulation: A Powerful Tool for
Isoquinoline Synthesis from N-Cbz Hydrazones
Dewal S. Deshmukh1* and Bhalchandra M. Bhanage2
1
Department of Chemistry, J. D. Patil Sangludkar Mahavidyalaya, Daryapur- 444803 (MH)
2
Department of Chemistry, Institute of Chemical Technology, Matunga, Mumbai-400019
e-mail id: [email protected]
Abstract
A new cascade oxidative cyclization reaction of N-Cbz hydrazones with internal
alkynes has been explored for the preparation of isoquinoline derivatives using Cp*CoIII-
catalyst via C-H and N-N bond functionalization. N-Cbz hydrazones are rarely explored as
directing group for redox-neutral [4+2] cyclization reaction through the cyclometallation and
this catalyst system does not require any external oxidizing agent as well as silver or antimony
salt. The demonstrated efficient approach has been utilized for the synthesis of different
isoquinoline derivatives with good regioselectivity and yields.
Introduction
Transition metal-catalyzed C-H bond functionalization is vital for modifying organic
molecules, particularly in drug synthesis and green chemistry. Current efforts are directed at
enhancing Cp*Co(CO)I2-catalyzed C-H bond activation with first-row transition metals [1-5].
Isoquinoline derivatives, essential for diverse pharmacological applications and as catalyst
ligands, are employed in materials, dye, and paint industries using various name reactions. In
recent decades, C-H bond activation has become a pivotal pathway for achieving high yields
in isoquinoline derivative synthesis [6-12].

Scheme 1. Transition metal catalyzed annulation reaction for the synthesis of isoquinoline.

Exploring transition metal-catalyzed isoquinoline synthesis has faced challenges like

external oxidant requirements, expensive catalysts, and limited starting materials. Addressing
these issues, the Zhu group showcased the use of the affordable cobalt catalyst for C-H/N-N
bond activations. [13–17]. Challenges in existing protocols, such as the use of air and silver
salt, highlight the need for a cost-effective Co-catalyzed synthetic protocol without external
oxidants [18].
Our research expedites ortho C-H bond activation, utilizing Cp*Co(III) in a redox-neutral
cyclization with the seldom-studied N-Cbz hydrazone, providing an elegant, cost-effective, and
user-friendly method for isoquinoline synthesis without the need for silver salt and with an
external oxidant.

Results and discussion

Optimizing reaction conditions using 0.5 mmol N-Cbz hydrazone and
diphenylacetylene in 2 mL trifluoroethanol with Cp*Co(CO)I2, NaOAc, and AgSbF6 resulted

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in a satisfactory 71% yield for isoquinoline 3a at 100 °C for 16 hours; further screening
revealed varying impacts of acetate sources and solvents on product yield, with the absence of
silver hexafluoroantimonate salt surprisingly having no effect on the final yield.

Raising the reaction temperature to 110°C significantly impacted product yield, while no
change was observed at 120°C. Extending reaction time to 18 hours had no effect, but reducing
it to 14 hours resulted in a lower yield. The optimal catalyst loading for the model reaction was
found to be 10 mol% (Table 1, entries 12–17).

Scheme 2. The scope of substituted N-Cbz hydrazones and Alkynes.a

Under optimized conditions, our versatile protocol produced promising isoquinoline

products (87–78% yield) from various substituted N-Cbz hydrazones, showcasing adaptability
with electron-donating and -withdrawing groups. Notably, substituting N-Cbz hydrazone
without a substitute yielded product 3a significantly, and introducing methyl and methoxy
groups increased yields (88% and 90% for 3b & 3c). Electron-deficient groups (fluoro, chloro,
bromo, nitro) yielded products with 80%, 77%, 72%, and 63% yields (3d-3g), and varied

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positions of functional groups minimally impacted yields. The reaction with diphenyl acetylene
and meta-methyl N-Cbz hydrazone resulted in a 71% yield (3h). Annulating disubstituted N-
Cbz hydrazones yielded product 3k with an 85% yield. Isoquinoline products from various
ketones (3l, 3m, 3n, 3o) showed yields of 89%, 85%, 82%, and 81%, respectively.
Diphenyl acetylene and N-Cbz hydrazone, derived from a heterocyclic ketone, effectively
yield the desired product (3p) in satisfactory yield. Exploring various N-Cbz hydrazones, we
further investigated different alkynes for isoquinoline derivative preparation, where 3-hexyne
and unsymmetrical alkynes (but-1-yn-1-ylbenzene and prop-1-yn-1-ylbenzene) produced the
corresponding products (3q, 3r, 3s) with good yields.

Conclusion
Developed a redox-neutral Cp*CoIII-catalyzed dehydrative cyclization of N-Cbz
hydrazones with alkynes, eliminating the need for silver salt or an external oxidant, yielding
multi-substituted isoquinoline derivatives efficiently.

References
[1] Jordan, F.; Szostak, M.; Nareddy, P. ACS Catal. 2017, 7, 5721-5745. DOI:
10.1021/acscatal.7b01645.
[2] Abrams, D. J.; Provencher, P. A; Sorensen E. J. Chem. Soc. Rev. 2018, 47, 8925-8967. DOI:
10.1039/C8CS00716K.
[3] Ping, Y.; Ding, Q.; Peng, Y., ACS Catal. 2016, 6, 5989-6005. DOI: 10.1021/acscatal.6b01632.
[4] Liu, W.; Ackermann, L. ACS Catal. 2016, 6, 3743-3752. 10.1021/acscatal.6b00993.
[5] Wang, S.; Chen, S. Y.; Yu, X. Q. Chem. Commun. 2017, 53, 3165-3180. DOI:
10.1039/C6CC09651D.
[6] Bentley, K. W. Nat. Prod. Rep. 2006, 23, 444-463. DOI: 10.1039/B509523A.
[7] Tsuboyama, A.; Iwawaki, H.; Furugori, M.; Mukaide, T.; Kamatani, J.; Igawa, S.; Moriyama, T.;
Miura, S.; Takiguchi, T.; Okada, S.; Hoshino, M.; Ueno, K. J. Am. Chem. Soc. 2003, 125, 12971-
12979. DOI: 10.1021/ja034732d.
[8] Fisher, N. I.; Hamer, F. M. J. Chem. Soc. 1934, 1905-1910. DOI: 10.1039/JR9340001905.
[9] Whaley, W.; Govindachari, T. In Organic Reactions, ed. R. Adams, Wiley, New York, 1951, 6, 151.
[10] Gensler, W. In Organic Reactions, ed. R. Adams, Wiley, New York, 1951, 6, 191.
[11] Whaley, W.; Govindachari, T. In Organic Reactions, ed. R. Adams, Wiley, New York, 1951, 6, 74.
[12] He, R.; Huang, Z. T.; Zheng, Q. Y.; Wang, C. Tet. Lett. 2014, 55, 5705.
DOI:10.1016/j.tetlet.2014.08.077.
[13] Sen, M.; Kalsi, D.; Sundararaju, B. Chem. Eur. J. 2015, 21, 15529-15533. DOI:
10.1002/chem.201503643.
[14] Zhang, S.; Huang, D.; Xu, G.; Wang, S.C.R.; Peng, S.; Sun, J. Org. Biomol. Chem. 2015, 13, 7920-
7923. DOI: 10.1039/C5OB01171J.
[15] Wang, Y. F.; Toh, K. K.; Lee, J. Y.; Chiba, S. Angew. Chem. Int. Ed. 2011, 50, 5927-5931. DOI:
10.1002/anie.201101009.
[16] Zhang, S. S.; Liu, X. G.; Chen, S. Y.; Tan, D. H.; Jiang, C. Y.; Wu, J. Q.; Li, Q.; Wang, H. Adv.
Synth. Catal. 2016, 358, 1705-1710. DOI: 10.1002/adsc.201600025.
[17] Zhou, S.; Wang, M.; Wang, L.; Chen, K.; Wang, J.; Song, C.; Zhu, J. Org. Lett. 2016, 18, 5632-
5635. DOI: 10.1021/acs.orglett.6b02870.
[18] Pawar, A. B.; Agarwal, D.; Lade, D. M. J. Org. Chem. 2016, 81, 11409-11415. DOI:
10.1021/acs.joc.6b02001.

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13
Design, synthesis, spectral characterization and anti-microbial screening of
some novel 1, 2, 4- dithiazole with Coumarin moiety.
K. U. Dongarea* S. A. Waghmareb, S. B. Sarkatec, R. N. Ingoled
a,b,c-
Department of Chemistry, Ghulam Nabi Azad Arts, Commerce and Science College Barshitakli Dist.
Akola 444401 (M.S.), India
d-
Department of Chemistry, Shri Vitthal Rukhmini College, Sawana, Mahagaon, Dist. Yavatmal, (M.S.), India
*
Corresponding [email protected]

ABSTRACT
One step synthesis of 7-(3-substitutedimino-1,2,4-dithiazolo)-4-methyl-2H-chromen-
2-one (IIa-e) was carried out by oxidative cyclisation of 7-(5-substituted-2,4-dithiobiureto)-4-
methyl-2H-chromen-2-one (Ia-e) by using liquid bromine in chloroform medium as an
oxidizing agent. The synthesized compounds from these reaction conditions were characterized
on the basis of elemental analysis, chemical characteristic and IR, PMR MASS, spectral data
and antimicrobial activity against gram positive and gram-negative bacteria such as S. typhi,
E. coli, S. aureus, A. aerogenes, B. subtilis and B. megatherium. etc.
Keywords: 7-(5-substituted-2,4-dithiobiureto)-4-methyl-2H-chromen-2-one, liquid bromine,
chloroform, ammonium hydroxide ethanol, gram positive and gram-negative bacteria etc.

INTRODUCTION
Organic compounds containing thiadiazol1 as the heterocycles in their structure are
identified for their biological activities, pharmaceutical activities, industrial applications,
agricultural purposes and in medicinal sciences2-3. Basically, structure of thiadiazol is five
member nitrogen and sulphur containing heterocyclic compound. Presence of sulphur and
nitrogen in its structure enhances the biological potential of the thiadiazol. Kalogirou4 had
reported 1,2,3-dithiazole for the antimicrobial activities, Laitinen et al.5 reported the 1,2,3-
dithiazole for the antibacterial activities, Matysiak6 had reported thiadiazol and some different
substituents attached thiadiazol directly or indirectly shows effect directly in their biological,
medicinal property.
Similarly, Sayed7 has prepared the thiadiazol based coumarin the best antimicrobial
activities. The biological as well as industrial applications of coumarin8 get enhanced due the
presence of the thiadiazol nucleus in their structure. This 1,2,4-dithiazole synthesis technique
is quicker, less complicated, less expensive and requires less time.
MATERIAL AND METHOD
Material
All the chemical used were of loba chemie (AR grade).
Method:
In the present experiment for the synthesis of different substituted 7-(3-
substitutedimino-1,2,4-dithiazolo)-4-methyl-2H-chromen-2-one by using liquid bromine in
chloroform medium. Reaction mixture was kept at room conditions for 4 hours.
EXPERIMENTAL
All the chemicals used for the synthesis must be purified. After refluxing the purity of
the compounds were checked by TLC (aluminium TLC) with thin layer thickness of 200 um.
The melting points of all synthesize compounds will be recorded using hot paraffin bath. The
carbon and hydrogen analysis were carried out on Carlo-Ebra-1106 analyser Nitrogen
estimation were carried out with colmon-N-analyzer-29. IR spectra were recorded with Bruker

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spectrometer in the range 4000-400 cm-1. PMR spectra were recorded on VARIAN 400 MHz
spectrometer with TMS as internal standard using CDCL3 and DMSO Solvent.
GENARAL PROCEDURE
In a clean China dish paste of 7-(5-substituted-2,4-dithiobiureto)-4-methyl-2H-
chromen-2-one (Ia-e) was prepared in chloroform and 5% liquid bromine in chloroform (2 ml)
was added with constant stirring at room conditions. During the addition of bromine solution
to firstly the colour of bromine disappear, addition of bromine was continued till colour of
bromine appears and persists to the paste. Reaction mixture was kept for 4 hours at room
conditions. On basification of the reaction mixture with dilute ammonium hydroxide solution,
pale yellow solid of (IIa-e) obtained.
Similar procedure was adopted for the synthesis of all the derivatives in the series.
The tentative reaction for the formation of product is depicted below,

Reaction
1 NH NH NH O O
R

S S

CH3
7-(5-substituted-2,4-dithiobiureto)-4-methyl-2H-chromen-2-one
(Ia-e)

Br2 CHCl3

S S
N
1 N NH O O
R

CH3

7-{[3-(substituted)-1,2,4-dithiazol-5-yl]amino}-4-methyl-2H-chromen-2-one
(IIa-e)

Similarly, 4-methyl-7-{[3-(phenylimino)-1,2,4-dithiazol-5-yl]amino}-2H-chromen-2-
one (IIb), 7-{[3-(tert-butylimino)-1,2,4-dithiazol-5-yl]amino}-4-methyl-2H-chromen-2-one
(IIc), 4-methyl-7-({3-[(4-methylphenyl)imino]-1,2,4-dithiazol-5-yl}amino)-2H-chromen-2-
one (IId) and 7-({3-[(4-chlorophenyl)imino]-1,2,4-dithiazol-5-yl}amino)-4-methyl-2H-
chromen-2-one (IIe) were synthesized from the oxidative cyclisation 7-(5-phenyl-2,4-
dithiobiureto)-4-methyl-2H-chromen-2-one (Ib), 7-(5-tert-butyl-2,4-dithiobiureto)-4-methyl-
2H-chromen-2-one (Ic), 7-[5-(4-methylphenyl)-2,4-dithiobiureto]-4-methyl-2H-chromen-2-
one (Id) and 7-[5-(4-chlorophenyl)-2,4-dithiobiureto]-4-methyl-2H-chromen-2-one (Ie) with
liquid bromine in chloroform medium respectively by the above mentioned procedure.
RESULT AND DISCUSSION
Spectral characterization results for all the synthesized compounds are given below
Spectral Characterization
1) 7-{[3-(Ethylimino)-1,2,4-dithiazol-5-yl]amino}-4-methyl-2H-chromen-2-one (IIa)
Yellow solid, C14H13N3O2S2, Yield-87%, M.P.-142 0C. %Composition found (calculated): C-
52.59(52.64), H-4.07(4.10), N-13.14(13.15), O-10.01(10.01) and S-20.03(20.07). IR spectrum
(cm-1): 3130.50 (Ar C-H stretching), 1140.12(C=S stretching), 3340.43 (NH stretching) and 1740.41
(C=O strech lactone). PMR spectrum: The spectrum of a compound was carried out in CDCl3. This
spectrum distinctly displayed the signals due to singlet 3 protons of -CH3 gives singlet at δ 2.4 ppm

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due to attachment with aromatic ring, 3 protons of -CH= of aromatic ring gives doublet at δ 7.7, 7.74
and 7.3 ppm, 1 proton of -CH= of conjugated lactone gives singlet at δ 5.84 ppm, 2 protons of -CH2
gives quartet at δ 3.5 ppm, 3 protons of -CH3 gives triplet at δ 1.38 ppm, 1 proton -NH attached
between thionyl group and phenyl ring also desheilded due to thioamide position and gives signal at
δ 3.9 ppm. Mol. Wt.: 319.40.
2) 4-Methyl-7-{[3-(phenylimino)-1,2,4-dithiazol-5-yl]amino}-2H-chromen-2-one (IIb)
Yellow solid, C18H13N3O2S2, Yield-85%, M.P.-134 0C. %Composition found (calculated): C-
58.78(58.83), H-3.53(3.56), N-11.43(11.43), O-8.70(8.70) and S-17.41(17.45). IR spectrum (cm-
1
): 3050.12 (Ar C-H stretching), 1090.06 (C=S stretching), 3400.12 (NH stretching), 1753.94 (C=O
strech lactone) and 1580.29 (NH bending). PMR spectrum: The spectrum of a compound was
carried out in CDCl3. This spectrum distinctly displayed the signals due to singlet 3 protons of -CH3
gives singlet at δ 2.4 ppm due to attachment with aromatic ring, 4 protons of -CH= of aromatic ring
gives doublet at δ 7.7, 7.3, 7.4 and 7.6 ppm, 1 proton of -CH= of conjugated lactone gives singlet at
δ 5.85 ppm, 1 proton -NH attached between thionyl group and phenyl ring also desheilded due to
thioamide position and gives signal at δ 3.8 ppm. Mol. Wt.: 367.44.
3) Synthesis of 7-{[-3-(tert-butylimino)-1,2,4-dithiazol-5-yl]amino}-4-methyl-2H-chromen-2-
one(IIc)
Yellow amorphous solid, C16H17N3S2O2, yield 85%, M.P. 110 0C, % Composition found
(calculated), C-55.33(55.33), H-4.94(4.93), N-12.09(12.09), O-9.20(9.20), S-18.45(18.45). IR
spectrum (cm-1): 3450.12 (N-H stretching), 3034.72 (C-H Ar Stretching), 1560.56 (N-H Bending),
1120.12 (C=S stretching) and 1740.40 C=O stretching(lactone). PMR spectrum: The spectrum of
a compound was carried out in CDCl3. This spectrum distinctly displayed the signals due to singlet
of 3H of –(CH3) at δ 2.42 ppm, singlet of 9H of –(CH3)3 at δ 1.35 ppm, singlet of 1H of -CH= of
conjugated lactone gives singlet at δ 5.8 ppm, 3 protons of -CH= of aromatic benzene ring gives
doublet at δ 7.7, 7.4 and 7.7 ppm and singlet of -NH at δ 3.4 ppm. Mol. Wt.: 347.45.
4) 4-Methyl-7-({3-[(4-methylphenyl)imino]-1,2,4-dithiazol-5-yl}amino)-2H-chromen-2-one (IId)
Yellow solid, C19H15N3O2S2, Yield-88%, M.P.-118 0C. %Composition found (calculated): C-
59.76(59.82), H-3.93(3.96), N-11.01(11.01), O-8.38(8.38) and S-19.77(16.81). IR spectrum (cm-
1
): 3052.15 (Ar C-H stretching), 1050.20 (C=S stretching), 3470.12 (NH stretching), 1735.12 (C=O
stretch lactone), 1570.12 (NH bending) and 2930.52 C-H (CH3 stretching). PMR spectrum: The
spectrum of a compound was carried out in CDCl3. This spectrum distinctly displayed the signals
due to singlet 3 protons of -CH3 gives singlet at δ 2.4 and 2.02 ppm due to attachment with aromatic
ring, 5 protons of -CH= of aromatic ring gives doublet at δ 7.2, 7.7, 7.03, 7.3 and 7.6 ppm, 1 proton
of -CH= of conjugated lactone gives singlet at δ 5.8 ppm, 1 proton -NH attached between thionyl
group and phenyl ring also desheilded due to thioamide position and gives signal at δ 3.5 ppm. Mol.
Wt.: 381.47.
5) 7-({3-[(4-Chlorophenyl)imino]-1,2,4-dithiazol-5-yl}amino)-4-methyl-2H-chromen-2-one (IIe)
Yellow solid, C18H12N3O2S2Cl, Yield-84%, M.P.-130 0C. %Composition found (calculated): C-
53.74(553.79), H-2.98(3.009), N-10.45 (10.45), O-7.96(7.95) and S-15.92(15.95). IR spectrum
(cm-1): 3080.45 (Ar C-H stretching), 1160.50 (C=S stretching), 3470.15 (NH stretching), 1745.12
(C=O strech lactone), 1570.01 (NH bending) and 2950.63 C-H (CH3 stretching). PMR spectrum:
The spectrum of a compound was carried out in CDCl3. This spectrum distinctly displayed the signals
due to singlet 3 protons of -CH3 gives singlet at δ 2.3 ppm due to attachment with aromatic ring, 4
protons of -CH= of aromatic ring gives doublet at δ 7.29, 7.78, 7.54, 7.35 and 7.60 ppm, 1 proton of
-CH= of conjugated lactone gives singlet at δ 5.86 ppm, 1 proton -NH attached between thionyl
group and phenyl ring also desheilded due to thioamide position and gives signal at δ 3.5 ppm. Mol.
Wt.: 401.88.
CONCLUSION
In the present research work is cheaper and less time consuming method for synthesis of
organic compound (IIa-e). In all the synthesized compounds give the maximum yield of
product (II a-e). A variety of coumarin based 1,2,4-dithiazole derivative can be synthesised for
their antimicrobial activities adopting the method.
PHARMACOLOGICAL STUDIES
Antimicrobial Activity

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

All the synthesized compounds (II-a) to (II-e) were screened for antibacterial activity
against S. typhi, E. coli, S. aureus, A. aerogenes, B. subtilis and B. megatherium. by disc
diffusion method was performed using Mueller Hinton agar as well as nutrient agar medium.
Each and every compound was tested at conc. 50 μg/ml in ethanol. The zone of inhibition of
all the synthesized compounds were measured after 24-hour incubation at 37 0C. Standard
drug used for comparison the activity was Ciprofloxacin.
Table 1: Synthesized compound IIa-e activity against Gram +ve and Gram -ve bacteria.

Table No. 1.1– Antimicrobial sensitivity test against bacteria (after 24 hours at 37 0C
temp.)
(zone of inhibition in mm)
SR Gram positive Gram negative
No Compd. S. B. B. A.
S. typhi E. Coli
. aureus subtilis megatherium aerogenes
11 KUD-IIa 12 00 12 12 00 12
12 KUD-IIb 14 12 14 14 12 16
13 KUD-IIc 12 14 00 14 00 16
14 KUD-IId 14 14 11 14 14 11
15 KUD-IIe 18 14 11 16 12 12
Standard
27 18 20 22 24 16 12
Ciprofloxacin

Anti-microbial activities of 7-(3-substitutedimino-1,2,4-dithiazolo)-4-

methyl-2H-chromen-2-one
30

25

20

15

10

0
KUD-IIa KUD-IIb KUD-IIc KUD-IId KUD-IIe Standard
Ciprofloxacin

S. aureus B. subtilis B. megatherium S. typhi E. Coli A. aerogenes

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REFERENCES

1. Ibrahim, S. A., Salem, M. M., Abd Elsalam, H. A., & Noser, A. A. (2022). Design, synthesis, in-
silico and biological evaluation of novel 2-Amino-1, 3, 4-thiadiazole based hydrides as B-cell
lymphoma-2 inhibitors with potential anticancer effects. Journal of Molecular Structure, 1268,
133673, 1-14.
2. Kollu, U., Avula, V. K. R., Vallela, S., Pasupuleti, V. R., Zyryanov, G. V., Neelam, Y. S., &
Chamarthi, N. R. (2021). Synthesis, antioxidant activity and bioinformatics studies of L-3-
hydroxytyrosine templated N-alkyl/aryl substituted urea/thioureas. Bioorganic chemistry, 111,
104837, 1-13.
3. Tayade, D., T., Kale, P., R., Waghmare, S., A. & Nagrale, A., P. (2016). Studies in the
antimicrobial activities of some newly synthesized [1,4]substitutedbenzodiazepines by Disc
diffusion method and MIC Method, Haya: The Saudi Journal of Life Sciences, 1(2), 72-75.
4. Kalogirou, A. S., Oh, H. J., & Asquith, C. R. (2023). The Synthesis and Biological Applications
of the 1, 2, 3-Dithiazole Scaffold. Molecules, 28(7), 3193, 1-33.
5. Laitinen, T., Baranovsky, I. V., Konstantinova, L. S., Poso, A., Rakitin, O. A., & Asquith, C. R.
(2020). Antimicrobial and Antifungal Activity of Rare Substituted 1, 2, 3-Thiaselenazoles and
Corresponding Matched Pair 1, 2, 3-Dithiazoles. Antibiotics, 9(7), 369, 1-15.
6. Matysiak, J. (2015). Biological and pharmacological activities of 1, 3, 4-thiadiazole based
compounds. Mini reviews in medicinal chemistry, 15(9), 762-775.
7. Sayed, M. T., Elsharabasy, S. A., & Abdel-Aziem, A. (2023). Synthesis and antimicrobial activity
of new series of thiazoles, pyridines and pyrazoles based on coumarin moiety. Scientific
Reports, 13(1), 9912, 1-9.
8. Chandrakanth, M., Thomas, N. M., Arya, C. G., Fabitha, K., & Banothu, J. (2023). Coumarin–1,
2, 4-Triazole Hybrids: Recent Advances in Synthesis and Medicinal Applications. Journal of
Molecular Structure, 137197, 1-36.
9. Al-Warhi, T., Sabt, A., Elkaeed, E. B., & Eldehna, W. M. (2020). Recent advancements of
coumarin-based anticancer agents: An up-to-date review. Bioorganic Chemistry, 103, 104163, 1-
15.
10. Fordyce, E. A., Morrison, A. J., Sharp, R. D., & Paton, R. M. (2010). Microwave-induced
generation and reactions of nitrile sulfides: an improved method for the synthesis of isothiazoles
and 1, 2, 4-thiadiazoles. Tetrahedron, 66(35), 7192-7197.
11. Nandgude, T. D., Bhise, K. S., & Gupta, V. B. (2008). Characterization of hydrochloride and
tannate salts of diphenhydramine. Indian journal of pharmaceutical sciences, 70(4), 482.
12. Maffuid, K. A., Koyioni, M., Torrice, C. D., Murphy, W. A., Mewada, H. K., Koutentis, P. A., ...
& Asquith, C. R. (2021). Design and evaluation of 1, 2, 3-dithiazoles and fused 1, 2, 4-dithiazines
as anti-cancer agents. Bioorganic & Medicinal Chemistry Letters, 43, 128078, 1-5.
13. Gupta, A., Mishra, P., Kashaw, S. K., & Jatav, V. (2008). Synthesis, anticonvulsant, antimicrobial
and analgesic activity of novel 1, 2, 4-dithiazoles. Indian journal of pharmaceutical
sciences, 70(4), 535-538.

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14

Effect Of metal-ligand complexon germination of some vegetable plants

Prof. M.M.Rathore and C.N.Deshmukh
Department of Chemistry,VidyaBharti Mahavidyalaya, Amravati

Plant physiology will probably also assume an increasingly important role in

agricultural research problems. As world population increases, mankind faces enormously
complex problems. One of the primary tasks of the future will be to increase food, forage, fiber
and wood production substantially throughout the world. Today the application of various
chemical salts to soils is a basic feature of agricultural practice.
In the present work, Chalcone(an α,β-dihydroxy Ketone)treatment on Vegetable plant’
is selected for study as they have both nutritional as well as medicinal value. Since organic
drugs have intense biological activity and since no work is reported on the biological
applications of binary complexes of Fe (III) with ligand(chalcone) and comparing with pure
ligand, metal and control solution doubly distilled water to study the effect of complex, metal,
ligand and control solution on germination survival, seedling height, etc. on fenugreek and
Spinish plants in order to make suggestion whether complex, metal and ligands can be used
as plant growth regulators.
The following aspects were studied in laboratory.
1) Estimation of Root / Shoot Ratio in soil & soilless media.
2) Estimation of chlorophyll contents in soil & soilless media.

EXPERIMENTAL:-
The information about the role of metal complexes in biological systems, their
concentration and presence in different equilibria is of immense importance. Greshon et. al.1,
2 reported that the activity of metal chelates is considerably increased as compared to that of
the free metal and ligand alone on their complexation. The Shel et. al.3 and Shashindharam et.
al.4 observed the antifungal and antibacterial activities of complexes shows that they are more
active as compared to free ligand and metal involved.
Rare earth ions are used as probe in bio-chemistry of calcium. Zielinski et. al.5 showed
that, Lanthanide ion could substitute the calcium ion to produce active enzyme system. Some
bivalent metal ions have been reported to be useful in agriculture as plant growth regulators.
Such a vast uses of lanthanide necessitate concentrating on the study of lanthanides and ligands
for studying the germination pattern.
The complexes of transition metal with bis-alkyl thiourea are prepared and their
herbicidal and plant growth regulating activity are tested with wheat and cucumbers by Darnall
et. al.6.
Sayed amir et.al.7 studied effect of some heavy metals on seed germination of canola,
wheat, safflower evaluate phytoremedial potential.
K. Abraham et. al.8 also studied effect of heavy metals on seed germination of archis
hypogeae. L
Shivakumar C. K. et. al.9 also observed the presence of beneficial fungus and effect of
copper and zinc metal absorption on growth and metal uptake of legumineous plants as
although these metals are required in traces but are important for growth.
A. Ramteke et. al.10 studied the effect of chlorosubstituted pyrrazole and their complexes

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

on spinach at different pH.

Recently J.G.Wu et. al11 18, 23 studied effect of complex on plants.

SYNTHESIS OF CHALCONE BY KNOWN METHOD

THE CHALCONE WAS PREPARED BY KNOWN LITERATURE METHOD AND WAS CONFIRMED BY
MELTING POINT AND ALSO THE STRUCTURE WAS CONFIRMED BY IR SPECTROSCOPY AS
SHOWN IN SPECTRA .
Step I: Preparation of p-cresyl acetate from p-cresol

Step II:Preparation of 2-hydroxy-5–methylacetophenone from p-cresyl acetate

Step III:Preparation of chalcone from 2-hydroxy-5-methyl acetophenone

MATERIAL AND EXPERIMENTAL METHODS

• Metal ions:-
The solutions of metal ion in the form of FeCl3 and MgCl2 of 0.1 M concentration
were prepared using distilled water and the seeds of fenugreek and Spinach were soaked
in both metal solvents.
• Ligand:-
The organic compound was prepared were dissolved in proper solvent and
above seeds are soaked for 2-3 hours.

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

Metal ion + Ligand:- The mixture of FeCl3 and organic compound (chalcone) and
MgCl2 and organic compound (chalcone) were dissolved in the distilled water and seeds
are soaked.
Media:-For the germination of the above vegetables seeds, two types of the medias
are used.
1) Soil media (media A)
2) Soilless media (media B)

EXPERIMENTS PERFORMED :-
In general practice, various chemicals are used in agricultural as an important ingredient
of various pesticides, insecticides, fertilizers, etc. to improve the crop yield. Amongst several
economical important plants Fenugreek and Spinach are selected as a plants system. These
plants are in ideal systems to study the germination and growth pattern. Further, their
economical importance is reflected by its wide use for food. The important uses these plants
in daily life are persuasive to study its response against metal ion, ligand and its complex
regarding to physiological processes; particularly germination is a vital process for the growth
of plants. Therefore, these plants are selected as a plant system.

1]Healthy seeds were taken and thoroughly washed using doubly distilled water. Seeds from
these healthy seeds of equal size were chosen, immersed in tested solution. These seeds soaked
were taken out of each solution the seeds are sown in germination trays of all medias.

2]Effect of ligand, metal Fe (III), complex and metal Mg (II), complex on chlorophyll in the
leaves of vegetables plants were studied.
After sufficient growth, green fresh leaves were collected, as they contain
chlorophyll pigments and chlorophyll content was determined spectrophotometrically given by
12
Jahagirdar D. V.

ESTIMATION OF CHLOROPHYLL IN LEAF PIGMENT:-

Procedure:- Collect the fresh leaves weigh around 1 gm of leaves and cut them into small
pieces. Add 5 ml of water and transfer the mixture to a blender. Homogenize the mixture by
blending it intermittently for 3-4 minutes.Take 0.5 ml of homogenous mixture and add 4.5 ml
of 80% acetone to it. Thoroughly shake the content, centrifuge it. Collect the supernatant
liquid and measure its optical density at 645 and 663 nm.

• Parameters:-Plants growth is decided on the basis of parameters such as percentage

of germination survival, seedling height, shoot length, root length (root length / shoot
length) and thickness of young leaf having high values compared to control system.
The germination was noted after7days ,14 days and 22days for all Plants. After
noteing the survival of the plants, they were taken out of the medias. The seedling
height (root length / shoot length) was measured. The average values of these
parameters are presented in Table 1

RESULT AND DISCUSSION

Some attempts have been made by Bera et. al.13 to study the effect of tannery effluent
on seed germination, seedling growth and chloroplast pigment content in mungbean.
Adhikari et. al.14 have observed the effect of raw savage water on mustard. Recently Farzin

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

M. Parabia et. al.15 in their present investigation, effect of ligand, complex and metal ion on
percentage seed germination, root length, shoot length (root / shoot ratio) has been studied.
ROOT LENGTH, SHOOT LENGTH AND ROOT / SHOOT RATIO

Germination starts when the seed shows emergence phase of growth, which begins,
with penetration of embryo from the seed coat and end with development of root and shoot
system. The elongation of shoot axis follows emergence of radical.
The rate and extent of elongation is subjected to a variety of controls, including
nutrition, hormones and environmental factors. Though the root and shoot evelopment start
within a fraction of time but the further developments may vary according to the nutrients
required for the development of root and shoot independently. Therefore, root and shoot length
differs from each other.

Table 1:-Effect of Different Treatment on Vegetable Plant in Respect of Parameters in Soil

and
Soilless Media.

Sr.N Parameters Fenugreek Spinach

o.
Soil Soil
Soilless Soilless
1 % Germination 736% 86% 88% 90.4%
2 Seedling 11.96 12.2 9.77 10.6
Height(cm) 6
3 Shoot Length(cm) 6.44 6.46 6.04 6.29
4 Root length(cm) 5.52 5.76 3.74 4.28
5 Shoot/Root Ratio 1.17 1.15 1.68 1.50

Table 1 clearly indicates that percent germination in soilless medium is higher

followed soil media. Similarly the root length and shoot length which is called as seedling
height shows a significant development of root / shoot length i.e. height of seedling highest in
chalcone + Mg as compaired to over all the treatments and subsequently followed by chalcone
+ Fe, metal ion Mg, chalcone and control (d/w) respectively in all cases. When we compare
the performance of all the treatments for different parameters in soil and soilless media. The
germination & growth parameters are studied in soil and soilless media. Soilless media shows
better performance as against soil media in all the treatments.
Mg is a major constituent for the formation of chlorophyll molecule which helps in the
process of photosynthesis for the production of food materials in the plant i.e. sugar
synthesis.With combination of chalcone + Mg plays a pinnacle role in keeping all the system
working properly. This may be the reason for the better performance.
Fe plays major role in energy transfer within the plants and also brings about
chlorophyll development and formation. It is also a constituent of certain enzyme and protein.
With the combination of chalcone plays a major role in keeping all the plant system working
properly may be the reason for a good performance.
The germination of seed and development of the seedling is better in soilless media
than the soil media. Because there is less resistance for the root development and shoot

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

development in soilless media than the soil media may be the reason for better, overall
development of the plant.

• CHLOROPHYLL CONTENT
Photosynthesis is the process in which the light energy will be converted into chemical
energy. There are some basic requirements for the process of photosynthesis as CO2, H2O and
light energy besides of course, the structural framework of green plant in the form of
chloroplast, which is a unique cell having most important role in all the physiological reactions,
starting from the absorption of light energy. Basically, among the smallest group of
coordinating pigment molecules necessary to effect a photochemical
act, the most important pigments involved in photosynthesis are chlorophyll and carotenoid.
Table 2-3 clearly shows that absorption of plant leaves is higher at 663 nm in all the
treatments. These tables also clearly indicates that the amount of chlorophyll is more in
chalcone + Mg. Followed by chalcone + Fe, metal ion Mg, chalcone and control (d/w) in both
soil and soilless media.
Obviously the chlorophyll content is highest in chalcone + Mg in both soil and
soilless medias. Because of Mg is major constituent for the formation of chlorophyll.

Table 1:-Effect of different treatment on chlorophyll content in respect to soil media

Sr.No. Nameof Treatment Treatment Treatment Treatment Treatment

vegetable with with with with Metal with
plants Ligand Ligand+Mg Ligand+Fe ion Mg distilled
water
1 Fenugreek 8.22x10-3 10.162x10- 8.363x10-3 11x10-3 5.64x10-3
3

3 Spinach 4.71x10-3 7.65x10-3 5.61x10-3 8.09x10-3 4.50x10-3

Table 2:-Effect of different treatment on chlorophyll content in respect to soilless media

Sr.No. Nameof Treatment Treatment Treatment Treatment Treatment

vegetable with with with with Metal with
plants Ligand Ligand+Mg Ligand+Fe ion Mg distilled
water
1 Fenugreek 11.9x10-3 14.8x10-3 12.5x10-3 19.3x10-3 10.3x10-3

3 Spinach 7.41x10-3 8.45x10-3 8.31x10-3 9.93x10-3 5.41x10-3

When we compare the performance of all treatments for different parameters, in soil
and soilless medias. It is clearly observed that soil less media shows better result in respect of
germination percentage, development of seedling during the experimental period.

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Result of effect of metal ion Mg, Chalcone, Chalcone+ Mg and control (d/W) and
chalcone+ fe, on germination, seedling development clearly revels that, Chalcone +Mg shows
significant better performance overall the treatments. All the parameters are considered, while
finding out the results. In general order in all the parameters performance wise Chalcone+Mg
stood first followed by metal ion Mg, Chalcone Chalcone+fe and control (D/W).
The germination and growth parameter are studied in soil and soilless media. The
soilless media shows better performance as against the soil media in all the treatments.
Mg is major constituents for the formation of chlorophyll molecule which helps in the
process of photosynthesis for the production of food materials in the plants ,i.e sugar synthesis
with the combination of Mg chalcone plays a pinnacle role in keeping all the plant system
working properly may be the reason for a better performance.
Mg is a secondary importance element essential for the plant growth. Which is also a
constituents of many enzyme the detail information about Mg is already mention in the above
para. So, keeping in view of all the characteristics of Mg plays a pivotal role for a good
performance against the other treatments.
Germination of seed and development of seedling is better in the soilless media than
the soil media, because there is less resistance for the root development and shoot development
on the soilless media than the soil media, may be the reason for better overall development of
the plant.
The analysis perform for finding out the total chlorophyll in green leaves of the plant. The
results of analysis clearly indicates that metal ion Mg is having highest chlorophyll content in
both soil and soilless media than the remaining treatment like Chalcone+Mg, Chalcone+fe ,
Chalcone and control (D/W).

REFERENCES

1] Greshon, H; Parmegiani, R., and Nicerson, W. J. : Appl.

Microbiol., 10, 556 (1962).
2] Greshon, H., Parmegiani, R. : Appl. Microbiol., 11, 62 (1963).
3] Shel, A. M., Shariel, E. A., Gharib, A. and Ammar, Y. A. :
J Ind.Chem. Soc., 60, 1067 (1968).
4] Shashindharam, P. and Ramchandra, L. K. : J. Ind. Chem. Soc.,
62, 920 (1985).
5] Zielinski, S. Lomosik, L. and Wojclechowska, A. : Mh. chem.,
245, 6484 (1970).
6] Darnell, D. W. and Brinhawn, E. R. : J. Bio. Chem., 245, 6484
(1970).
7] Sayed Amir et. al. : J.of Agri. Science, vol (4) No.9, (2012)
8] K. Abrahm et. al.: peparment of environment of science S. U.
University Tirupati.
9] Shivakumar C. K.: International multidisciplinary research
journal vol(2), 06-12,(2012).
10] A. A. Ramteke et. al.: J. of chem.. and phrm. Research vol(4)
1889-1894, (2012).
11] Jahagirdar, D.V. : Experiments in Chemistry, 1st Edition,
Himalaya Publishing House (1994).
12] Bera, A. K. and Bokaria, K. :Envir. Ecol., 17(4), 958 (1999).
13] Adhikari, S., Mitra, A. and Gupta, S. K. : J. Instr. Publ. Hlth.
Engrs., India, 2, 5 (1998).
14] J.G.Wu, R.N.Den, Z.N.Chen: Trans. Met.Chem.18, 23 ,2020

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15
Preparation of Adsorbent from various natural materials for
removal of heavy metal ions from waste water: Critical review.
Sandhya B. Gaikwad
([email protected]),
MVP SAMAJ'S K.T.H.M. COLLEGE, Nashik
(K.R.T. Arts, B.H. Commerce and A.M. Science College.)

Abstract
Anthropogenic and industrial activities release heavy metal ions into the water. They may be
poisonous or carcinogenic in nature and has serious risks to both aquatic environments and
people. Heavy metal removal from wastewater is a severe issue as a result. The adsorption
method is advantageous for removing heavy metals from wastewater due to its accessibility,
affordability and eco-friendliness. High removal capacity of commercial adsorbents and bio
adsorbents are both utilized to remove heavy metals from wastewater. This review article
intends to gather erratically collect data on the many adsorbents used for heavy metal removal
and to provide details on the commercially available and natural bio adsorbents utilized in
particular for the removal of copper, lead, Nickel and cadmium.
KEYWORDS: Biosorption, Bio adsorbent, Heavy Metals ions.
1. Introduction
Environmental contamination is one of the major issues of this century as a result of industrial
development and the manufacturing of diverse industrial products around the world. [1] Heavy
metal mercury can accumulate more quickly in cropland and bodies of water as a result of
mining, smelting and sewage irrigation. Through sewage irrigation, mercury can go to crops,
which could be harmful to human health. [2]A significant amount of chromium is found in the
waste water from chromium mining, smelting, manufacturing, planting, metal processing,
tanning, paints and pigments, printing and dyeing. Recently there has been a lot of interest in
studying how to use agricultural waste products as adsorbents to remove heavy metals from
industrial effluents. A component of biotechnology is the use of agricultural by-products in the
process known as bio-sorption to remove heavy metal ions from the environment. This is
acknowledged as a new method for treating heavy metal-polluted streams. The metals of
immediate concern, according to the World Health Organization are Cr (III), Cr (VI), Zn, Cd,
Cu, Ni, Hg, Pb, Al and Mn. [4]Environmental pollution brought on by human activity has grown
dramatically over the past few decades and has reached alarming levels in terms of its negative
effects on living things. [5] The higher concentration of heavy metals causes adverse changes in
color, test, odor of water and it also stains clothes and utensils. The Overload of heavy metals
may cause severe health problems such as liver cancer, diabetes, cirrhosis of liver, disease
related to heart and central nervous systems, infertility, etc. [10]

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

2.Material and Methods: -

Sr Name of Heavy Initial pH Contact Adsorbent Maximum Langmuir Referen

no adsorbent metal conc. time in amount adsorption isotherm fit ce
. used ion minuts/hr. capacity
removal ppm or
mgl-

1 Soybean oil Cd, Zn, 100-300 1-5 1-60 0.02-2gm T.Maha

residue Pb mudion
Cd=40min o(2022)
Zn=20min

Pb=10min.

2 1.Biochar(B Hg2+ 0.5-20 11.722 Y.Bai

) and J.
,9.152 Hong
2.Modified (2021)
biochar 2hr
(MB)

3.Bentonite
(BE)

4.Biochar –
bentonite
composite

3 Wheat Cr(VI) 200 3 125.6 Langmuir Y.Chen

straw, (2020)
Cr(III) 5 68.9
Corn straw
87.4

62.3

4 1.Carbonize Cr 1.5 2-10 20min- 0.2-0.6 S.M.

d corn husk 40min Batagar
Ni Agitation awa
2.Uncarboni rate(100-
sed corn Mn 300 rpm (2019)
husk

3.carbonised
Millet husk

4.
uncarbonize
d Millet
husk

5.carbonized
husk

6.Uncarbonz
ed husk

5 Wheat Cu 5.55 5 98% N.N.

Bandela
Bajra 5.6 (2016)
Sugarcane 5.8

corn 5.6

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

6 Carbonized Pb(II) 40min 50mg 98% FTIR S.M

milet husk, 100rpm .Batagar
Cd(II)Ni 100rpm 94% SEM awa
(II) (2017)
94%
Acid
treated
corbonised
millet husk
97%

92%

94%

7. Modified Zn2+ 4 6 70 400 167mgg-1 Langmuir B.U.

Nanokaoline Mahesw
(NK), 143mgg-1 Freundlich, ari
(2019)
Modified Temkin
Nanobenton
Dubnin
ite (NB)
Raushkevic
h

SEM

3.Discussion
The synthetic and industrial waste water samples were treated by using different natural
materials to remove different types of heavy metal ions. These natural materials are adsorbents
are simple to produce and their raw materials are widely accessible. Hence, it is a green
technology that significantly improves the wastewater treatment process. In general, the most
practicable approaches are chemical, adsorption, and membrane techniques. There should be
more study done on the introduction of low-cost materials and techniques for heavy metal
removal from wastewater. Future study should consider the best methods for achieving
effective metal recovery with minimal negative effects on the environment and low costs.
4.Conclusion
One such method that not only addresses the removal of heavy metals from wastewater but is
also eco-friendly and has a small environmental impact is adsorption. Adsorbents like activated
carbon are commonly employed, although their use is limited by their expensive price. As a
result, it's important to consider sustainable solutions that also address the problem's wider
implications. Agricultural wastes, industrial wastes, and biochar are inexpensive, easily
accessible, renewable, and have a strong affinity for heavy metals, the use of these low-cost
bio sorbents is used. As a result, the availability of agro-wastes as a cheap method for removing
harmful heavy metals from waste water (5). The adsorbents are simple to produce and their raw
materials are widely accessible. Hence, it is a green technology that significantly improves the
wastewater treatment process.
5. Acknowledgement
Further work on creating more affordable adsorbents may aid in the remediation of heavy
metals.

6. References
T.Mahamudiono,D.Bokov,G.Widjaja,I.S.Konstantinov,K.Setiyawan,W.K.Abdelbasset, H.S.Majdi,
M.M.Kadhim, H.A.Kareem, K.Bansal- Removal of heavy metals using food industry waste as a cheap adsorbent
DOI:https://doi.org/10.1590/fst.111721, (2022).
Y. Bai and J. Hong- Preparation of a Novel Millet Straw Biochar –Bentonite Composite and its Adsorption
Property of Hg2+ in aqueous solution. Materials,14,1117.https://doi.org/10.3390/ma14051117,(2021).

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

Y.Chen,Q.Chen,H.Zhao, J.Dang,R.Jin, W.Zhao,Y.Li.-Wheat Straws and Corn Straws as adsorbents for the
removal of Cr(VI) and Cr (III) from aqueous solution :kinetics, isotherm, and mechanism. ACS Omega 5, 6003-
6009, (2020).
S.M.Batagarawa,A.K.Ajibola.-Comparative evolution for the adsorption of toxic heavy metals on to millet corn
and rice husks as adsorbents.Journal of analytical and pharmaceutical research.8,3 (2019).
N.N.Bandela, M.G.Babrekar,O.K.Jogdand,G.Kaushik.-Removal of copper from aqueous solution using local
agricultural wastes as low cost adsorbent. J.Mater.Environ.Sci.7 (6)1972-1978,(2016).
S.M.Batagarawa,L.Y.Dayo -Millet husk as efficient adsorbent for removal of Lead, Cadmium and Nickel ions
from aqueous Solution. Dutse Journal of Pure and Applied Science(DUJOPAS).3,1. (2017).
B.U.Maheswari,V.M.Siyakumar,M.Thirumarimurugan.- Synthesis, characterization of modified nano adsorbent
and its application in removal of Zn2+ ions from battery effluent.Environmental Chemistry and Ecotoxicology,1,2-
11, (2019).
A.Farhad, M.S.Hossain,T.S.Moin,S.Ahmed,A.M.Chowdhury- Utilization of waste chicken feather for the
preparation of eco-friendly and sustainable composite.https://doi.org/10.1016/j.clet.2021.100190,(2021).
H.G.Roh,S.G.Kim,J.Jung- Adsorption of heavymetal ions(Pb2+,Cu2+) on Perm-lotion –to treated human hair.
Korean J.Chem Eng.31(2),310-314, (2014).
V.Kumar,P.K.Bharati,M.Talwar,A.K.Tyagi,P.Kumar- Studies on high iron content in water resources of
Moradabad district (UP), India. Water Science 31, 44-51, (2017).
W.C.Moon,P.Alaniandy- A review on interesting properties of chicken feather as low –cost adsorbent.
International journal of integrated engineering Vol.11 No.2,136-146, (2019).

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16

Evaluation of Antioxidant Activity by DPPH Method of Thiocarbamides

P.L. Harale1, M.E. Shelke2, D.T. Tayade2

1. Research Centre and Department of Chemistry, Padmashri Vikhe Patil College, Pravaranagar.
2. Department of Chemistry, Govt. Vidarbha Institute of Science and Humanity, Amaravati.

ABSTRACT
The DPPH free radical scavenging method is recognized and reliable for the evaluation of the
antioxidant activity of chemical compounds. The antioxidant capability of thiocarbamate
compounds was characterized by its color change with DPPH (2,2-Diphenyl-2-Picrylhydrazyl)
and further analyzed its percentage scavenging activity with strong absorption at 517 nm.
Utmost thiocarbamate compounds had incomparable percentage scavenging activity and
displayed lower IC50 values (effective concentration for scavenging 50% of the initial DPPH
value (for µM) resolved excellent antioxidant activity as compared to ascorbic acid.

Keywords: Thiocarbamides, Antioxidant activity, DPPH method, IC50 value

Graphical Abstract:

1. Introduction:
Thiocarbamide compounds have wide spectrum applications in various fields such as
pharmaceutical, agriculture, medicinal, and synthetic chemistry 1-5. An antioxidant prevents the
oxidation of molecules inside a cell. Free radicals produced during the biological oxidation
chain reaction can lead to damage to a cell. Antioxidants act as reducing agents by terminating
a biological chain reaction and eliminating free radical mediates, also preventing oxidative
damage and protecting cells from severe body complaints 6-9. Researchers are constantly
working on synthesizing compounds with highly potent antioxidant activity 10-15. The DPPH
test is one of the very stable and reliable free radical scavenging methods to measure the
antioxidant activity of organic compounds 16-21. DPPH is reduced by accepting hydrogen from
hydrogen donor compounds and free radical scavenging compounds reduction shows a colour
change from violet to yellow. A study of antioxidant activity synthesized compounds was done
by calculating its % scavenging activity values and half-maximal inhibitory concentration

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values or effective concentration for scavenging 50% of the initial DPPH value (IC50 value) 22-
24.

In the present study, the antioxidant activity of novel substituted thiocarbamide compounds
was investigated by using the DPPH free radical scavenging method and comparing its
activity with ascorbic acid.

2. Experimental Section:
2.1. General
All synthesized substituted thiocarbamide compounds were purified by recrystallization with
ethanol. DPPH (2,2-Diphenyl-2-Picrylhydrazyl) and ascorbic acid of AR grade were used.
Micropipettes are used for the preparation of the microliter solution. Absorption at 517nm was
measured using a Systronic (2202 model) make Double-beam UV Spectrophotometer.

2.2. Antioxidant activity / DPPH free radical scavenging activity

Novel thiocarbamide compounds displayed antioxidant activity and were determined by using
free radical scavenging against 2,2-Diphenyl-2-Picrylhydrazyl (DPPH). 0.2 µM DPPH
solution was prepared in 95 % ethanol and used as the negative control. Then, different
substituted novel thiocarbamide compounds 200 µg/ mL were mixed with DPPH solution
which was then incubated in the dark for 30 min. at room temperature. Vitamin C is used as a
standard for comparison because of its high inhibition ability with the presence of OH group
stabilized free radical. The absorbance of the reaction mixtures was then measured at 517 nm.
By using a spectrophotometer. Plotting the % scavenging against concentration gave the
standard curve and the percentage scavenging was calculated from the following equation:
% scavenging activity = [(Abs. Blank−Abs. Sample) / Abs. Blank] ×100.
Thiocarbamides have significant antioxidant activity show low IC50 value (half-maximal
inhibitory concentration) by the DPPH method.

Table 1: Antioxidant activity expressed as IC50 values of TD5 thiocarbamides

Concentration % IC50
(µg/ml) of Scavenging value
TD5
Activity

50 45.03 1.44

100 56.12 6.57

150 65.48 11.70

200 74.44 16.84

250 85.58 21.97

300 93.73 27.11

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Table 2: Antioxidant activity of different thiocarbamides TD1-TD6

Thiocarbamide Ph-R Abs. %

Scavenging
Derivatives
200 (µM) Activity at
solution 200 (µM)

TD1 -H 0.254 72.94

TD2 -Me 0.297 68.37

TD3 -Allyl 0.295 68.58

TD4 -Ph 0.289 70.18

TD5 -PClPh 0.235 74.44

TD6 -mClPh 0.265 71.71

Vit.C 0.220 76.57

3. Result and Discussion:

Novel thiocarbamide compounds were investigated for their free radical scavenging ability in
ethanol by using DPPH assay as DPPH radical changes from Purple to yellow color when
quenched by antioxidants. DPPH radical scavenging was monitored by a Spectrophotometer at
wavelength 517 nm. Most of the derivatives showed notable antioxidant properties as
compared to Vitamin C. The Order of the activity depends on the free radical stability form in
the derivatives.
In the present research work, all the thiocarbamide compounds showed excellent
antioxidant activity with minimum IC50 values and at the lower concentration due to the
presence of the N-H group in its structure.

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Acknowledgment:
We are thankful to the Research Center of the Department of Chemistry, Govt. Vitharbha
Institute of Science and Humanity, Amaravati and Research Center of the Department of
Chemistry, Loknete Dr.Balasaheb Vikhe Patil (Padmabhushan Awardee), Pravara Rural
Education Society’s, Padmashri Vikhe Patil College, Pravaranagar.

Declaration:
Conflict of interest: The authors declare that they have no conflict of interest.
Ethical approval: This has not been published elsewhere and is not currently under
consideration for publication elsewhere

5. References:
1. Shah, Habib Ur Rehman, et al., Journal of Molecular Structure 1272, 134162, 2023.
2. Umapriya, Kollu, et al., AIP Conference Proceedings, 2390.1, 2022.
3. Al-Mulla, Abbas, Der Pharma Chemica 9.13,141-147, 2017.
4. Yanagimoto, Kenichi, et al., Journal of agricultural and food chemistry 50.19, 5480-5484,
2002.
5. Nishiyama, Tomihiro, et al., Polymer degradation and stability, 79.2, 225-230, 2003.
6. Fujisawa, Seiichiro, et al. Toxicology 177.1, 39-54,2002.
7. Beal, M. Flint., Annals of neurology 38.3, 357-366, 1995
8. Cheng, Zhihong, Jeffrey Moore, and Liangli Yu., Journal of agricultural and food
chemistry 54.20, 7429-7436, 2006.
9. Manojkumar, Parameswaran, Thengungal Kochupappy Ravi, Acta Pharmaceutica 59.2,159-
168, 2009.
10. Tsolaki, E., et al., Current Topics in Medicinal Chemistry 14.22, 2462-2477, 2014.
11. Santosh, Rangappa, et al., Chemistry Select 3.23, 6338-6343, 2018.
12. Mi, Yingqi, et al., International Journal of Biological Macromolecules 181, 572-581, 2021.
13. Radwan, Tasneem Mokhtar, et al., Journal of Heterocyclic Chemistry 57.3,1111-1122,
2020.
14. Kaddouri, Yassine, et al., Current Drug Delivery 18.3, 334-349, 2021.
15. Mahdi, Saba H., Dhuha Khudhair Rashid Al-Musawi, and Lekaa K. Abdul Karem., Journal
of Population Therapeutics and Clinical Pharmacology 30.14, 327-335, 2023.
16. Tsolaki, E., et al., Current Topics in Medicinal Chemistry 14.22, 2462-2477.17, 2014.
17. Jarallah, Shahad A., Olfat A. Nief, and Abdul Jabar Kh Atia., Journal of Pharmaceutical
Sciences and Research 11.3, 1010-1015, 2019.
18. Kizilkaya, Hakan, et al., Journal of the Chinese Chemical Society 67.9, 1696-1701,2020.
19. Shanty, Angamaly Antony, and Puzhavoorparambil Velayudhan Mohanan.,
Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy 192, 181-187,
2018.
20. Payet, Bertrand, Alain Shum Cheong Sing, and Jacqueline Smadja., Journal of
agricultural and food chemistry 53.26 (2005): 10074-10079.
21. Pontiki, Eleni, et al., Bioorganic & medicinal chemistry letters 16.8, 2234-2237, 2006.
22. Ludwig, Iziar A., et al., Food research international 61, 67-74, 2014.
23. Ouaket, Amine, et al., Mediterranean Journal of Chemistry 8.2, 103-107, 2019.
24. Lehuédé, Jacques, et al., European journal of medicinal chemistry 34.11, 991-996, 1999.

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17

Investigations of Proximate Compositions of Xanthium Strumarium L Roots

from Tisgaon (PIN Code 431002) of Taluka and District Aurangabad,
India

Jawerea Nayab Mohammad Jamil1*, Rahim Ullah Sharafat Ullah1

1: Department of Chemistry, Government Vidarbha Institute of Science and Humanities
Amravati 444 604
*: Corresponding Author
Email ID: [email protected]

Abstract
Xanthium Strumarium L showed anti-oxidant, anti-inflammatory, anti-cancer activities
as well as various medicinal significances hence it created sufficient interest to carry out
proximate investigations of roots of Xanthium Strumarium L of Tisgaon (PIN Code 431002)
of Taluka and District Sambhaji Nagar (Aurangabad), India which is hither to unknown from
this region. Recently proximate investigations of roots of Xanthium Strumarium L of Tisgaon
were successfully carried out and the present work deals with the quantitative analysis and
identification of ash content moisture content as well water solubility, acid and base solubilities
of Xanthium Strumarium L. which is important and essential part of herbal drugs in
pharmacokinetics and pharmacodynamics of that particular drug. These factors directly hamper
pharmacokinetics and pharmacodynamics of particular drugs and ultimately drug activity and
drug effect of any synthetic, semi-synthetic and herbal drugs, hence it become essential to carry
out proximate analysis of roots of Xanthium Strumarium L as they possess medicinal
applications and significances. During this study it is investigated that, the roots of Xanthium
Strumarium L. contain moisture and ash contents as well as acid-insoluble ash value, is
determination of solubilities in cold water, hot water, and 1 percent NaOH, HCl, CH3COOH
are also investigated.

Keywords: Xanthium Strumarium L, roots, proximate investigations, quantitative analysis.

Introduction
In the tropical regions of India, Xanthium Strumarium L., a member of the Composite
family, is frequently seen growing along roadsides, in fields, and in hedges. The word
"xanthium" refers to the seedpods, which change from green to yellow as they ripen (later they
become deep yellow to brown), and is derived from the ancient Greek words "xanthos," which
means "yellow," and "Strumarium," which means "cushion-like swelling." It is frequently
referred to as "chota-gokhru" because of the fruit's resemblance to a cow's toe. It is used to treat
the common condition hemicrania and is known as adhasisi in various parts of India. There are
25 species in the genus Xanthium, all of which are native to America. The medicinal plants
Xanthium Spinosum Linn. and X. Strumarium Linn. are used in Europe and North America.
Sambhaji Nagar previously known as Aurangabad is one of the oldest and historical
city in Maharashtra state of India encompass numbers of medicinal plants. Warmer climate is
good for X. Strumarium L. It is an annual herb and grows up to 1 m tall and has a short, sturdy,
hairy stem. The roots of X. Strumarium L contain tap root and secondary roots which are
irregularly lobed. Xanthium Strumarium L is traditional herbal medicine which is used to treat
bacterial, antifungal, urticaria and rheumatism infections as well as for the treatment of
arthritis, rhinitis, nasal sinusitis, headache problems by vaidu and X. Strumarium L. showed
reportable gastric ulcer, malaria, cancer, leprosy and leucoderma activies1-8. As a wider

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program of this laboratory on natural products; various proximate investigations of different

plants of various regions have been investigated in sufficient details9-13, considering all these
facts it was thought interesting to investigate proximate constituents present in roots of
Xanthium Strumarium L. of Tisgaon (PIN Code 431002) of Taluka and District Aurangabad,
India.
Materials and Methodology
All chemicals used during the research work were of A.R. grade. Freshly prepared
solutions were used throughout the research work. The solvents were purified by known
literature methods14.
Sample Preparation: The plants were collected from farms of Tisgaon of Taluka and
District Aurangabad, India in May 2023. The plants were cut along with the stem and shade
dried. Dried roots were taken in mortar pestle and crushed to prepare fine powder. This fine
powder is used for proximate investigations by known literature metods15.
Proximate Analysis
The determination of physicochemical parameters such as moisture content, total ash
value, acid-insoluble ash value, and solubility of the sample was carried out by the known
literature methods15. Solubility of the sample was checked in cold water, hot water and 1
percent NaOH, HCl, CH3COOH solution. Percentage of moisture and ash contents and acid
insoluble ash are determined by using following formula,
Loss of weight of sample
% of moisture = x 100
Weight of sample taken

while,
Percentage of solubility is determined by using following formula,
% of Solubility=(loss of weight of sample)/(weight of sample taken) ×100
The results obtained are given in Table-1
Table-1
Sr. No Proximate Parameters Loss of weight Amount of %
sample taken (in
of sample
grams)
1 Moisture content 0.220 1 22.00
2 Total ash content 0.165 1 16.50
3 Acid insoluble ash value 0.42 1 42.00
4 Coldwater solubility 0.67 1 67.00
5 Hot water solubility 0.48 1 48.00
6 NaOH solubility 0.46 1 46.00
7 HCl solubility 0.54 1 54.00
8 CH3COOH solubility 0.137 1 13.70

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Result and Discussion

Moisture content in any part of plant provides information for an activity of water-
soluble enzymes and coenzymes needed for the metabolic activities of that plant and it is
observed from Table No.-1 that, total moisture content in roots of was found to be 22.00%
which is good for metabolic activities in the plant growth and development of the plant. It was
found that the total ash content obtained from dry roots is 16.50% and acid insoluble ash value
is 42.00% which are good and these proximate parameters of plant organs are useful for the
determination of the mineral contents. Coldwater solubility and hot water solubility were found
to be 67.00% and 48.00% respectively; these proximate parameters will gave information
regarding water soluble neutral, acidic, basic and hydrocarbons present in the samples in herbal
chemistry. HCl solubility and CH3COOH solubility were found to be 54.00% and 13.70%
respectively, these proximate parameters gave information regarding basic organic
components present in the sample and
NaOH solubility was found to be 46.00% which gave information regarding acidic organic
components present in the sample.
Conclusion
The Results obtained during proximate analysis were good and it can be conclude that,
in roots of Xanthium Strumarium L. of Tisgaon (PIN Code 431002) of Taluka and District
Aurangabad, India contain good proximate parameters and the physicochemicals well as
physiological and anatomical activities of Xanthium Strumarium L. herbs at Tisgaon (PIN
Code 431002) of Taluka and District Aurangabad, India are in natural form and can be used
for medicinal purpose.

References:
1. Masvingwe . C., Mavenyengwa . M. Toxicological evaluation of the plant Xanthium strumarium
in Pigs in Zimbabwe. J Venom AnimToxins 1998, 4, 2.
2. Chopra. R.N, Nayar .S.L, Chopra I.C. Glossary of Indian Medicinal Plants. New Delhi: Council
of Scientific and Industrial Research; 1986, 259.
3. Moerman. D. Native American Ethnobotany. Oregon: Timber Press; 1998, 9.
4. Foster. S., Duke .J.A. A field Guide to Medicinal Plants. Eastern and Central N. America:
Houghton Mifflin Co., 1990, 12
5. Singh. G., Kachroo. P. Forest Flora of Srinagar, A good flora of the western Himalayas but poorly
illustrated, Some information on plant uses, 1976.
6. Kirtikar .K.R, Basu .B.D. Indian Medicinal Plants. Basu LM press,Allahabad, India.
1981:Edition 2nd, Vol.2, 1355.
7. Chopra .R.N, Nayar .S.L, Chopra .I.C. Glossary of Indian Medicinal Plants. New Delhi: Council
of Scientific and Industrial Research; 1958, 438.
8. Shivpuri . D .N, Dua . K.L. Investigations in pollen allergy in Delhi area IV clinical investigation,
Indian J Med Res, 1963, 51, 68.
9. Tayade. D .T, Shaikh R. S, Patil S. U. J. of Indian Chem Soc., 83, 1-3 (2006).
10. Shaikh. R .S, Ph. D. Thesis, Amravati University, Amravati. (2006).
11. Tayade .D .T, Shaikh.R. S, Asian J. of Chemistry”, 15, (3, 4), 1851-52 (2003).
12. Tayade .D. T, Shaikh. R.S , Indian J. applied and pure biology, 18 (2),115-157 (2003).
13. Naskari. P. N., M. Phil Dissertation, Alagappa University, Karaikudi, (2007)
14. M. Hiruy M., Bisrat. D., Asres.K., Natural Product Resources, 10, 1052-1056 (2021).
15. Sree.L.T., Vijayalakshmi K. Proximate Composition, Nutritional Evaluation and Mineral
Analysis in the Roots of an Indigenous Medicinal Plant, Alternanthera Sessilis. Int J Heal Sci Res
[Internet]. 2018;8(7):55. Available from: www.ijhsr.org

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18

Synthesis, Structural Study of Substituted

Heterocyclic Compounds

Kavita. M. Hedaa
a
Department of Chemistry,Shri R. L. T. College of Science,Akola – 444001(M.S.) India
E-mail: [email protected]

Abstract: Dithiazolidine constitutes a major role in the synthesis of various heterocyclic

moieties. They act as active precursors in synthetic heterocyclic chemistry. Synthesis of a series
of novel five member ring containing nitrogen and sulphur are well known1.A series of novel
3-thio-4-substituted-5-tolyl-[1, 2, 4]-dithiazolidine [hydrochloride] have been synthesized by
the interaction of several Ammonium aryl dithiocarbamate with N-p-tolyl-S-chloro
isothiocarbamoyl chloride in refluxing chloroform medium. The newly synthesized
compounds have been characterized by analytical and IR, 1H NMR and Mass spectral studies.

A small heterocyclic ring containing nitrogen and sulphur have been under
investigation for a long time because of their important properties. Synthesis, structural
properties and antimicrobial activities of various [1, 2, 4]-dithiazolidine have been reported
earlier2. The literature survey revealed that the [1,2,4]- dithiazolidine have been found to
possess potent anti-tumors, anti-tuberculosis3, anti-diabetic and anti-cancer4 and anti
inflammatory5 properties.
Thiocarbamides and their heterocyclic derivatives have gained recently much interest
as inhibitors of Human Immunodeficiency Virus (HIV)6 and Therapeutic agents7. Some of the
heterocyclic derivatives of thiocarbamides are found to possess diverse pharmacological
activities like antifungal and anti-tubercular agents. In view of utility of thiocarbamides, N-
aryl-S-chloro isothiocarbamoyl chloride have been used in synthesis of substituted [1, 2, 4]
dithiazolidine by interacting with Ammonium aryl dithiocabamates. The drug containing 1, 2,
4-dithiazolidines show a diverse range of physiological activities, antimicrobial8-9, anti-
inflammatory10-12, anti-ulcer13-14, and anti-cancer15. Here is reported the synthesis of several 3-
thio-4- substituted -5-tolyl-[1, 2, 4]-dithiazolidine [hydrochloride] (3a-g) have been
synthesized by the interaction of several Ammonium aryl dithiocarbamate (1a-g) with N-p-
tolyl-S-chloro isothiocarbamoyl chloride (2). The required Ammonium aryl dithiocarbamate
(1a-g) were obtained by the interaction of interaction of different amines with carbon
disulphide and Ammonia.
Results and discussion
Several 3-thio-4- substituted -5-tolyl-[1, 2, 4]-dithiazolidine [hydrochloride]
(3a-g) have been synthesized by the interaction of several Ammonium aryl dithiocarbamate
(1a-g) with N-p-tolyl-S-chloro isothiocarbamoyl chloride (2). in CHCl3. After condensation,
the solvent was distilled off to obtain a sticky residue. This residue was triturated several times
with petroleum ether (60-800C) to afford a pale yellow solid (3a-g). The product was found to
be non-desulphurrizable when boiled with alkaline lead acetate solution. The IR spectra of
products shows bands due to Ar-H, C-H, C=N, C-C, C-N, C=S, C-S, S-S stretching and
1
HNMR spectra of products distinctly displayed signals due to aromatic protons and Acetyl
protons. The Mass spectrum of product was also observed. The identities of these new 3-thio-

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4-aryl-5-tolyl-[1, 2, 4]-dithiazolidine [hydrochloride] have been established on the basis of

usual chemical transformations and also IR, 1H NMR and Mass spectral studies16-18.

Experimental
General Methods
All chemicals were research grade. Melting points determined are uncorrected.
IR spectra were recorded in KBr on a FT-IR Perkin-Elmer RXI(4000-450cm-1)
spectrophotometer. 1H NMR measurements were performed on a Bruker DRX-300 (300 MHz
FT NMR) NMR spectrometer in CDCl3 solution with TMS as internal reference. The Mass
spectra were recorded on a THERMO Finnigan LCQ Advantage max ion trap Mass
spectrometer. Thin layer chromatography (TLC) was performed on silica Gel G and spots were
visualized by iodine vapour. The compounds describe in this paper were first time synthesized
by the multistep reaction protocol.
Synthesis of Ammonium aryl dithiocarbamate19 (1a-g)
The compound Ammonium aryl dithiocarbamate was prepared by drop wise addition of Amine
[9ml] in ice cold mixture of ammonium [15ml, density 0.88] and carbon disulphide [7.5ml]
followed by the vigorous shaking. The reaction mixture was allowed to stand for 30min heavy
precipitate of Ammonium aryl dithiocarbamate separates out. Filter it and dry it.
Synthesis of N-p-tolyl-S-chloro isothiocarbamoyl chloride (2)
The N-p-tolyl-S-chloro isothioocarbamoyl chloride was prepared by the interaction of
p-tolyl isothiocyanate and by passing calculated quantity of Cl2 gas. Details of typical
experiment are as follows:
Through the chloroformic solution of p-tolyl isothiocyanate (0.002 M, 0.298 g in 15
mL) pure and dry chlorine gas (0.71g) was passed maintaining the temperature below 10oC.
Chlorine gas was generated by the addition of 35% HCl to KMnO4. After the complete addition
of chlorine gas, the resultant yellow solution was filtered to remove suspended impurities and
the clear solution was mixed with petroleum ether (60-80oC). The solvent was then removed
by distillation under vacuum. The resultant oil was again diluted with petroleum ether and
distilled under vacuum. N-p-tolyl-S-chloro isothiocarbamoyl chloride was obtained as pale
yellow oil (1.00 g).

3a:- Synthesis of 3-thio-4-substituted-5-tolyl-[1, 2, 4]-dithiazolidine [hydrochloride]

A mixture of Ammonium phenyl dithiocarbamate (1a-g) and N-tolyl-S-chloro

isothiocyanocarbamoyl chloride was gently refluxed for 2 hr during which evolution of HCl
was noticed. The progress of reaction was monitored by TLC. After completion of the reaction,
the reaction mixture was brought to room temperature and the solvent removed under reduced
pressure to obtain residue. This residue was triturated several times with petroleum ether (60-
80oC) to afford a pale yellow solid (3a).

3a: IR (KBr) : 3155.5 (Ar-H), 2951.0 (C-H aliphatic), 1593.2 (C=N), 1508.3 (C-C),
1131.0 (C-N), 1143.7 (C=S), 752.2 (C-S), 503.4 (S-S), cm-1; 1H NMR (δ in ppm, CDCl3): δ
7.94-7.22 (9H, m),; δ 2.358-2.353 (3H, s, CH3 ) Mass (m/z ): 316 (M+), 300, 225, 211 , Anal.
Calcd for C15H12N2S3, HCl: C, 56.96; H, 3.79; N, 8.86; S, 30.37; Found: C, 56.92; H, 3.75; N,
8.90; S, 30.34.

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On the basis of all above facts the product with m. p. 122oC was assigned the structure 3-thio-
4-phenyl-5-tolyl-[1, 2, 4]-dithiazolidine [hydrochloride]
When the reaction of N-p-tolyl-S-chloro-isothiocarbamoyl chloride was extended to several
other Ammonium phenyl dithiocarbamate corresponding 3-thio-4-aryl-5-tolyl-[1, 2, 4]-
dithiazolidine [hydrochloride] (3b-g) have been isolated.
- + R N C S
R NH2 C S NH4 S-Cl Reflux
S N C 2 hr
Cl N C S , HCl
1(a-g) CHCl3 S
2
3(a-g)
Ammonium aryl N-p-tolyl-S-chloro- 3-thio-4-aryl-5-tolyl
dithiocarbamate isothiocarbamoyl chloride -[1, 2, 4]-dithiazolidine
[hydrochloride]
Where, R= (a) Phenyl, (b) o-Cl-phenyl, (c) m-Cl-phenyl, (d) p-Cl-phenyl, (e) o-tolyl,
(f) m-tolyl (g) p-tolyl.

3c: IR (KBr) : 3032.1 (Ar-H), 2769.7 (C-H aliphatic), 1593.2 (C=N), 1541.1 (C-C),
1131.0 (C-N), 1207.4 (C=S), 715.5 (C-S), 532.3 (S-S), cm-1; 1H NMR (δ in ppm, CDCl3): δ
7.94-7.22 (8H, m),; δ 2.358-2.353 (3H, s, CH3 ) Mass (m/z ): 350 (M+), 335, 315, 259, Anal.
Calcd for C15H11N2S3Cl, HCl: C, 51.42; H, 3.14; N, 8.00; S, 27.42; Found: C, 51.46; H, 3.10;
N, 8.04; S, 27.43.

Table -1: Physical data for characterization of compounds (3a-g)

Compd Yield Rf M.P. Analysis (%): Found
0
% C (calcd)

N S
3a 78.00 0.65 122 8.90(8.86) 30.34(30.37)
3b 72.00 0.70 110 8.02(8.00) 27.38(27.42)
3c 68.00 0.50 101 8.04(8.00) 27.43(27.42)
3d 55.00 0.52 123 7.50(8.00) 27.38(27.42)
C and H analysis was found satisfactory in all cases.
Acknowledgement
Authors are thankful to SAIF, CDRI, Chandigarh for providing the spectral data
and also Dr. V. D. Nanoty for encouragement and necessary facilities.

References:
1. A. Dandia, R. Singh, S. Khaturia, C. Merienne, G. Morgant, and Loupy A, Bioorg
Med Chem., 1; 14(7), (2006), 2409-17.
2. P. R. Kawle. P. Deohate, and B. N. Berad, Int. J. of Chem. Sci., 10 (2012), 895-900.
3. R.M. Kharate, P.P. Deohate, and B.N. Berad, Int. J. Chem. Sci., 7(4), (2009), 2843.

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4. P. T. Agrawal, S. P. Deshmukh, J. Indian Chem. Soc., 86, (2009), 645-648.

5. P. Manna, R. Singh, K.K. Narang, and S.K. Manna, Indian J. Chem., 44B, (2005)1880.
6. Wilkerson W. W., Dax S., Cheatham W. W.; J. Med. Chem.; 40, (1997), 4079-4088.
7. Nefzi A., Ostresh J. M., Houghten R. A.; Chem. Rev.; 97,(1997), 449-472.
8. Z. A. Kaplancikli , G. Turan-Zitouni, A. Ozdemir, G. Revial, Eur. J. Med. Chem., 43, (2008),
155-159.
9. N. A. Al-Masoudi, Y. A. Al-Soud, M. Irene. Lagoja, Carbohy. Res., 318, (1999), 67-74.
10. A. M. Abdel-Megeed, H. M. Abdel-Rahman, Gamal-Eldien S., Eur. J. Med. Chem., 44, (2009),
117-123.
11. P. X. Franklin, A. D. Pillia, P. D. Rathod, S. Yerande, M. Nivsarkar, H. Padh, K. K. Vasu, V.
Sudarsanam, Eur. J. Med. Chem., 43, (2008), 129-134.
12. V. B. Jadhav, M. V. Kulkarni, V. P. Rasal, S. S. Biradar, M. D. Vinay, Eur. J. Med. Chem., 43,
(2008), 1721-1729.
13. R. K. Rawal, R. Tripathi, S. B. Katti, C. Pannecouque, Erik De Clercq, Eur. J. Med. Chem., 43,
(2008), 2800-2806.
14. K Ishihara, T Ichikawa, Y Komuro, S Ohara and K Hotta, Arzneim Forsch, Drug Res, 44 (1994),
827.
15. D. Havrytyuk, L. Mosula, B. Zimenkovsky, O. Vasylenko, A Gzella and R. Lesyk, Eup.
J. Med. Chem., 45(11) (2010), 5012.
16. R. M. Silverstein, G. C. Bassler and T. C. Morrill, Spectrometric Identification of Organic
Compounds”, 5th ed., John Wiley and Sons, Inc, New York, (1991).
17. N.B. Colthup, L.H. Daly and S.E. Wibereley, Introduction to Infrared and Raman spectroscopy
“Academic Press, New York, (1964).
18. D.H.Williams and L.Fleming, Spectroscopic Meyhods in Organic Chemistry”, 4th ed., Tata-
McGraw- Hill, (1988).
19. B.S. Furniss, A.J. Hannaford, W.G. Smith, A.R. Tatchell, “Vogel text book of practical
organic, chemistry”, FLBS,4th ed.; p (a) 456 (b), 649 (1980).

Gaphical Abstract
- + S-Cl
R NH2 C S NH4 N C
S Cl
1(a-g) 2
CHCl3 Reflux N-p-tolyl-S-Chloro
Ammonium aryl
2 hr isothiocarbamoyl Chloride
dithiocarbamate

R N C S
, HCl
N C S
S
3(a-g)
3-thio-4-aryl-5-tolyl-[1, 2, 4]-dithiazolidine [hydrochloride].

R= (a) Phenyl, (b) o-Cl-phenyl, (c) m-Cl-phenyl, (d) ) p-Cl-phenyl,

(e) o-tolyl, (f) m-tolyl, (g)p-tolyl
Kavita M. Heda*

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19

Characterization of Diesel Mixtures by Ultrasonic Methods

K. P. Belsare1 N. B. Selukar2
Department Of Chemical Technology,
Sant Gadge Baba Amravati University, Amravati, Maharashtra, India
Email: [email protected]

Abstract: Petroleum fuel has a higher demand in the automobile sector. To meet this demand
petroleum fuels are adulterated by some unwanted impurities. These impurities cause bad
effects on fuel engines as well as the environment. To protect the environment and automobile
engines; the detection of adulteration is important. The ultrasonic method is a recent technique
for adulteration detection so the ultrasonic velocity method is used with physiochemical
parameters for the characterization of Diesel.
Keywords: Diesel Mixture, Physiochemical characterization, ultrasonic method, Evaluation
techniques
1. Introduction
Petroleum products have a lot of demand in the current market because of increasing
automobile sectors to meet increasing demand for goods and public transport [1]. To meet this
increasing demand there are some malpractices of adding adulterant in petroleum fuel and their
presence can not be identified by simple visual inspection. But because of this adulteration, it
is dangerous to the environment, human health as well as functioning of the engine. Therefore
characterizing and determining its quality and being able to distinguish between these types is
of utmost importance to protect the consumer as well as the environment. Adulteration of fuel
becomes an acute problem today [2][3].
The ultrasonic method has recently used the Nondestructive method for the characterization
of liquid fuel. Evaluation of petroleum fuel by ultrasonic method can be a better option.
Density, viscosity, API gravity these physiochemical properties are of importance to
characterize the different types of fuels but these are not sufficient to detect adulteration in fuel
[3].
The objective of this study is to demonstrate the ability of ultrasonic methods to characterize
petroleum products with physiochemical methods for detecting adulteration easily[4].
2. Materials and Methods
Diesel purchased from HP petrol pump. I have to study the adulteration of kerosene in
diesel I purchased kerosene from the local market. The Kerosene purchased was blue. Both
samples were kept in an airtight plastic tank. These diesel and kerosene were evaluated as per
IP/ASTM norms. Then these results were compared with IS/ BIS norms for kerosene and
diesel. Following Physiochemical characterization has been done on Diesel and kerosene
samples.
 Density
 Specific Gravity
 API gravity
 Viscosity
 Ultrasonic velocity

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2.1. Preparation of custom proportions of blends of fuel and adulterants

As an adulterated sample unwanted material is present in various amounts
commonly in the range 20-80%. Hence the blends are prepared in such a way that it
will cover the adulteration range. The blends are prepared as follows:

Sr. No. % of kerosene % of Diesel

1. 20 80

2. 40 60

3. 60 40

4. 80 20

Table 1. Percentages of Blends

These blends were kept in airtight bottles.

2.2. Evaluation of blends as per IP or ASTM norms:

The blended samples were subjected to various tests as per IP/ASTM norms such as Density,
Specific Gravity, Viscosity, and ultrasonic velocity. Their tests have shown the variation in
properties will indirectly indicate the quantity of adulterant in the sample.
3. Experimental Procedure:
The experimental procedure adopted for this experiment is given below
1) Specific gravity and API Gravity is calculated by the following formula

𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑠𝑎𝑚𝑝𝑙𝑒
Specific Gravity at 25 0c =𝑊𝑒𝑖𝑔ℎ𝑡 𝑜𝑓 𝑑𝑖𝑠𝑡𝑖𝑙𝑙𝑒𝑑 𝑤𝑎𝑡𝑒𝑟 (for equal volume)

API Gravity is calculated by the formula

141.5
API = 𝑆𝑝.𝐺𝑟𝑎𝑣𝑖𝑡𝑦 𝑎𝑡 15.6 – 131.5 (Sp. Gravity is calculated at 15.60 c)

2) Viscosity: The viscosity of the sample has been calculated by U – U-tube Viscometer.
3) Ultrasonic velocity measurement: Ultrasonic velocity has been measured by using
Ultrasonic Interferometer. The formula used for measuring ultrasonic velocity is given
below
Velocity = Wavelength × Frequency
Where velocity is measured for different blends. And Observations are given in Table 2.

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3.1. Experimental Observations

Diesel with kerosene is evaluated as follows: All tests were done at 250 c.
Sample 100% D + 80 % D + 60 % D 40% D + 20% D + 0 % D
parameter 0% K 20 %K + 40 % K 60 % K 80 % K +100 % K
Density 0.8322 0.8223 0.8222 0.8208 0.8181 0.7800
Specific 0.8336 0.8288 0.8187 0.8173 0.8146 0.7811
Gravity
API 40.06 41.3140 41.3172 41.6289 42.1882 43.1992
Gravity
Viscosity 4.35 2.93 2.73 2.59 2.47 2.38
Ultrasonic 1325 1315.2 1281.6 1278.08 1277.4 1267.5
velocity

Table 2. Experimental Observation of Blends of Kerosene with Diesel

4. Results and Discussion
From the above experimental procedure, it is observed that, For the Kerosene diesel adulterated
sample, as
A. As the percentage of adulterants increases density of diesel decreases.
B. As the percentage of kerosene increases in diesel, specific gravity also decreases.
C. As the percentages of kerosene increase API gravity increases.
D. viscosity decreases as the adulterant increases.
E. Ultrasonic velocity decreases as percentages of kerosene in diesel increases.

References
1. Sh. R. Yadav, K. Murthy V, D. Mishra, and B. Baral, “Estimation of petrol and diesel
adulteration with kerosene and assessment of the usefulness of selected automobile fuel
quality test parameters”, International Journal of Environmental Science and Technology,
Vol.1, No. 4, pp. 253-255, (2005)
2. A. Jimenez, M.Rufdo, J.Panigua, A, Gonzalez-Mohino, L-S.Olegario, “New findings of
edible oil characterization by ultrasonic parameters”, Food Chemistry, vol. 374, pp 1-6,
(2022)
3. G. M. Ramteke , L.B.Revatkar , R. V. Phadke , N.L.Chutke, “ Analysis of Petrol : A
Clarification for Purity of Petrol” , ESR Journal , pp. 1-8.(2016)
4. B. Alouache , F. K. Khechena, F. Lecheb , T. Boutkedjirt, “ Characterization of Olive oil
by Ultrasonic and Physiochemical Methods” , Elsevier, 70, 1875-3892 (2015).

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20
Phytochemical and Proximate Analysis of Leaves of Heteropogon
Contortous (L.) Beauv
Miss Mosim Raza H. Shaikh
Jagadamba Mahavidyalaya, Achalpur
[email protected]
ABSTRACT :
Recently Proximate analysis and phytochemical analysis of Heteropogon Contortous (L.)
Beauv from Wazzar Village,located in Achalpur tehsil of Amravati,Maharashtra.The leaves
sample contained tannin, saponin, protein, steriods, terpenoids, carbohydrate,alkaloids,
flavonoids.Proximate analysis of moisture, ash,crude fiber,crude Protein, solubility were
check. The values of it is moisture (27.78%), cold water (64.4%), hot water (50.4%), 1%NaOH
(51.76%), 1%HCl (54.57%), ash content (13.48%). These results indicate that the Heteropogon
Contortous (L.) Beauv. contains nutrients elements that will be useful in nutrition. Also the
existence of some phytochemicals like tannin, saponin and steroids illustrated medicinal action
of the plant in its therapeutic uses. The result of their phytochemcial screening could justify
the observed activities and validate their use in herbal medicine.
Keywords: Proximate composition, Phytochemical analysis, Heteropogon Contortous (L.)
Beauv
INTRODUCTION :
The term “medicinal plants” includes various type of plants used in herbalism and some of
these plants have medicinal activities. Medicinal plants are the “backbone” of traditional
medicine, which means more than 3.3 billion people in the less developed countries utilize
medicinal plants on a regular basis [1].

Medicinal plants are an integral component of research developments in the Pharmaceutical

industry. Such research focuses on the isolation and direct use of active medicinal constituents,
or in the development of semi-synthetic drugs, or still again on the active screening of natural
products to yield synthetic pharmacologically-active compounds. Natural products play an
important role in drug discovery process including the provision of basic compounds affording
less toxic and more effective drug molecules, serve as extremely useful natural drugs,
exploration of biologically active prototypes towards newer and better synthetic drugs and
modification of inactive natural products by suitable biological or chemical means into potent
drugs [2].

Spear grass (Heteropogon contortus (L.) Beauv. ex Roem. & Schult.) Is a tropical perennial
grass. It grows to a height of 50 to 150 cm, is tufted and highly variable. Its stems are
geniculated at the base, erect at their upper levels, often branched, particularly at flowering [3].
The leaves are green or bluish green, usually glabrous or with few long hairs at the base. The
leaf-blade is folded when young, and then flat at maturity, 3-30 cm long, 2-8 mm broad, and
somewhat canoe-shaped at the apex [3, 4]. The inflorescence is a 3 to 8 cm long raceme borne
single or in pairs at the axil of the upper leaves. The spikelets are paired and very dissimilar
according to their position on the raceme. Male or sterile spikelets are awnless, sessile and
borne at the base of the raceme, or pedicellate and borne at the apex. Bisexual spikelets are
only borne at the apex and they are all awned. The long awns (5-10 cm long) and the way they
become twisted as the seeds mature are a characteristic trait of spear grass. The seed is a

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caryopsis, 3.5-4.5 mm long, grooved and whitish in colour [3, 4, 5]. There were considerable
numbers of local species and varieties in the early botanical literature. Only a few commercial
varieties are available, for example "Rocker" from Arizona and "Kahoolawe" from Hawaii
[4].Aside from phytoconstituent identification, it is also critical to analyze plants for their
proximate and mineral compositions to expand knowledge of their nutritional health benefits.
The review of the literature revealed that no previous work on the proximate analysis of
Heteropogon Contortous (L.) Beauv. had been conducted.

MATERIALS AND METHODS

Collection of Samples
The leaves of the plant Heteropogon contortus (L) Beauv was collected from Wazzar
Village,located in Achalpur tehsil of Amravati,Maharashtra., The whole plant was gently
uprooted from the ground and then the leaves separated from the root after which they were
dried at ambient temperature. The dried leaves were milled to fine powder, thereafter it was
stored in an airtight container in the prior to its use.

Proximate analysis:
As per Aurveda, herbal drugs in the form of oral dosage are given to the patient as powdered
materials mixed with water. For that, we performed proximate analysis to determine the
physicochemical properties as well as solubility and nutrient content of herbal drug materials.
Using a standard procedure and formula from the reported literature, the moisture content, ash
content, crude protein, crude fiber, crude fat were determined.[ 6-8]
Moisture, ash and solubility were determined using the Association of official analytical
chemists methods [5]. Crude fiber was estimated from the loss in weight on ignition of dried
residue following digestion of fat-free samples.

Ash content:
An ignited and weighed silica crucible which contains 1gm of the powdered sample was
incinerated slowly by raising the temperature in a muffle furnace at 450ºC for 4 – 5 hours. It
was cooled in desiccators and weighed. The procedure was repeated until a constant weight
was obtained. The percentage of total ash was calculated.

Moisture Content:
The oven-drying method was used to determine moisture. The weight of the empty crucible
was recorded and kept in the oven for 1 hour at 1100C, then 1 gm of dried sample was placed
in a crucible (W1) and kept in the oven for 1 hour at 1100C. It was then cooled and weighed
until the weight remained constant (W2). The moisture content (%) was calculated.

Crude fiber content:

After extraction with petroleum ether, 5 g of the dry material was boiled with 2% H2SO4 and
NaOH solution for 30 minutes, respectively. It was filtered and washed the residue with boiling
water and dried for 2 hrs at 130o C, cooled in a desiccator and weighed. Then the residue was
ignited at 550 °C for 25 min. and cooled in a desiccator before reweighing.

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Table -1
Sr.No. Solubility in Actual weight of Loss of wt. of %
sample taken in gm sample in gm
1 Cold water 1.000 0.644 64.4
2 Hot water 1.000 0.504 50.4
3 Dil.NaOH 1.000 0.5176 51.76
4 Dil.HCl 1.000 0.5457 54.57
5 Ash Content 1.000 0.1348 13.48
6 Moisture 1.000 0.2778 27.78
Content
7 Crude Fiber --- --- 33.7
8 Crude --- --- 2.7
Protein

Phytochemical analysis :
Phytochemical analysis of the Heteropogon contortus (L) Beauv leaves.

S.No. Content Methanol Aqueous

1 Amino Acids - -
2 Proteins - -
3 Carbohydrates + +
4 Saponins - +
5 Alkaloids + +
6 Phenols + +
7 Steroids + +
8 Terpenoids + +
9 Cardiac Glycosides + +
10 Flavonoids + +
11 Tannins + +
12 Anthroquinones + +

“+” = present, “-” = absent

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RESULT AND DISCUSSIONS

The results of the proximate analysis support the use of the leaves as a food supplement. From
the result, it was found that the total ash content obtained from dry leaves is 13%. Ash content
is generally taken to be a measure of the mineral content of original food.The moderate
moisture content provides for an activity of water-soluble enzymes and coenzymes needed for
the metabolic activities of the plant. The total moisture content is 27.78 % .The proximate
composition of the plant is depicted in table no.1.

The presence of flavonoids inferred that the leaves has the biological functions like
antioxidant,allergies protection, free radical, inflammation, ulcers, hepatotoxins, tumor and
viruses[8]. Flavonoids are water soluble free radical and antioxidants which prevent oxidative
cell damage, and have strong anti-ulcer and anticancer activity[10]. Saponin content suggest
that usefulness of the leaves as a productivity agent. The saponin level is low, either compared
with the results from another works. Alkaloids are the most efficient medicinally
significant bioactive substances in plants. Alkaloids and the synthetic derivatives are used as
medicinal agents because of their bactericidal and analgesic properties. Tannins healing of
wounds and inflamed mucous membranes. This are water soluble phenolic compounds which
precipitate proteins. They exist in all plants .Tannins add to proteins making them bio-
unavailable [11]

CONCLUSION
This type of study will be applicable for the pharmaceutical, medicinal, agricultural, industrial
and biochemical sciences. This study also shown that proximate, phytochemical analysi of
Heteropogon contortus (L) Beauv leaves is a balanced and rich source of macro- and
micronutrients. So the further study will be carry out on this plant.

REFERENCES
1. Singh, R. (2015). Medicinal plants: A review. Journal of Plant Sciences, 3(1-1), 50-55.
2. Saad, R., Pohyeen, T., Khan, J., Wenji, L., Sultan, S., & Abdul Hameed, J. (2014). Phytochemical screening
and antioxidant activity of different parts from five Malaysian herbs. International journal of science and
technology, 19(2), 1336-1347.
3. Cook, B. G., Pengelly, B. C., Brown, S. D., Donnelly, J. L., Eagles, D. A., Franco, M. A., ... & Schultze-Kraft,
R. (2005). Tropical Forages: an interactive selection tool. Tropical Forages: an interactive selection tool.
4. Soromessa, T. (2011). Heteropogon contortus (L.) P.Beauv. ex Roem. & Schult.. Record from Protabase. Brink,
M. & Achigan-Dako, E.G. (Editors). PROTA (Plant Resources of Tropical Africa / Ressources végétales de
l’Afrique tropicale), Wageningen, Netherlands
5. Food and Agricultural Organization (FOA), (2012). Grassland Index. A searchable catalogue of grass and
forage legumes. FAO, Rome, Italy
6. Dhivya, R., & Manimegalai, K. (2013). Preliminary phytochemical screening and GC-MS profiling of ethanolic
flower extract of Calotropis gigantea Linn.(Apocynaceae). Journal of Pharmacognosy and Phytochemistry, 2(3),
28-32.

7. P. K. Mukherjee, Elsvier, First Edition, 2019, 237-328, Quality Control Evaluation Herbal Drugs, doi:
10.1016/B978-012-813374-3.00007-7.
8. P. K. Mukherjee, Business Horizon Pharmaceutical Publications, New Delhi, 2008, 187-191.
9. AOAC, 20th edition. Rockville, Maryland, USA, 2016.
10.Akindahunsi A.A. & Salawu S.O., African J. Biotech., 4, 2005, 497-501.
11. Sofora L.A., Medicinal plants and traditional medicine in Africa. (Spectrum books Ltd.)1993, p.55-71.

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21
A Review of Computer-Aided Drug Discovery (CADD)
Manojkumar O. Malpani
Associate Professor
Department of Chemistry
Shankarlal Khandelwal Arts, Science and Commerce College, Akola, India
Email Id: [email protected]

ABSTRACT
Computer-aided drug discovery (CADD) is an innovative approach that leverages
computational tools to accelerate and improve the drug development process. With the
advancement of technology, CADD has become an increasingly prevalent tool in the
pharmaceutical industry. By simulating and analyzing molecular interactions, predicting drug-
target binding affinities, and screening vast libraries of compounds, CADD has proven to be
instrumental in identifying potential lead compounds with therapeutic potential. Through its
ability to efficiently sift through enormous amounts of data and generate accurate predictions,
CADD enables researchers to prioritize the most promising candidates for further investigation,
ultimately streamlining the drug discovery timeline and increasing the chances of successfully
developing effective treatments for various diseases.
Keywords: CADD, QSAR, Molecular docking.

1. INTRODUCTION
Drug discovery is a complex and time-consuming process that involves the
identification and development of new therapeutic molecules to treat various diseases.
Traditionally, this process heavily relied on experimental methods, which can be expensive,
time-consuming, and limited in their ability to explore vast chemical space. However, with the
advent of computational chemistry, a powerful field at the intersection of chemistry, biology,
and computer science, drug discovery has witnessed significant advancements. Computational
chemistry employs computer simulations, algorithms, and mathematical models to accelerate
the drug discovery process, offering valuable insights into the behaviour of molecules and their
interactions with biological targets. This review provides an overview of the role of
computational chemistry in drug discovery, explores the techniques and tools employed,
discusses the challenges and limitations, and highlights the future perspectives and
advancements in this exciting field.
Overview of the drug discovery process
We are probably wondering how new drugs are discovered. It's not as simple as mixing
random chemicals in a lab and hoping for the best. No, it's a carefully orchestrated process that
involves a lot of trial and error, and a pinch of luck. The drug discovery process typically starts
with identifying a target, which could be a receptor or an enzyme involved in a disease pathway.
Once the target is identified, scientists embark on a mission to find a molecule that can interact
with this target and either enhance or inhibit its function.
Now, you're probably thinking, "Well, that sounds like a lot of work!" And you're right!
But fear not, because this is where computational chemistry swoops in to save the day.

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Computational chemistry is like the superhero of drug discovery, using powerful computer
modelling techniques to predict the behaviour of molecules and their interactions with targets.
2. ROLE OF COMPUTATIONAL CHEMISTRY IN DRUG DISCOVERY
Computational chemistry brings a whole host of advantages to the drug discovery table.
For one, it saves time and money. Instead of blindly testing thousands of molecules in the lab,
scientists can use computational methods to screen and prioritize the most promising
candidates. It's like having a crystal ball that tells you which molecules are worth pursuing.
Another advantage is that computational chemistry allows scientists to explore a vast chemical
space. They can virtually generate and test millions of compounds without ever stepping foot
in a lab. It's like having a magical laboratory that's not constrained by physical limitations.
Integration of Experimental and Computational Approaches
Now, you might be thinking, "Well, if computational chemistry is so amazing, can we
just replace all the lab work with computers?" Not quite. While computational methods are
incredibly powerful, they're not infallible. They still need to be validated with experimental
data to ensure accuracy. That's why scientists often employ an integrated approach, combining
both computational and experimental techniques. By synergizing the powers of computational
chemistry and good old-fashioned test tubes, they can accelerate the drug discovery process
and increase the chances of success.

3. TECHNIQUES AND TOOLS IN COMPUTATIONAL CHEMISTRY FOR DRUG

DISCOVERY
Structure-Based Drug Design
Structure-based drug design is like playing molecular Tetris. Scientists use the three-
dimensional structure of the target protein to design molecules that fit into specific pockets or
binding sites. It's all about finding the perfect shape and electrostatic properties that will allow
the molecule to interact with the target desirably.
Ligand-Based Drug Design
Ligand-based drug design takes a different approach. Instead of focusing on the target
protein, scientists analyse molecules that have already shown activity against the target. They
then extract common features and create new molecules based on those patterns. It's like
reverse engineering a recipe to create a delicious new dish.
Quantitative Structure-Activity Relationship (QSAR)
Ever wondered if there's a mathematical equation for drug discovery? Well, QSAR
comes pretty close. It's a technique that uses statistical models to establish correlations between
the structure of a molecule and its biological activity. By understanding these relationships,
scientists can predict the activity of new molecules and prioritize those with the highest chances
of success.
4. VIRTUAL SCREENING AND MOLECULAR DOCKING IN DRUG DISCOVERY
Virtual screening is like speed dating for molecules. It involves screening large libraries
of compounds against a target protein to identify potential hits. Instead of physically testing
each molecule, scientists use computer algorithms to predict how well they will interact with

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the target. It's a quick and efficient way to narrow down the options and focus on the most
promising candidates.

Molecular Docking Methods

Molecular docking is like playing a game of molecular lock and key. Scientists use
computational algorithms to predict how a small molecule (the ligand) will bind to a target
protein (the receptor). By exploring different orientations and conformations, they can
determine the most favourable binding pose. It's like solving a puzzle where the reward is a
potential new drug.
Validation and Analysis of Docking Results
Of course, docking results are not to be taken at face value. They need to be validated
and analysed to ensure their reliability and relevance. Scientists use various techniques, such
as scoring functions and experimental validation, to assess the quality of the docking results.
It's like a reality check for those molecules that thought they were the perfect match. And there
you have it; a whirlwind tour of how computational chemistry shakes things up in the world of
drug discovery. It's a fascinating field that combines science, technology, and a dash of
creativity to create new medicines that can change lives. So next time you hear about a
groundbreaking drug, you can appreciate the superheroes behind the scenes, harnessing the
power of computers to make it happen.
5. MOLECULAR DYNAMICS SIMULATIONS AND QUANTUM MECHANICS IN
DRUG DISCOVERY
Role of Molecular Dynamics Simulations
In the world of drug discovery, scientists are turning to computational chemistry
techniques such as molecular dynamics simulations to understand the behavior of molecules at
the atomic level. By simulating the movements and interactions of molecules over time,
researchers can gain valuable insights into how drugs interact with their target proteins. This
knowledge is crucial for designing and optimizing drug candidates with higher efficacy and
fewer side effects.
Quantum Mechanics Applications in Drug Discovery
Quantum mechanics, on the other hand, provides a more detailed and accurate
description of molecular properties and interactions. It allows scientists to calculate properties
such as molecular energies, electronic structures, and reaction mechanisms. This information
is vital in understanding the fundamental aspects of drug action and can guide the development
of more potent and selective drugs.
Combining Molecular Dynamics and Quantum Mechanics
To achieve even greater accuracy, researchers often combine molecular dynamics
simulations with quantum mechanics calculations. This hybrid approach, known as QM/MM
(quantum mechanics/molecular mechanics), takes advantage of the strengths of both methods.
It allows for a more realistic and comprehensive representation of the drug-target interactions,
taking into account the dynamic nature of the system and the quantum effects that are critical
for understanding complex biological processes.
6. CHALLENGES AND LIMITATIONS OF COMPUTATIONAL CHEMISTRY IN
DRUG DISCOVERY

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Accuracy and Reliability of Computational Models

Despite the significant advancements in computational chemistry, there are still
challenges when it comes to the accuracy and reliability of the models used in drug discovery.
The accuracy of predictions heavily relies on the quality of the force fields, parameterization
methods, and approximations employed. It is essential for scientists to constantly refine and
validate these models against experimental data to ensure their reliability.
Handling Large Data Sets and Computational Complexity
Another challenge is the exponential growth of data in drug discovery. With the
increasing availability of genomic, proteomic, and chemical data, computational chemists must
develop efficient algorithms and techniques to handle and analyze these large data sets.
Additionally, the computational complexity of modeling drug-target interactions requires
substantial computational resources, which can be a limiting factor for many research groups.
Limitations in Predicting Drug Efficacy and Safety
Computational chemistry methods have their limitations in accurately predicting drug
efficacy and safety. While they can provide valuable insights into drug-target interactions, they
cannot capture the complexities of the human body fully. Factors such as metabolism,
pharmacokinetics, and off-target effects still require experimental validation. Therefore,
computational chemistry should be considered as a complementary tool rather than a
standalone solution in drug discovery.
7. FUTURE PERSPECTIVES AND ADVANCEMENTS IN COMPUTATIONAL
CHEMISTRY FOR DRUG DISCOVERY
Emerging Technologies in Computational Chemistry
Exciting advancements in computational chemistry are on the horizon. Techniques such
as accelerated molecular dynamics, enhanced sampling methods, and machine learning-based
approaches are actively being developed to overcome the limitations of current methodologies.
These emerging technologies hold promise for more accurate predictions and deeper insights
into drug-target interactions.
Integration of Artificial Intelligence and Machine Learning
Artificial intelligence (AI) and machine learning (ML) are revolutionizing the field of
drug discovery. By leveraging vast amounts of data, AI and ML algorithms can identify
patterns, predict drug-target interactions, and even suggest novel drug candidates. The
integration of these technologies with computational chemistry has the potential to
significantly accelerate the drug discovery process and increase the success rate of developing
new medications.
Potential Impact on Drug Discovery Process
The advancements in computational chemistry are poised to reshape the drug discovery
landscape. By enabling faster and more accurate predictions, computational chemistry can help
researchers prioritize potential drug candidates, optimize their properties, and reduce the need
for extensive experimental screening. Ultimately, this can lead to a more efficient and cost-
effective drug discovery process, benefiting patients worldwide by providing faster access to
safer and more effective medications. In conclusion, computational chemistry has
revolutionized the field of drug discovery, enabling scientists to expedite the identification and
development of potential therapeutics with greater precision and efficiency. The integration of

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computational methods with experimental approaches has proven to be a powerful

combination, offering valuable insights into molecular interactions, structure-activity
relationships, and drug efficacy. However, challenges such as accuracy, computational
complexity, and the limitations of predictive capabilities still exist. Nonetheless, with the
continuous advancements in technology, the future of computational chemistry in drug
discovery holds immense promise. As researchers explore emerging technologies, incorporate
artificial intelligence, and refine computational models, we can expect even greater strides in
the discovery and development of novel drugs that improve the health and well-being of
individuals worldwide.
CONCLUSION
The future of computational chemistry in drug discovery is promising. Emerging technologies
such as artificial intelligence and machine learning are being integrated into computational
chemistry workflows, allowing for more efficient and accurate predictions. Continued
advancements in computational power and algorithms will further enhance the capabilities of
computational chemistry. These advancements have the potential to revolutionize the drug
discovery process, leading to the development of more effective and personalized therapeutics
to address the unmet medical needs of patients.

REFERENCES:
1. Surabhi and B. K. Singh, COMPUTER AIDED DRUG DESIGN: AN OVERVIEW.
Journal of Drug Delivery & Therapeutics. 2018; 8(5):504-509.
http://dx.doi.org/10.22270/jddt.v8i5.1894.
2. J. J. Naikwadi, N. Desai, S. V. Patil, Computer aided drug design- an overview. IJCRT,
Volume 9, Issue 10, October 2021,ISSN: 2320-2882.
3. K. S. Dhamal, S. A. Waghmare, H. Kamble, et.al., Role Of Computer Aided Drug
Design In Drug Discovery & Development. International Journal of Research in
Engineering and Science (IJRES). Volume 10, Issue 3, 2022, PP. 01-05.
4. I. Hoque, A. Chatterjee, S. Bhattacharya and R. Biswas, An Approach of Computer-
Aided Drug Design (CADD) Tools for In Silico Pharmaceutical Drug Design and
Development. Int. J. Adv. Res. Biol. Sci. (2017). 4(2): 60-71.
http://dx.doi.org/10.22192/ijarbs.2017.04.02.009.
5. R. S. Waghmode, A. B. Lasure, S. B. Bavage, N. B. Bavage, Computer Aided Drug
Design (CADD). IJIRT, Volume 8, Issue 3, ISSN: 2349-6002.

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22

Synthesis and Anti-microbial Activity of Novel 3-[(3-substitutedamino-

1,2,4-thiadiaz-3-yl)]amino-5-N-TAG-amino-1,2,4-thiadiazole [11b]

M. R. Raghuvanshi
Department of Chemistry, Arvindbabu Deshmukh Mahavidyalaya, Bharsingi, Dist-Nagpur, Maharashtra,
India-441305 Corresponding [email protected]

Abstract
In this study new 3-[(3-substituted amino -1,2,4-thiadiaz-3-yl)] amino-5-TAG-amino-1,2,4-
thiadiazole [11] were synthesized by the oxidative cyclization of 1-[S-TAG-N-
substitutedthioamido] dicyanadiamide [8] and 1-[(N-TAG) thioamido]-5-
substitutedformamidino-2-imino-4-thiobiuret [9] respectively in chloroform medium using
liquid bromine as oxidizing agent. The newly synthesized compounds isolated in these reaction
were characterized and justified on the basis of conventional elemental analysis, chemical
characteristics, IR, NMR and Mass spectral analysis. The synthesized compounds were
screened for antimicrobial activities by disc diffusion method.

Keywords – Synthesis, Dicyandiamide,Thiadiazol, Antimicrobial activity

Introduction
During last few decades, heterocyclic molecules containing nitrogen, sulfur and oxygen (five
and six membered rings) have become the area of interest for medicinal chemists and
researchers owing to their wide spectrum of biological/pharmacological activities.1. Thiadiazol,
a five membered heterocyclic moiety containing one sulfur and two nitrogen atoms existing in
four different isomeric forms, 1,2,3-thiadiazole; 1,2,4-thiadiazole and 1,2,5-thiadiazole and
1,3,4-thiadiazole has established its significant importance due to comprehensive
pharmacological activities.2-4. 1,2,4-Thiadiazoles are heteroatom rich heterocycles with a vital
role in medicinal chemistry programmes; many clinically approved pharmaceuticals contain
this heterocycle.5
1,2,4-thiadiazoles are considered as most significant subclass of bioactive five-membered
organic compounds for medicinal chemistry6and showed a remarkable biological activities
such as cyclooxygenase inhibitors7, human leukemia8, antibacterial9, antiulcerative10,
antihypertensive11, cathepsin B inhibitors12, anticonvulsant13, antidiabetic14, anti-
inflammatory7, and allosteric modulators15.
The wide range of application of 1,2,4- thiadiazols as pharmacophores in medicinal chemistry
has attracted great interest in their synthesis.16
As a part of research work presently been undertaken in this laboratory for the synthesis of
heteroacycles and heterocycles, it was thought interesting to investigate the oxidative
cyclization of 1-[(N-TAG) thioamido]-5-substitutedformamidino-2-imino-4-thiabiuret [9]
with liquid bromine in chloroform medium to obtain a novel series of 3-[(3-substitutedamino
-1,2,4thiadiaz-3-yl)] amino-5-N-TAG-amino-1,2,4-thiadiazole [11] In this study, a new series
of 1,2,4-thiadiazole derivatives (11a–e) were synthesized in good yields, and the structures of
the compounds were confirmed by IR, 1 H-NMR, and elemental analysis. For that The newly
synthesized substituted derivatives of thiocarbamide were screened for its antibacterial
activities against some gram positive & gram negative pathogens. The compounds showed
good & moderate activity against the pathogens.

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Experimental
The melting points of all the synthesized compounds were recorded using hot paraffin bath and
are uncorrected, The carbon and hydrogen analysis was carried out on Carlo-Ebra-1106
analyzer, nitrogen estimation was carried out on ColmanN-analyser-29. IR spectra were
recorded on Perkin-Elmer spectrometer in the range 4000-400 cm-1 in KBr pellets. PMR
spectra were recorded on Bruker AC-300F spectrometer with TMS as internal standard using
CDC13 and DMSO-d6 as solvent. The FAB mass spectra were recorded on a Joel SX 102/Da-
600 mass spectrometer/ Data System using Argon. The accelerating voltage was 10kV and
spectra were recorded at room temperature by using m-nitro benzyl alcohol as a matrix. The
purity of the compounds was checked on Silica Gel-G plates by TLC with layer thickness of
0.3 mm. All chemicals used were of AR grade.
Synthesis of 3-[(3-phenylamino-1,2,4-thiadiaz-3-yl)]amino-5-N-TAG-amino-1,2,4-
thiadiazole [11b]
In china dish a paste of 1-[N-TAG thioamido]-5-phenylthio formamidino-2-imino-4-
thiobiuret [9b] was prepared in chloroform. To this liquid bromine in chloroform was added
with constant stirring. Initially the colour of bromine disappeared, the addition of bromine
was continued till the colour of bromine persisted to the reaction mixture. The reaction
mixture was allowed to stand for 8 Hrs. at room conditions. It is dark brown in colour. After
the basification of the reaction mixture afforded brown coloured product, which on
crystallization gave brown crystals of [11b] yield 61%, m. p.1470C.
The probable reaction mechanism of the formation of [11b] may be stated as follows.
Scheme

Similarly 3-[(3-Amino-1,2,4-thiadiaz-3-yl)]-amino-5-N-TAG amino-1,2,4-thiadiazole

[11a], 3-[(3-methylamino-1,2,4-thiadiaz-3-yl)]-amino-5-N-TAG amino-1,2,4-thiadiazole
[11c], 3-[(3-ethylamino-1,2,4-thiadiaz-3-yl)]-amino-5-N-TAGamino-1,2,4-thiadiazole [11d],
3-[(3-allylamino-1,2,4-thiadiaz-3-yl)]-amino-5-N-TAG amino-1,2,4-thiadiazole [11e] were
synthesized from 1-[(N-TAG)thioamidino]-5-formamidino-2-imino-4-thiobiuret [9a], 1-[(N-
TAG)thioamidino]-5-methylformamidino-2-imino-4-thiobiuret [9c], 1-[(N-
TAG)thioamidino]-5-ethylformamidino-2-imino-4-thiobiuret [9d], 1-[(N-TAG)thioamidino]-
5-allylformamidino-2-imino-4-thiobiuret [9e] in bromine in chloroform medium respectively

Result ad Discussion-
Spectral Evaluation-
The IR spectrum of compound 3-[(3-phenylamino-1,2,4-thiadiaz-3-yl)]amino-5-N-TAG-
amino-1,2,4-thiadiazole was carried out in KBr pellets and is reproduce on plate No. IR-1.
The IR spectrum clearly indicated the bands due to ν-NH, ν-C-H(Ar), ν-C=O, ν-C=N, ν-RC-
N, ν-RC-S and an important absorption can be correlated in Table No. 1.

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Table No.-1

Absorption Assignment Observed Absorption expected (Cm-1)

3374.5 -NH- stretching 3500-3100
1695.8 -C=O stretching 1900-1600
1585.0 -C=N stretching 1789-1471
1213.0 -C-N Stretching 1200-1000
757.4 -C-S Stretching 800-600

The PMR spectrum of compound 3-[(3-phenylamino-1,2,4-thiadiaz-3-yl)]amino-5-N-TAG-

amino-1,2,4-thiadiazole was carried out in DMSO-d6 and CDCl3 and shown in figure 2. This
spectrum distinctly displayed the signals due to –NH protons at δ 9.5-8.6 ppm, Ar. Protons at
δ 7.77 - 7.01 ppm.
The FAB mass spectrum of 3-[(3-phenylamino-1,2,4-thiadiaz-3-yl)]amino-5-N-TAG-amino-
1,2,4-thiadiazole [11b]was recorded at room temperature by using meta nitrobenzyl alcohol as
the matrix m+ peak as well as other fragment peaks and the probable fragmentation pattern of
the molecular ion.
Antimicrobial Activity
All the synthesized compounds were screened for antimicrobial activity using disc diffusion
method17 and all the pathogen tested during analysis are human pathogens. For this Whatman
filter paper No. 1 disks of 5mm diameter were sterilized in autoclave and soaked in sample
solution, blotted on sterile filter paper. 0.1ml of the inoculums of test organism was spread
using sterile glass spreader on the surface of nutrient agar. The filter paper disks soaked in
Gentamycin (20µg/ml) (Glaxo India Ltd.) and Ciprofloxacin (20µg/ml) (Glaxo smith kline)
were used as positive controls and the filter paper disc soaked in dimethyl sulphoxide (DMSO)
were used as a solvent. The compounds were taken at a concentration or 1mg/ml using dimethyl
sulphoxide as a solvent .Fluconazole (20ug/ml)as a standard for antifungal activity. The
inhibition zones were measured in millimeter by the end of the incubation period (24 hrs. at
370C for bacteria).
Table No-2

Compounds Inhibition zone in mm

S.Typhi E.coli P. S.aurues A.nigar c.albicanes
aerogenosa
9a 8 8 -ve -ve 9 -ve
9b 10 9 -ve 8 12 10
9c 9 9 7 7 10 10
9d 9 8 9 9 9 9
9e 10 9 10 9 12 12
DMSO - - - - - -
Fluconazole - - - - 12 12
Ciprofloxacin 10 9 10 9 - -

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From the results it is clear that compounds showed remarkable and considerable antimicrobial
activity against organism. The activity of compounds were tested against all the pathogen by
disc diffusion method. The antimicrobial activity of different compound against micro-
organisms were examined in the presence study and their potency, were assessed by the
presence or absence inhibition zone and zone diameter. The result are given in Table No. 2.
From this table clearly indicates that, all the compounds are active against S. typhi, E.coli &A.
nigar but some compounds 9a and 9b are inactive against P.aerogenosa, S.aurues & c.albicans.
Conclusion-
This paper reports the synthesis of 3-[(3-phenylamino-1,2,4-thiadiaz-3-yl)]amino-5-N-TAG-
amino-1,2,4-thiadiazole [11b] & 3-[(3-Substitutedamino-1,2,4-thiadiaz-3-yl)]amino-5-N-
TAG-amino-1,2,4-thiadiazole (11a-e) derivatives and characterization by FT-IR, PMR and
mass spectrograph indicating formation of the desired product. The compounds were studied
to evaluate in vitro antibacterial and antifungal properties by disc diffusion method. The
antimicrobial activity of different compound against micro-organisms were examined in the
presence study and their potency, were assessed by the presence or absence inhibition zone and
zone diameter. From results, most of these compounds found to be potent antibacterial and
antifungal agent exhibited comparable antibacterial and antifungal activity than the standard.
It means that the compounds showed remarkable and considerable antimicrobial activity.
Acknowledgement: The author acknowledges the help of SAIF, CDRI, Lucknow for providing
the spectral data. Authors are thankful to the Department of Chemistry, Mahata Fule College of
Science Warud for providing the necessary facilities to carry out the research work & Dr.Rahul Bhagat
(Biotechnolgy) for his help in doing antimicrobial activity.
References
1. Yang Hu, Cui-Yun Li, Xiao-Ming Wang, Yong-Hua Yang, Hai-Liang Zhu., Chem. Rev. 2014,
114(10),pp5572–5610, https://doi.org/10.1021/cr400131u
2. V. Raj, A. Rai, S. Saha., Anti-Cancer Agents Med Chem, 2017,17(4),pp.500-523,
10.2174/1871520616666161013150151,PubMed: 27745547,View article View in Scopus Google Scholar
3. Serban.,Molecules,2020, 25 (4),pp.942, PubMed: 32093125. Pub Med Central: PMC7070519, View article
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10.1021/cr400131u, View article View in Scopus Google Scholar
5. Daniel G. Anstis a, Emma K. Davison, Jonathan Sperry .,Tetrahedron,Volume 150, 15 January 2024, 133767,
https://doi.org/10.1016/j.tet.2023.133767
6. Kumar, D.; Kumar, N.-M.; Chang, K.-H.; Shah, K.: Synthesis and anticancer activity of 5-(3-indolyl)-1,3,4-
thiadiazoles. Eur. J. Med. Chem. ,2010,45, pp 4664–4668
7. Unangst, P.C.; Shrum, G.P.; Connor, D.T.; Dyer, R.D.; Schrier, D.J.: Novel 1,2,4-oxadiazoles and 1,2,4-
thiadiazoles as dual 5-lipoxygenase and cyclooxygenase inhibitors. J. Med. Chem. 1992,35, pp. 3691–3698
8. Romagnoli, R.; Baraldi, P.G.; Carrion, M.D.; Cruz-Lopez, O.; Preti, D.; Tabrizi, M.A.; Fruttarolo, F.;
Heilmann, F.; Bermejo, J.; Estevez, F.,Bioorg. Med. Chem. Lett. ,2007,17, pp 2844–2848
9. Harai, R.; Sakamoto, K.; Hisamichi, H.; Nagano, N.: Structure–activity relationships of cephalosporins
having a (dimethyl isoxazolidi nio) vinyl moiety at their 3-position. J. Antibiot. ,1996,49, pp. 1162–1171
10. Kharimian, K.; Tam, T.F.; Leung-Toung, R.C.; Li, W.: Thiadiazole compounds useful as inhibitors of hb/kb
atpase. PCT Int. Appl. WO9951584A1 ,1999
11. Kohara, Y.; Kubo, K.; Imamiya, E.; Wada, T.; Inada, Y.; Naka, T.,J. Med. Chem. ,1996,39, pp. 5228–5235
12. Leung-Toung, R.; Wodzinska, J.; Li, W.; Lowrie, J.; Kukreja, R.; Desilets, D.; KarimianK.; Tam. ,Bioorg.
Med. Chem. ,2003,11, 5529–5537 (2003)
13. Castro, A.; Castano, T.; Encinas, A.; Porcal, W.; Gil, C., Bioorg. Med. Chem. 14, 1644–1652 (2006)
14. Johnstone, C.; Mckerrecher, D.; Pike, K.G.; Waring, M.J., PCT Int. Appl.WO2005121110A1 ,2005.
15. van den Nieuwendijk, A.M.C.H.; Pietra, D.; Heitman, L.; Goblyos, A.; Ijzerman, A.P., J. Med. Chem.
,2004,47, 663–672
16.Armando Pombeiro Maximilian N Kopylovich, European Journal of Organic Chemistry 2017,(19),
DOI:10.1002/ejoc.201601642

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23
Sulfate promoted zirconia-based mixed oxide as solid acid catalysts for
organic transformations
Sushil V. Shelkea, Sambhaji T. Dhumalb, Abhay L. Ghodkea, Meghshyam K. Patila,*
a
Department of Chemistry, Dr. Babasaheb Ambedkar Marathwada University, Sub-Campus, Dharashiv- 413501
b
Department of Chemistry, Ramkrishna Paramhansa Mahavidyalaya, Dharashiv-413501
E-mail: [email protected]
Abstract:
After gaining attention by sulfated zirconia as solid acid catalyst, various modified versions of
zirconia based catalysts have attracted attention due some extra properties associated with
stability of the catalyst. Among these modified versions, various sulfate promoted zirconia-
based mixed oxides have been reported in the literature, which includes SO42-/ZrO2-CeO2,
SO42-/SnO2-ZrO2, SO42-/TiO2-ZrO2, sulfated zirconia catalyst supported on mesostructured -
Al2O3, sulphated mesoporous La2O3–ZrO2 composite oxides, and many more. In this article,
we have summarized some of the organic transformations catalyzed by sulfate promoted
zirconia-based mixed oxides. These organic transformations includes Friedel-Crafts
benzoylation of anisole with benzoyl chloride; dealkylation of 1,3,5-tri-tertbutyl-benzene;
synthesis of coumarins via Pechmann reaction; Mannich reaction between aldehyde, ketone
and amines; synthesis of quinoxalines and synthesis of 2-aryl benzimidazoles and so on.

Keywords: Sulfate impregnation, sulfated zirconia, mixed oxide, organic transformations.

Introduction
Generally, liquid acid catalysts or homogeneous acid catalysts have been used to
perform acid catalyzed reactions. Acid catalyzed reactions are efficiently carried out by these
catalysts as these are soluble in the reaction medium and the reactant can easily reach to the
active site of the catalyst and vice-versa. Whereas, some of these catalysts are very reactive
and will not produce the product selectively. Further, the loopholes associated with the use of
these systems are non-recoverable catalysts, tedious work-up procedures, and consequently
responsible for pollution [1-5]. The scientific community was searching for a heterogeneous
counterpart of these catalysts, that can work efficiently as a catalyst, gives high yield,
recyclable, with easy and eco-friendly work-up procedure. In this regard, several
heterogeneous catalysts such as zeolite, amberlite, nafion-H, molecular sieves, sulfated
zirconia, and so on have been used as catalysts. Among these catalysts, sulfated zirconia is one
of the important catalysts. It has been used extensively as a catalyst for processes/reactions
such as alkylation, isomerization, cracking reactions, and several organic transformations [1-
6]. In addition to this, many modified versions of sulfated zirconia can be used as catalysts.
These modifications include different preparation procedures for sulfated zirconia catalysts,
which can alter the properties of the catalyst. By using templated mesoporous, sulfated zirconia
catalysts have been prepared. Other modification includes the use of MoOx or WOx as a
promoter instead of SO42 [7, 8]. Another important modification includes, sulfate
impregnation of a mixed oxide of ZrO2 with other metal oxide. In this article, we have discussed
some of the organic transformation reactions, which are catalyzed by these sulfated zirconia-
based mixed oxides.
Preparation of sulfated zirconia-based mixed oxides:
Several methods have been adopted in the literature for the preparation of sulfate
impregnated zirconia based mixed oxides. These methods includes co-precipitation, sol-gel,

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hydrothermal and so on. Reddy et al. prepared sulfated ceria-zirconia catalyst by co-
precipitation followed by sulfated impregnation [9]. Ben Hamouda and Ghorbel used sol-gel
method for preparation of sulfated zirconia supported by alumina [10]. Chakraborty et al.
employed hydrothermal method for the preparation of silica sulphated zirconia [11]. These are
some notable examples of use of different methods for synthesis of zirconia-based catalysts.
Organic Transformations by Sulfate promoted zirconia-based mixed oxide:
In this section, we have discussed some of the reactions reported by using sulfate promoted
zirconia-based mixed oxide as a catalyst.
1. Friedel-Crafts benzoylation of anisole with benzoyl chloride and dealkylation of
1,3,5-tri-tertbutyl-benzene:
Zhao et al. have been prepared sulfated zirconia supported on mesostructured -Al2O3
as a catalyst used for Friedel-Crafts benzoylation of anisole with benzoyl chloride and
dealkylation of 1,3,5-tri-tertbutyl-benzene [12].
2. Pechmann reaction and Mannich reaction:
Reddy et al. synthesized sulfate promoted CeO2- ZrO2 catalyst and utilized it for the
synthesis of coumarins via Pechmann reaction as well as Mannich reaction between
aldehyde, ketone and amines [9, 13].
3. Acetylation of alcohols and amines:
Sulfate promoted Al2O3-ZrO2 catalysts have been reported for acetylation of alcohol
and amines with acetic anhydride (Scheme 1) [14].

Scheme 1. Sulfate ZrO2-Al2O3 catalyzed acylation of alcohols and amines.

4. Synthesis of quinoxalines
Recently, Shelke et al. reported the synthesis of quinoxalines by reaction between
phenacyl bromides/benzil and o-pheneylenediamine using sulfate promoted ZrO2-TiO2
as a catalyst (Scheme 2) [15].

Scheme 2. SO42/ZrO2-TiO2 catalyzed synthesis of quinoxalines.

5. Synthesis of 2-aryl benzimidazoles
Furthermore, Shelke et al. have also been reported the synthesis of 2-aryl
benzimidazoles by reaction of o-pheneylenediamines and different aromatic aldehydes
(Scheme 3). This reaction is catalyzed by sulfate promoted ZrO2-TiO2 [16].

Scheme 3. SO42/ZrO2-TiO2 catalyzed synthesis of 2-aryl benzimidazoles.

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Conclusion:
Sulfate promoted zirconia-based mixed oxides is a one of the promising group of
catalysts in a family of solid acid catalysts. These catalysts have been used for variety of
organic transformations. These catalysts are recyclable, easy work up procedures, promotes
high yield, therefore it is an eco-friendly way to carried out many organic transformations such
as Friedel-Crafts benzoylation of anisole with benzoyl chloride; dealkylation of 1,3,5-tri-
tertbutyl-benzene; synthesis of coumarins via Pechmann reaction; Mannich reaction between
aldehyde, ketone and amines; synthesis of quinoxalines and synthesis of 2-aryl benzimidazoles.
Furthermore, these catalysts can be explored for various other organic transformations.
References:
[1] M. K. Patil, A. N. Prasad, B. M. Reddy, Curr. Org. Chem., 2011, 15, 3961.
[2] G. D. Yadav, J. J. Nair, Microporous and Mesoporous Materials, 1999, 33, 1.
[3] A. Feller. J. A. Lercher .Adv. Catal., 2004, 48, 229.
[4] A. Corma, Catal. Rev.-Sci. Eng., 2004, 46, 369.
[5] K. Arata, Green Chem., 2009, 11, 1719.
[6] G. X. Yan, A. Wang, I. E. Wachs, J. Baltrusaitis, Appl. Catal. A: Gen., 2019, 572,
210.
[7] B. M. Reddy, M. K. Patil, B. T. Reddy, Catal. Lett., 2008, 125, 97.
[8] B. M. Reddy, M. K. Patil, G. K. Reddy, B. T. Reddy, K. N. Rao, Appl. Catal. A: Gen.
2007, 332, 183.
[9] B. M. Reddy, M. K. Patil, P. Lakshmanan, J. Mol. Catal. A: Chem. 2006, 256, 290.
[10] L. Ben Hamouda, A. Ghorbel, J. Sol-Gel Sci. Tech. 2003, 26, 831.
[11] B. Chakraborty, P. Dutta, S. K. Nandi, Mater. Sci. Indian J. 2013, 9, 144.
[12] J. Zhao, Y. Yue, W. Hua, H. He, Z. Gao, Appl. Catal. A, 2008, 336, 133.
[13] B. M. Reddy, P. M. Sreekanth, P. Lakshmanan, A. Khan, J. Mol. Catal. A: Chem.,
2006, 244, 1.
[14] B. M. Reddy, P. M. Sreekanth,Y. Yamada, T. Kobayashi, J. Mol. Catal. A: Chem.,
2005, 227, 81.
[15] S. V. Shelke, S. T. Dhumal, A. Y. Karale, T. R. Deshmukh, M. K. Patil, Syn.
Commun. 2022, 52, 597.
[16] S. V. Shelke, S. T. Dhumal, T. R. Deshmukh, M. K. Patil, Lett. Org. Chem. 2023,
206, 541.

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24

Structure, Physical and Chemical Properties of Thiourea with Their

Derivatives

A.P. Mitake, S.P. Rathod,

Department Of Chemistry G. S. Gawande Mahavidyalay Umarkhed Dist. Yavatmal
E-mail : [email protected]

Abstract:

In this study six unsymmetrical thiourea derivatives, 1-isobutyl-3-cyclohexylthiourea (1), 1-tert-butyl-

3-cyclohexylthiourea (2), 1-(3-chlorophenyl)-3-cyclohexylthiourea (3), 1-(1,1-dibutyl)-3-
phenylthiourea (4), 1-(2-chlorophenyl)-3-phenylthiourea (5) and 1-(4-chlorophenyl)-3-phenylthiourea
(6) were obtained in the laboratory under aerobic conditions. Compounds 3 and 4 are crystalline and
their structure was determined for their single crystal. Compounds 3 is monoclinic system with space
group P21/n while compound 4 is trigonal, space group R3:H. Compounds (1–6) were tested for their
anti-cholinesterase activity against acetylcholinesterase and butyrylcholinesterase (hereafter
abbreviated as, AChE and BChE, respectively). Potentials (all compounds) as sensing probes for
determination of deadly toxic metal (mercury) using spectrofluorimetric technique were also
investigated.

Keywords: thiourea derivatives, enzyme inhibition,

Introduction:

It plays an important role in the construction of hetero-cycles, the compounds formed by the
insertion of one or more, similar or different hetero-atoms (other than carbon or hydrogen
atoms) in different cyclic systems. It appears as white crystals which are combustible and in
contact with fire give off irritating or toxic fumes. It acts as a precursor to sulphide to produce
metal sulphides like mercury sulphide. Thiourea is a reagent in organic synthesis, a special
branch of chemical synthesis, and is concerned with the construction of organic compounds
via organic reactions. "Thioureas" can refer to a broad class of compounds with the general
structure (R1R2N) (R3R4N) C=S. Thiourea is also called by names such as “Thiocarbamide,
and Pseudo thiourea” Thioureas belong to thioamides, a functional group with the general
structure R–CS–NR′R″, where R, R′, and R″ are organic groups. Examples include RC(S)NR2,
where R is methyl, ethyl, etc.

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Structure of Thiourea molecule

Structure of Thiourea

Properties of Thiourea

 It is an organic compound containing sulphur atoms.

 Its molar mass is 76.12 g.mol-1.
 It is a white-coloured compound.
 It exists in a solid state at room temperature.
 The melting point of thiourea is 176 – 178oC.
 Its boiling point is 150 – 160oC
 Its density is 1.405 g.ml-1.
 It is highly soluble in water. For instance, 142 g of thiourea can be dissolved in one
litre of water at 25o.
 It is slightly acidic in nature
 Its crystals are highly combustible in contact with fire.
 It has pH value of more than 3.
 It is an odourless (with no smell) compound.
 Its surface tension is 1.04 10-2 N/m.
 Thiourea on heating above 130o , forms ammonium thiocyanate. Upon cooling it
again converts into thiourea.
 Reduction – Peroxides get reduced into their corresponding Diols (chemical
compounds with two hydroxyl groups) by thiourea. During this reduction reaction, a
by-product formed which is called endo-peroxide. Endo-peroxide is a highly unstable
compound.
 Due to its non - volatile nature, it is also used in the ozonolysis of cyclic alkenes to
give carbonyl compounds.
 It reacts with alkyl halides and forms thiols.

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Uses of Thiourea

Total global production of thiourea approximately 40% is produced in Germany and another
40% in China. About 10,000 tonnes of thiourea is being manufactured globally in a year. Its
bulk production clearly proves its significance in the market. Few of its applications are listed
below:

 Thiourea Dioxide (prepared by oxidation of Thiourea with Hydrogen peroxide) is

used as a reducing agent in many chemical reactions used in textile processing.
 It is used in the production of flame-retardant resins, which prevents or slows down
the spread of fires.
 It is used as a source of sulphide, a compound of sulphur with another element. It
reacts with alkyl halides and changes them into thiols, a sulphur analogue of alcohols
that is simple it is an organic compound consisting of compounds with a sulphur
atom. For example, ethane – 1,2 – dithiol is prepared by 1,2 – dibromoethane. The
reaction is given below: C2H4Br2 + 2SC(NH2)2 → [C2H4(SC(NH2)2)2]
Br2

[C2H4(SC(NH2)2)2] Br2 + 2KOH → C2H4(SH)2 + 2OC(NH2)2 + 2KBr

 Thiourea can be used as a source of sulphide in reactions with metal ions as well. For
example, mercury sulphide is formed when mercury ion reacts with thiourea in
presence of water and heat. The reaction is given below:

Hg2+ + SC(NH2)2 + H2O → HgS + OC(NH2)2 + 2H+

 Thiourea condenses with - dicarbonyl and forms pyrimidine derivatives. It is used in

vulcanization accelerators.
 It is used as an auxiliary agent.
 It is used in silver – gelatine photographic prints, diazo paper, light-sensitive
photocopy paper, etc.
 It is used in many electroplating processes such as Clifton – Philips and Beaver bright
electroplating, etc.
 For copper printed circuits, tin (II) chloride solution is used. Thiourea is also used in
the solution.
 It is used in silver cleaning products such as Tarn x which contains thiourea, sulfamic
acid and detergent.
 It is used in gold and silver leaching. Lixiviants are used for this purpose and thiourea
is an important ingredient of these. Lixiviant is a liquid medium used in
hydrometallurgy to selectively extract the desired metal from the ore or mineral.

 Even short exposure to the Thiourea causes severe irritation and temporary skin
infection.
 125mg/kg of thiourea can be deadly for rats on oral ingestion.
 On chronic exposure, thiourea can cause a goitrogenic effect on humans. Goitrogens
can reduce the thyroid's ability to produce the hormones your body needs to function
normally.
 Enlargement of the thyroid gland is known as goiter disease. In this disease,
goitrogens disrupt the production of thyroid hormones. This stimulates the pituitary
gland to release TSH and TSH promotes the growth of thyroid tissues.

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Six Different Derivatives of Thiourea

Figure 1: Structure of unsymmetrical thiourea derivatives, 1-cyclohexyl-3-(iso-butyl)thiourea

(1), 1-cyclohexyl-3-(tert-butyl)thiourea (2), 1-cyclohexyl-3-(3-chlorophenyl)thiourea (3), 1-
phenyl-3-(1,1-dibutyl)thiourea (4), 1-phenyl-3-(2-chlorophenyl)thiourea (5) and 1-phenyl-3-
(4-chlorophenyl)thiourea (6) used in this study.

Material and Method:

All reactions were carried out under aerobic atmosphere and no special precautions were taken
to exclude air or moisture during experiments. All chemicals were purchased from Vinit
Chemicals, Nanded and were used without further purification. Thiourea derivatives described
in this study are also commercially available in the marked, we reproduced them in the
laboratory, they were obtained as crystalline material. The formation of compounds was
confirmed with the help of available spectroscopic techniques, such as ATR-FTIR
spectrometer and 1H- and 13C NMR spectra (in order to assist readers these data is provided
below). Thiourea derivatives were prepared by reactions as previously reported [1,2].

Experimental Work
1.1. 1-cyclohexyl-3-isobutylthiourea, 1
Compound 1was synthesized by dropwise addition of cyclohexyl isothiocyanate (1 mL, 7.05
mmol) to isobutyl amine (0.7 mL, 7.05 mmol) in analytical grade acetone. The reaction mixture
was allowed to stir over night at room temperature. The reaction led to the formation of
precipitates and the progress was monitored by TLC. Solid material was separated from the
mother liquor, washed with petroleum ether for removal of unreacted compounds and possible
impurities. The solid was re-dissolves in ethanol and kept for slow evaporation. Spectroscopic
datawere collected and structure of the desired compound was deduced.
Yield = 70 %; Molecular formula = C11H22N2S; m.p. = 102-105˚C; FT-IR(ATR): ν(cm−1)
= 3431br (NH), 3214br (N-H), 2930s, 2850w (CH), 1617 (C=S), 1379 (NCN), 1448 (CSasy);
1
H-NMR (300 MHz, DMSO-d6) δ(ppm)= 0.85 (d, J(1H,1H) = 6.7 Hz, 6H, Me), 1.16,
1.25, 1.53, 1.63, 1.82 (overlapping multiplets of aliphatic protons), 3.18 (br, 1H, NH),
3.94(br,1H, NH). 13C-NMR (75 MHz, DMSO-d6) δ(ppm) = 20.1 (2C), 24.5 (2C), 25.1 (1C),
27.7 (1C), 32.3 (2C), 50.9 (1C), 51.6 (1C), 181.3 (C=S).

1.2. 1-cyclohexyl-3-(tert-butyl) thiourea, 2

In the similar manner as described above compound 2was prepared by mixing tertbutylamine
(1 mL, 9.5 mmol) with a solution of cyclohexyl isothiocyanate (1.35 mL, 9.5

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mmol) in dry acetone. Compound 2 was obtained as a colorless solid in ethanol.

Yield = 68 %; Molecular formula = C11H22N2S; m.p.= 138-140˚C; FT-IR (ATR): ν(cm−1)
= 3421br, 3367br (N-H), 3050w, 2927w (C-H), 1636s (C=S), 1385 (NCN), 1544 (CSasy);
1HNMR (300 MHz DMSOd6) δ(ppm) = 1.03 (m, 2H, CH2), 1.20 (m, 4H, CH2), 1.61 (m,
4H,
CH2), 1.41 (s, 9H, Me), 1.83 (m, 1H, CH), 2.50 (p, 1H, CH), 3.16 (d, 1H, NH), 4.08 (br, 1H,
NH); 13C-NMR (75 MHz, DMSO-d6) δ(ppm) = 24.4, 25.2, 29.0, 32.3, 50.6, 52.0, 180.2
(C=S).

1.3. 1-cyclohexyl-3-(3-chlorophenyl) thiourea, 3

A known amount of 3-chloroaniline (0.5 mL, 4.7 mmol) in 15 mL analytical grade acetone
and equimolar amount of cyclohexyl isothiocyanate (0.6 mL, 4.7 mmol) was reacted together
following the same method. Colourless crystals of 3were obtained in EtOH in few days, were
separated from the mother liquor, FT-IR and NMR data were collected and suitable crystal of
the compound was mounted for X-ray data collection and structure confirmation.
Yield = 66 %; Molecular formula = C13H17ClN2S, m.p. = 115-120˚C, FT-IR(ATR):
ν(cm−1) 3295br, 3206br (N-H), 3060w (Ar, C-H), 2993w, 2849w (C-H), 1700s (C=S), 1534s
(NCN), 1435s (CSasy); 1H-NMR (300 MHz, DMSO-d6) δ(ppm) = 1.26 (m, 4H, CH2), 1.67
(m, 4H, CH2), 1.92 (m, 2H, CH2), 1.99 (m, 1H, CH), 4.17 (br, 1H, NH), 7.10, 7.26, 7.31,
7.82 (m, m, m, m, 4H, CH, Ph),9.45 (br, 1H, NH); 13C-NMR (DMSO-d6) δ(ppm) = 25.0,
31.9, 39.8, 52.5, 121.0, 122.1, 123.5, 130.0, 132.9, 141.8, 179.5 (C=S).

1.4. 1-phenyl-3-(1,1-dibutyl) thiourea, 4

Phenyl isothiocyanate(1mL,8.3mmol) and dibutyl amine (0.88 mL, 8.3 mmol) in 20 mL

acetone was treated as discussed above. Colourless crystals of the desired compound 4
were obtained in the same solvent at ambient temperature.
Yield = 95%; Molecular formula = C15H24N2S, m.p. = 85-87˚C; FT-IR (ATR 400-4000
cm−1) ν (cm−1) = 3700br (N-H), 2335w (C-H), 1529s (C=C), 1452s (NCN), 1205s (CSasy);
1HNMR (300 MHz, CDCl3), δ (ppm) = 0.96 (t, 6H, Me), 1.37 (m, 4H, CH2), 1.69 (pent, 4H,
CH2), 3.65 (t, 4H, CH2N), 7.05(br, 1H, NH), 7.18, 7.31 (m, m, 5H, Ph); 13C-NMR (75
MHz, CDCl3) δ(ppm) = 13.7, 20.1, 29.4,51.4, 125.5, 125.6, 128.5, 139.8, 181.0 (C=S).

1.5. 1-Phenyl-3-(2-chlorophenyl) thiourea, 5

In the similar manner as described above by reacting 2-chloroanaline (0.87 mL, 8.3
mmol) with phenyl isothiocyanate (1 mL, 8.3 mmol), the product 5 was obtained as
colourless crystalline solid by recrystallization from ethanol. In the whole reaction process,
the
reaction was monitored by TLC until single spot product was formed.
Yield = 69 %; Molecular formula = C13H11ClN2S; m.p. = 90-92˚C; FT-IR (ATR) :ν(cm−1)
= 3700br (NH), 2335w (C-H), 1528s (C=S), 1452s (NCN), 1442s (CSasy); 1H-NMR
(DMSOd6) δ(ppm) = 7.16, 7.23-7.29, 7.32-7.39, 7.53, 7.63 (m, m, m, m, m, 9H, Aromatic
protons), 9.43 (br, 1H, NH), 10.02 (br, 1H, NH); 13C-NMR (DMSO-d6) δ(ppm) = 123.8,
124.7, 127.2, 127.5, 128.6, 129.4, 129.8, 130.0, 136.4, 139.2, 180.3 (C=S).

1.6. 1-phenyl-3-(4-chlorophenyl) thiourea, 6

The derivative 6, was obtained as colourless solid by treating 4-chloroaniline (0.8 mL,
8.3 mmol) and phenyl isothiocyanate (1.0 mL, 8.3 mmol) in the same manner as reported

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in literature [3]. Crystals were obtained in ethanol at room temperature by slow evaporation
method.
Yield = 70 %; Molecular formula = C13H11ClN2S; m. p. = 95-99˚C; FT-IR (ATR): ν(cm−1)
= 3436-3100br (N-H), 3000w (C-H), 1637s (C=S), 1549s (NCN), 1490s (CSasy); 1HNMR
(DMSO-d6) δ(ppm) = 7.14, 7.37, 7.52 (m, m, m, 10H, Aromatic protons, NH), 9.88 (br, 1H,
NH); 13C-NMR (DMSO-d6) δ(ppm) = 123.8, 124.6, 125.3, 128.3, 128.5, 138.5, 139.3, 179.3
(C=S).

3. NMR spectra of compounds 1–6

The 1H- and13C-NMR of all compounds were recorded on Bruker AM 300 MHz spectrometer.
In the 1H-NMRspectra, all the aliphatic and aromatic protons appeared in their characteristic
and expected region, 0.8-3.4 ppm and 7.0-7.9 ppm, respectively [4]. The NMR data analysis
of all compounds reveal that the reaction is straightforward and no side reactions were
observed. As each compound contains two types of N-H protons with different environment
(except 4), and accordingly two signals were observed for each of them. The N-H proton close
to aromatic ring was observed in up-field region while the other appeared in down-field region.
From 13
C-NMR spectra of compounds 1-6, it is evidenced that all the aliphatic and aromatic carbons
appeared below 50 ppm and 120-140 ppm, respectively. The chemical shift corresponding to
C=S bond was noticed in the region of ≈180 ppm, the presence of this carbon in the spectra of
compounds 1-6 provides enough justification regarding the formation and presence of thiourea
derivatives. Other carbon atoms belong to molecules of respective compounds appeared in their
characteristic regions.

Table S1. UV-Visible spectroscopic data of compounds 1–3, 5and 6.

Sr.No Compound Concentration (µg mL−1) Scan range λmax(nm)
(nm)
1 Comp -1 200-400 296

2 Comp - 2 200-400 294

3 Comp – 3 2 200-400 298

4 Comp -5 200-400 294

5 Comp - 6 200-400 295

Result and Conclusion:

Compounds 1–6 are structurally very simple; they are accessible in the market and can also be
obtained in laboratory under ambient conditions [5]. Compounds structurally analogous to 1–
6 are good corrosion inhibitors in acidic medium [6] and show excited results in affording metal
sulfides for useful applications [7]. Compound 5 has been used as starting precursors for the
preparation of benzothiazole derivatives under catalyst free conditions [8]. There are several
other reports wherein simple thiourea molecules after certain modifications have shown
efficiency as bioactive compounds [9]. Compound 6 has already been used for its inhibitory
activity against melanin B16 cells and mushroom tyrosinase and its synthesis has been
reviewed [10,11]. The IC50 value as melanin B16 inhibitor was promising, 3.4 μM, while it
exhibited moderate potency as mushroom tyrosinase inhibitor. The enzyme inhibition (AChE

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and BChE) and metal sensing capability of compounds 1–6 (Figure 1) have not been reported
so far. Since they are already in the field of bioorganic chemistry therefore these studies will
help exploring their multidimensionality applications within a single system or organism
whatever the case may be.

References:

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25

Study of Proton and Metal-Ligand Stability Constants of hydroxy

Substituted Chalcone Complexes pH-Metrically

Mohd. Wajid Shaikh Mohd Waris

Department of Chemistry, Yashwantrao Chavan Arts and Science Mahavidyalaya
Mangrulpir, Dist. Washim 444403, Maharashtra, India.
[email protected]

ABSTRACT

In the present work, we have investigated the proton-ligand stability constants and
metal-ligand stability constants of ligand 1-(5-bromo-2-hydroxyphenyl)-5-phenylpenta-2,4-
dien-1-one (L3) with transition metal ions like Cr (III) and Nd (III), which were determined
pH-metrically at 0.1 M ionic strength. (30±10C) in a 70% dioxane-water mixture by the
Bjerrum method as adopted by Calvin Wilson. 1:1 and 1:2 complexes were formed between 1-
(5-bromo-2-hydroxyphenyl)-5-phenylpenta-2, 4-dien-1-one (L3), Cr (III), and Nd (III). The
values of pK and log k were evaluated and compared with the resultant data.
Keywords: Substituted chalcone, Dioxane – water mixture, stability constant.
1. Introduction
The stability constant, also known as the formation constant or association constant, is
a measure of the strength of the bond formed between a ligand and a metal ion in a complex.
It quantifies the stability or tendency of the complex to form and remain intact in a solution.
Stability constants are determined through various experimental techniques, such as
spectrophotometry or potentiometry. These constants are expressed as equilibrium constants
and are typically denoted by the symbol K. The higher the stability constant, the more stable
the complex is. Stability constants are important in various fields of chemistry, including
coordination chemistry, biochemistry, environmental chemistry, and analytical chemistry.
They play a crucial role in understanding the behavior of metal-ligand complexes in solution,
their formation, and their potential applications.
The value of a stability constant depends on several factors, including the nature of the
metal ion and ligand, the pH of the solution, temperature, and ionic strength. These constants
provide valuable insights into the thermodynamics of complex formation and can be used to
predict the stability of a complex under different conditions. Overall, stability constants are a
fundamental concept in chemistry that helps to understand the formation and stability of metal-
ligand complexes, and their importance extends to various areas of scientific research and
applications.
A significant contribution was made to the field of stability constants of organic ligan
ds and their metal complexes by Bjerrum1 and Calvin2. The Schiff base N-[2-hydroxy-1-
napthalydene]-2-methoxyanilline (3) was formed through condensation of 2-hydroxy-1-
napthalydene with 2-methoxyanilline. It was studied for its formation of transition elements
and its stability constants, with the order of stability constant being Cu > Ni > Zn > Mn > CO3.
Jagtap's study examined the stability constant of Ni (II) with substituted pyrazole carboxylic
acid derivatives at 298K in a 70% DMF-water mixture. 4 The stability constants of transition
metals, including Cu (II) and Co (III) complexes with Schiff bases, were evaluated for anti-
diabetic drugs. They were determined using the Calvin Bjerrum titration technique and

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discussed in terms of order, ligand basis, and correlations. 5 Thakur et al. 6 studied the stability
constant of Schiff bases with lanthanum using pH metric titration and the Calvin-Bjerrum
method. They calculated thermodynamic parameters like Gibb's free energy change, entropy
change, and enthalpy change. Thorat et al. 7 studied the complex formation between Pr (III)
and Sm (III) metal ions and 3-(2-hydroxy-3-nitro-5-methylphenyl)-5-(3-nitrophenyl)
isoxazoline, 4-chlorophenyl isoxazoline, and 2-furyl isoxazoline. The study investigated the
interaction of metal ions with the organic ligand N-[(E)-(4-Hydroxy-3-methoxyphenyl)
methylene] isonicotinohydrazide. The ligand was synthesized using an anti-mycobacterial
agent and aromatic aldehydes. The formation of these ligands was confirmed through various
tests, with the stability constants arranged in a specific order. 8 The stability constants and
thermodynamic parameters of transition metal complexes of substituted aminothiazole Schiff
bases have been studied by R.P. Giram et al.9
2. Experimental methodology
2.1Material and Methods
All chemicals used are AR-grade. The ligand (L3) was synthesized in the laboratory by
the reported protocol10. The stock solution of the ligand was prepared by dissolving the required
amount of ligand in a 70% (dioxane and water) mixture.
2.2 General procedure

Types of Titrations

i) Free acid HNO3 (0.01 M)

ii) Free acid HNO3 (0.01 M) and ligand (20 x 10-4M)

iii) Free acid HNO3 (0.01 M) and ligand (20 x 10-4) and metal ions (4 x 10-4M) against a
standard 0.1N NaOH solution. The ionic strength of all the solutions was maintained at 1M
by adding the appropriate amount of KNO3 solution. All the titrations were carried out in a
70% (dioxane and water) mixture, and the readings were recorded for each 0.1 ml addition.
The graph of the volume of alkali added (NaOH) against pH was plotted. The ligand involved
in the present work may be considered a monobasic acid having only one dissociable H+ ion
from the phenolic OH group, and it can therefore be represented as HL. The dissociating
equilibrium can be shown as.

HL⇌H+ + L-
By the law of mass action, we have,
K = [HL] / ([H+] [L-]) (1)
Where, the quantities in bracket denote the activities of the Species at equilibrium.
3. Result and Discussion
3.1 Calculation of Proton-Ligand Stability Constant (nA)
The plots between the volume of NaOH and the pH of the solution were used to
determine the proton ligand stability constant (representing the replacement of H+ ions from
the functional group of the ligand with respect to the pH value). The horizontal difference (V2-
V1) was measured accurately between the titration curves of free acid and acid + ligand. It was

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used to calculate the formation number n at various pH values and a fixed ionic strength μ =
0.1 M using Irving and Rossotti’s equation [1, 2].

_ (E0+N) (V2-V1)
nA = γ -------------------- (2)
(V0 + V1) TL0

Where V0 is the initial volume of the solution. E0 and TL0 are the initial concentrations
of the mineral acid and ligand, respectively. V1 and V2 are the volumes of alkali of normality
N during the acid and ligand titration at a given pH. γ is the replaceable proton in the ligand.
The data on nA obtained at various pHs, along with the horizontal difference for some
representative systems, are represented in Table 1. The metal ligand ligand formation number
(n) is estimated by Irving-Rossotti's equation.
_ (E0+N) (V3-V2)
n= -------------------- (3)
(V0 + V2) Tm0

Where the notations have the same meaning as given in the earlier equation. The
horizontal difference (V3-V2) between the metal complex (A+M+L) and reagent (A+L) curve
is used to evaluate the value of n using Irving Rossotti’s equation.

Table 1 Proton ligand stability constant (pK)

Ligand System pK
Half integral Point wise
method method
L3 1-(5-bromo-2-hydroxyphenyl)-5- 7.7871 6.4506
phenylpenta-2,4-dien -1- One

Table 2: Metal-ligand stability constant (log K)

System LogK1 LogK2 ∆ LogK
Cr (III)+L3 5.7419 3.3126 2.4293
Nd (III)+L3 5.5310 3.5278 2.0032

Conclusion
From the titration curves, it is observed that the departure between the acid-ligand
(A+L) curve and the acid-ligand-metal (A+L+M) curve for all systems started at pH 3.6. This
indicated the commencement of complex formation. Also, the change in color from yellow to

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orange in the pH range from 3.9 to 8.6 during titration showed the complex formation
between metal and ligand.

The difference between LogK1 and LogK2 is less than 2.5, indicating the simultaneous
formation of 1:1 and 1:2 complexes. When the difference is greater than 2.5, a stepwise
complex formation takes place. 11. From Table 2, it is observed that in the all systems Cr (III)-
L3 and Nd (III) + L3, the difference between log K1 and log K2 is sufficiently small, which
indicates the simultaneous formation of complexes between metal ions and ligands takes place.

Acknowledgement
The author is also thankful to the Dr. R.D. Deshmukh, Principal, R.D.I.K. & N.K.D.
College, Badnera, for providing useful guidance and facilities throughout the research work.
Reference
1 Bejerrum J. (1941). Metal Ammine formation in aqueous solution, haase, copengegan. 296p, 8(9), 20
(1982).

2 Calvin M, Wilson KW. (2003). stability of chelate compounds, J Am. Chem. Soc, 1945, 67.

3 Jadhav S, Rai M, Pardeshi R.K, Farooqui M. (2015). Determination of stability constant of metal ligand
equilibria with special reference to Schiff base and transition elements. Der Pharmacia Lettre, 7 (12):
316-320. (http://scholarsresearchlibrary.com/archive.html)

4 Jagtap V. S (2019). Determination of Stability Constant of Some Coordination Compound by pH

Metric Technique. Pramana Research Journal, 9 (8): 22-29. https://pramanaresearch.org/

5 Parihar R. T, Quazi S. A. (2019). Study of proton & Metal ligand stability constant of Schiff base
complexes in 70% DMF solvent media pH metrically. Journal of Emerging Technologies and
Innovative Research, 6 (1): 136-144.

6 Thakur S.V, Farooqui M.N, Jadhav R. L, Joshi S. U. (2021). Complexation of La (III) metal ion with
novel Schiff bases. Thermodynamic study. Journal of Advanced Scientific Research, 12(2): 133-136.
http://www.sciensage.info

7 Thorat, S.A, Thakur. S.D. (2015). Metal ligand stability constant of substituted 3,5-Diaryl isoxazolines
complexes in 70% dioxane solvent media. J. Curr. Chem. Pharm. Science, 5(2): 67-74.

8 Omar Z. T, Jadhav S. L, Kayande D. D, Rai M. J. (2019). pH-metric Study of Metal-ligand Stability

Constant of Transition Metal Complexes with Pharmacologically Active Ligand N-[(E)-(4-Hydroxy-3-
methoxyphenyl) methylene] isonicotinohydrazide. Curr. Pharm. Res, 407, 58-65.
https://www.researchgate.net/publication/333365049

9 Giram, R.P, Muthal, B.N, More, P.G. (2019). The study of stability constants and Thermodynamic
parameters of transition metal complexes of substituted aminothiazole Schiff Bases. The Pharma
Innvation Journal, 8(1): 249-252. https://dx.doi.org/10.22271/tpi

10 Swapnil, K.W, Kakade, K.P. (2013). Synthesis and characterization of substituted 5-bromo-2-
benzylidine-1-benzofuran-3-one and its structural determination. International Journal of chemical and
pharmaceutical science, 4(3): 119-122.

11 Deolankar, D. S, Deshpande, Y.H. (1987). La(III), Pr(III), Gd(III), Tp(III) and Dy(III)

Complexes of aryl furyl and aryl thienyl β-diketones. Indian Journal of Chemistry, 26(A): 68-69.
http://nopr.niscpr.res.in/handle/123456789/47785

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26
Ecofriendly synthesis and characterization of Ethyl 2-imino-6-methyl-4-
substituted phenyl-1, 2, 3, 4-tetrahydropyrimidine-5-carboxylate
compounds
Nilesh B. Jadhav*
Department of Chemistry, Jagadamba Mahavidyalaya, Achalpur city
Email Id: [email protected]

Abstract: Ethyl acetoacetate, substituted aromatic aldehyde, and guanidine were the three
components of a one-pot, three-component reaction that produced ethyl 2-imino-6-methyl-4-
substituted phenyl-1, 2, 3, 4-tetrahydropyrimidine-5-carboxylate compounds [4a-c]. The
reaction is effectively supported to provide the intended products, typically in high yields and
within a brief timeframe. The newly synthesized chemicals structures were clarified by
elemental and spectrum investigations. Using TLC, the purity of each compound was
ascertained.
Keywords: Pyrimidine, Guanidine.
1. Introduction:
Heterocycles that include nitrogen are of synthetic importance since they belong to a significant
class of both natural and artificial compounds, many of which have shown beneficial biological
activity. the fascination with six-membered structures with two nitrogen atoms at positions
one and three. In heterocyclic synthetic chemistry, the One Pot Multicomponent Reactions
(MCRs), in which several reactions occur often in a single synthetic operation, have been
heavily employed to generate carbon-carbon bonds. Similar reactions provide a multitude of
options for efficiently building relatively complex molecules in a single phase of the process,
so avoiding the laborious purification processes and allocating savings of both reagents and
solvents. For this reason, understanding their biological conditioning and therapeutic
applications is crucial for scientists working on drug discovery processes3. We have conducted
the reactions in water, which is notable for its use as a non-flammable, non-hazardous, and
green medium, based on the principles of green chemistry.
Pyrimidines have biological significance that motivates researchers to work on them. These
activities include analgesic1,2,9, antimicrobial activity3,4,7 , anticonvulsant5, antioxidant
activity5, antimycobacterial6, antitumor7, antiviral8, anticancer9,10 and antifolate10.
2. Material and methods:
An uncorrected melting point was determined using open capillary tubes. Using an FTIR
Agilent Technologies spectrometer at 4000-650, the FTIR spectra were recorded. Using
tetramethyl silane (TMS) as an internal reference and d6-DMSO/CDCl3 as the solvent, the 1H
NMR spectra were recorded on a Bruker spectrophotometer operating at 500 MHz. The units
of chemical changes were ppm. We bought all of the solvents and reagents from Sigma Aldrich
Chemicals Pvt Ltd. To maximize the reaction's purity and completeness, TLC was employed.

2.1 Synthesis Process in General:

2.1.1 Synthesis of phenyl-1,2,3,4-tetrahydropyrimidine-5-carboxylate by ethyl 2-imino-6-
methyl-4 substitution[4a-c]:
Substituted aromatic aldehyde (0.01mol), ethyl acetoacetate (0.01mol), and guanidine
(0.01mol) were added to 15 ml of distilled water in a dry 50 mL round-bottom flask, and the
mixture was manually agitated for two minutes. The reaction mixture was then heated for an

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hour at 90 degrees Celsius in a water bath. A solid began to deposit as the reaction went on,
and an hour later the flask was full with solid. Using a spatula, the solid was carefully removed
from the conical flask. One milliliter of cold water was used to wash the yellow solid. From
ethanol, the crude product was recrystallized to remove impurities.

Scheme-1
3. Physiochemical and analytical data for compounds:
3.1 Ethyl 4-(4-hydroxy phenyl)-2-imino-6-methyl-1, 2, 3, 4-tetra hydro pyrimidine-5-
carboxylate(4a):
M.F: C14H17N3O3. M.W:278. M.P: 124–126 oC. Yield: 77.35%. FT-IR: 1510(Ar-
C=C),1631(C=N), 1715(C=O), 2887(C-H), 3199(N-H), 3402(Ar-OH). 1H-NMR (500 MHz,
DMSO d6, δ ppm): 7.81(s, 1H, =N-H), 6.91-6.71 (Ar-H), 4.50(S, 1H, R2-NH), 4.03(q, 2H, -
CH2), 1.25(t, 3H, -CH3), 2.27 (s, 3H, Ar-CH3), 9.1 (s, 1H, Ar-OH). Anal. Calcd. (in %): C,
61.07; H, 6.20; N, 15.21. Found: C, 61.11; H, 6.281; N, 15.35.
3.2 Ethyl 4-(4-cyano phenyl) -2-imino -6-methyl-1, 2, 3, 4-tetra hydro pyrimidine -5-
carboxylate(4b):
M.F: C15H16N4O2. M.W:285. M.P: 112–114 oC. Yield: 76.81%. FT-IR: 1502(Ar-C=C),
1628(C=N), 1714(C=O), 2265(Ar-CN), 2888(C-H), 3195(N-H). 1H-NMR (500 MHz, DMSO
d6, δ ppm): 7.81(s, 1H, =N-H), 7.76-7.28 (Ar-H), 4.48(S, 1H, R2-NH), 4.02(q, 2H, -CH2),
1.20(t, 3H, -CH3), 2.19(s, 3H, Ar-CH3). Anal. Calcd. (in %): C, 63.39; H, 5.45; N, 19.83.
Found: C, 63.35; H, 5.63; N, 19.69.
3.3 Ethyl 2-imino-6-methyl -4-phenyl-1, 2, 3, 4-tetra hydro pyrimidine- 5-carboxylate(4c):
M.F: C14H17N3O2. M.W: 260. M.P: 112–114 oC. Yield: 73.80%. FT-IR: 1509 (Ar-C=C),
1628(C=N), 1717 (C=O), 2882(C-H), 3199(N-H). 1H-NMR (500 MHz, DMSO d6, δ ppm):
7.81(s, 1H, =N-H), 7.28-7.17 (Ar-H), 4.59 (s, 1H, R2-NH), 4.12 (q, 2H, -CH2), 1.20 (t, 3H, -
CH3), 2.31 (s, 3H, Ar-CH3). Anal. Calcd. (in %): C, 64.79; H, 6.59; N, 16.25. Found: C, 64.15;
H, 6.29; N, 16.89.
4. Conclusion:
The main focus of this research work was pyrimidine derivatives have been synthesized based
on green chemistry principles (multicomponent synthesis) using water as a solvent. The
structures of synthesized compounds were confirmed by FTIR, 1HNMR and elemental

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analysis. The reaction is effectively supported to provide the intended products, typically in
high yields and within a brief timeframe

5. References:
1) Huang, Zhibin, et.al. "Efficient one-pot three-component synthesis of fused pyridine
derivatives in ionic liquid." ACS Combinatorial Science 13, no. 1 (2011): 45-49.
2) Arora, Neha, et.al. "Synthesis and analgesic activity of novel pyrimidine
derivatives." Synthesis 11, no. 1 (2011): 010.
3) Saczewski, et.al. "Biological activities of guanidine compounds." Expert opinion on
therapeutic patents 19, no. 10 (2009): 1417-1448.
4) Rawat, et.al. "synthesis and antimicrobial activity of some new heterocyclic guanidine
derivatives." world journal of pharmacy and pharmaceutical sciences 5, no. 9 (2016):
1325-1337.
5) Mohana, et.al. "Synthesis and biological activity of some pyrimidine derivatives." Drug
invention today 5, no. 3 (2013): 216-222.
6) Kumar, et.al. "Syntheses of novel antimycobacterial combinatorial libraries of
structurally diverse substituted pyrimidines by three-component solid-phase
reactions." Bioorganic & medicinal chemistry letters 12, no. 4 (2002): 667-669.
7) Baraldi, et.al. "Antimicrobial and antitumor activity of N-heteroimmine-1, 2, 3-
dithiazoles and their transformation in triazolo-, imidazo-, and
pyrazolopirimidines." Bioorganic & medicinal chemistry 10, no. 2 (2002): 449-456.
8) Nasr, et.al. "Pyrido [2, 3‐d] pyrimidines and Pyrimido [5′, 4′: 5, 6] pyrido [2, 3‐d]
pyrimidines as New Antiviral Agents: Synthesis and Biological Activity." Archiv der
Pharmazie: An International Journal Pharmaceutical and Medicinal Chemistry 335, no.
6 (2002): 289-295.
9) Sondhi, et.al. "Anticancer, anti-inflammatory and analgesic activity evaluation of
heterocyclic compounds synthesized by the reaction of 4-isothiocyanato-4-methylpentan-
2-one with substituted o-phenylenediamines, o-diaminopyridine and (un) substituted
o." Australian Journal of Chemistry 54, no. 1 (2001): 69-74.
10) Gangjee, et.al. "Synthesis, antifolate, and antitumor activities of classical and
nonclassical 2-amino-4-oxo-5-substituted-pyrrolo [2, 3-d] pyrimidines." Journal of
medicinal chemistry 44, no. 12 (2001): 1993-2003.

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27

Effect of Fe Dopant on Structural & Electrical Properties of Copper Oxide

Nanoparticles

N.N.Gour , A.P.Pinjarkar , H.G.Wankhade , P.R.Padole

Shri Shivaji Science College Amravati Corresponding Author: [email protected] &
[email protected]

Abstract: Nanocrystalline Copper oxide (CuO2) was synthesized by sol-gel citrate method.
Further it was modified by doping Fe in it. Structural and Electrical study was carried out for
modified and unmodified CuO2. From X-ray diffraction (XRD) studies it was observed that
when CuO2 doped with 2% Fe its Average Crystallite Size Increases from 41.32 nm to 54.51
nm on further doping with 0.1% & 0.3% Fe Its Average Crystallite Size decreases to 34.15
nm& 31.35 nm respectively. The electrical study of the nanocrystalline Copper Oxide is
checkedfor different doping amount and also at different temperature for every sample. From
the studyit is observed that, conductivity changes with the concentration of dopant and also
varies withtemperature.
Keywords: Copper oxide, X-ray diffraction, crystallite size, Electrical Properties &
Conductivity.

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28
Preliminary Phytochemical Studies of Cesalpinia crista L.
Nitin G. Asole, Ramesh T. Parihar* and Anand S. Jadhao
Bapumiya Sirajoddin Patel Arts, Commerce and Science College, Pimpalgaon Kale, Tq. Jalgaon
(Jamod), Dist. Buldhana – Maharashtra
*Vidnyan Mahavidyalaya Malkapur, Tq. Malkapur, Dist. Buldhana – Maharashtra
Email Id: [email protected]

Abstract:
Caesalpinia crista L. belongs to Fabaceae and commonly known as ‘sagargoti’ in
Marathi. It is widely employed across the Asian and African continent for treatment of various
kinds of diseases and disorders like abdominal pain, amenorrhea, cystic fibrosis, diabetes,
leucorrhea, malaria fever, rheumatoid arthritis. The aim of present study was to investigate the
presence of phytochemicals of root, stem, leaf and fruit of Cesalpinia crista L. Ethanol,
acetone, hexane and petroleum ether extract of selected material were screen for the presence
of different secondary metabolites, it revealed the existence of alkaloid, steroids, phytosterol,
tannin, flavonoids, and glycosides. The findings of the present study provided evidence that
crude organic solvent extracts of Caesalpinia crista L. contain medicinally important bioactive
compound, it justifies their use in traditional medicines for the treatment of various kinds of
diseases.
Key words: Caesalpinia crista L., organic solvents, phytochemicals
Introduction:
Caesalpinia crista L. an important medicinal plant belongs to Fabaceae
(Caesalpiniaceous), different parts of C. crista has several traditional medicinal applications
and health benefit effects C. crista generally presents in tropical areas and found almost in
every part of India and Pakistan (Bhanderi, et al., 2022). Root, leaves, seed and bark is used
in the treatment of different kinds of diseases such as pulmonary tuberculosis, malaria,
pneumonia, colic fever, intermittent fever, swelling, menstrual complaints, skin diseases, tonic
and as a uterine stimulant, to cleanse uterus and also alleviates edema and abdominal pain
during this period. Various plant parts of C. crista traditionally used as antipyretic, periodic,
tonic and vesicant for the treatment of backache, constipation, skin diseases, gynecological
disorders, piles, ulcers and rheumatism (Suryawanshi and Patel, 2011; Patil, K. S. 2005).
The extracts different parts of C. crista have been reported to have anthelmintic
activity antimalarial activity, antioxidant activity, anti-amyloidogenic, nootropic activity and
cytotoxic activity (Sadiya Afreen et al., 2016). Literature on C. crista revealed presence of
different compounds and most of them belong to diterpenes group. Several traditional
medicinal plants have been reported to have strong antimicrobial properties and some of them
have already been used in the treatment of animals and people suffering from viral infection
(Mandal, et al., 2011).
Many plant and herbs that have potential as novel antiviral agents have been reported
and wide spectrum of active secondary metabolites including alkaloids, coumarins, flavonoids,
furyl compounds, lignans, polyphenolics, sulphides, saponins, terpenoids, thiophenes, proteins
and peptides have been identified (Srinivasa et al., 2003). The decoction of Caesalpinia crista
root has been used for its significant health benefits to treat rheumatism, backache and as a
tonic. In Ayurveda Indian Medicine System heartwood is bitter, astringent, constipating,

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hemostatic and sedative and it is useful in the treatment of burning sensation, convulsions,
diabetes wounds, diarrhoea, dysentery, epilepsy, leucorrhoea, leprosy, menorrhagia,
haemorrhages, ulcers, skin diseases, (Kirtikar and Basu,1989).
Caesalpinia crista is a large woody climber height up to 15 m grows mainly on the
river bank in tidal forests near eastern seacoast from Orissa and western sea coast from Konkan
southwards and grown throughout India up to 800 meters elevation. Branches are finely grey-
downy, armed hooked and straight hard yellow prickles on them. Bark is black, branchlets are
glossy with recurved prickles. leaflets; 2-3 pair coriaceous, acute or obtuse, ovate or elliptic,
upper surface shining, lower dull large, leaves bipinnate with recurved prickles at the base of
pinnae, 2-5 pairs, often armed. Seeds of the plant are round, dry and hot, up to 1.2 cm in length,
protected in a glossy yet very hard coat that may be green or ash gray in color, surface of the
kernel is ridged and is furrowed and is about 1.23-1.75 cm in diameter (Preedy et al., 2011;
Pooja Upadhyay et al., 2019).
Materials and Methods
Collection and Identification of materials:
The plant material i.e. roots, stem, leaves and fruits of Caesalpinia crista L., were
collected from the botanical garden of Bapumiya Sirajoddin Patel Arts, Commerce and Science
College, Pimpalgaon Kale, Tq. Jalgaon (Jamod), Dist. Buldhana – Maharashtra. The samples
were collected during the year 2022-23. The Herbarium of Caesalpinia crista L., was prepared
and authenticated from Department of Botany, Vidnyan Mahavidyalaya Malkapur. The root,
stem and leaves were washed and kept for shade drying for 7-8 days or till it get the constant
weight, whereas mature fruit was shade dried till get the constant weight. The fine powder of
selected material were prepared with the help of mortle and pistle and finely powdered with
the help of electric mixture grinder, prepared powder were stored in air tight container bottles
at room temperature until used. 5 gram of the plant material powdered was wrapped in
whatmann filter paper and placed in 100 ml beaker and selected solvent such as Ethanol,
acetone, hexane and petroleum ether were added in each beaker and kept for overnight. Then
extract was filtered through whatman filter paper and used for the detection of various
phytochemical analyses.

Test for Phytochemicals

 Alkaloids: 2 ml of extract, few drops of Mayers reagents was added, white precipitate
indicates existence of alkaloids.
 Flavonoids: To 2 ml of extract was taken, 10% of lead acetate was added, yellow
precipitate indicate the presence of flavonoids.
 Steroid: 2 ml plant extract was taken in a test tube; 2 ml of CCl3 and few drops
of conc. H2SO4 were added in it and stir well. Chloroform layer appearance red
and acid layer showed the greenish yellow florescence which indicated positive
test for of steroid.
 Glycosides: 3 ml of extract, 4 ml of CCl3 was added and stir well. The chloroform
layer was separated then added 10% ammonium solution. The formation of pink colour
indicate the presence of glycosides.
 Carbohydrates: About 0.5 ml of the extract was taken in which 0.5 ml of Benedict’s
reagent was added. The mixture was heated for 3 minutes in a boiling water bath.
Appearance of red precipitate indicates the presence of carbohydrates.
 Proteins: 2 ml of extract, 2 ml of Millons reagent was added. White precipitates
indicate the presence of proteins.

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 Amino acid: 2 ml of extract, few drops of nitric acid was added. The existance of
yellow colour it indicates the presence of protein and free amino acids.
 Oil: Small quantity of the extract was taken and pressed between two filter papers.
Formation of spot indicates presence of oil.
 Phenol: About 4 ml of extract and 3 ml of 10% lead acetate solution was added .The
formation of bulky white precipitate shows presence of phenol.
 Tannins: 5 ml of extract was taken and few drops of 5% FeCl3 solution was added .The
formation of dark green colour shows presence of tannins.
 Saponins: 0.5ml extract was vigorously shaken with few ml of distilled water. The
formation of forthing is shows presence of saponins.
 Terpenoids: Salkowski’s test- To 4 ml of plant extract, 2 ml of CCl3 and 3ml
Conc. H2SO4 were added carefully to form a layer, reddish brown colour of the
inner face indicate the presence of terpenoids (Thimmaiah, S. R. 1999; Yadav
and Agrwal, 2011; Shailaja J. R. 2015; Sridhar et al., 2016).
Observations and Results:
The phytoconstituents were extracted by using different solvents of increasing polarity
like hexane, acetone, petroleum ether and ethanol. The Table 1 is representing the presence and
absence of different phyto-constituents. Root, stem, leaf and seed extract of selected solvent
revealed the presence of alkaloids, carbohydrate, protein and amino acids. Fruit extract shows
the existence of oil. Glycosides were detected in root, stem, leaf and fruit. Stem extract shoed
the presence of phenol where as other parts showed negative test for phenol. Tannin,
flavonoids, saponins, steroids, terpenoids were detected in all selected parts of C. crista in
different solvents.
Table no: 1. Preliminary phytochemical screening of Caesalpinia crista L.
Sr. Primary Root Stem Leaf Fruit
no. metabolites
E A H P E A H P E A H P E A H P
1. Carbohydrates + + + + + + + + + + + + + + + +
2. Proteins + + + + + + + + + + + + + + + +
3. Amino acid + + + + + + + + + + + + + + + +
4. Oils - - - - - - - - - - - - + + + +

5. Glycosides + - - + - + + - - - + + + + + -
6. Alkaloids + + + + + + + + + + + + + + + +
7. Phenol + - + - - - - - - + + - - + + -
8. Tannins + - - + - - + - + - - + + - - +
9. Flavonoids - + + - + + + + + + + + + + + +
10. Saponins + - + + + + + + + + + + + + + +
11. Steroids + - + - - + - - + - + - + - + -
12. Terpenoids + - - - + + - - - + - + + - - +
(A-Acetone, E-Ethanol, H-Hexane, P- Petroleum ether)

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Conclusion:
It is concluded from the present results that, root, stem, leaf and fruit of C. crista in
different solvent extract revealed the existence of various secondary metabolites. The
medicinal properties such as activity antimalarial activity, antioxidant activity, anti-
amyloidogenic, of burning sensation, convulsions, diabetes wounds, diarrhea, dysentery,
epilepsy, leucorrhea, leprosy, menorrhagia, hemorrhages, ulcers, skin diseases and cytotoxic
activity etc. are due the presence of various secondary metabolites. Hence isolation of
individual compound from the C. crista may found a novel drug.
Acknowledgement:
Authors are greatful to principal of Vidnyan Mahavidyalaya Malkapur, Tq. Malkapur,
Dist. Buldhana – Maharashtra and Bapumiya Sirajoddin Patel Arts, Commerce and Science
College, Pimpalgaon Kale, Tq. Jalgaon (Jamod), Dist. Buldhana – Maharashtra for providing
all the necessary facilities for the present research work.
REFERENCES:
Bhanderi Chandani B., Kalpna D. Rakholiya and Mital J. Kaneria (2022) Pharmacognostic and
phytochemical evaluation of Caesalpinia crista L. leaf. Journal of Medicinal Plants Studies; 10(6): 37-39.
Suryawanshi H. P. and Patel M. R. (2011) Traditional Uses, Medicinal And Phytopharmacological Properties
of Caesalpinia Crista Linn - An Overview, International Journal Of Research In Pharmacy and Chemistry.
Sadiya Afrin, Raihana Pervin, Farah Sabrin , Md. Hossain Sohrab , Satyajit Roy Rony , Md. Emdadul
Islam, Kazi Didarul Islam And Md. Morsaline Billah (2016) Assessment of Antioxidant, Antibacterial and
Preliminary Cytotoxic Activity of Chloroform 0nd Methanol Extracts of Caesalpinia Crista L. Leaf
Biotechnology And Genetic Engineering Discipline, Khulna University, Khulna-9208, Bangladesh. Bangladesh
J. Bot. 45(5): 1061-1068.
Mandal S., Hazra B., Sarkar R., Biswas S. and Mandal N. (2011). Assessment of the antioxidant and reactive
oxygen species scavenging activity of methanolic extract of Caesalpinia crista leaf. Evid. Based Complement.
Alternat. 173768
Srinivas K. V. N. S. Rao Y. K., Mahender I., Das B., Krishna K. V. S. R., Kishore K. H. and Murthy U. S.
(2003) Two new antibacterial flavonoids from the aerial part of Caesalpinia pulcherrima , Phytochemistry,
63:789-793.
Kirtikar K. R. and Basu B. D. (1989) Indian Medicinal Plants, Vol. II, Dehradun: International publishers.
Pooja Upadhyay , Bhuwan Chandra Joshi, Ankush Sundriyal1 and Sushmita Uniyal (2019) Caesalpinia
crista L.: A review on traditional uses, phytochemistry and pharmacological properties, School of Pharmaceutical
Sciences, Sardar Bhagwan Singh University, Balawala, Dehradun, Uttarakhand248001, India yani Inder Singh
Institute of Professional Studies, Dehradun, Uttarakhand-248003, India. Curr Med Drug Res, 3 (1): Article ID
191.
Preedy, V. R., Watson R. R. and Patel V. B. (Eds.). (2011). Nuts and seeds in health and disease prevention.
Academic press.
Thimmaiah, S. R. (1999). Standard methods of biochemical analysis, Kalyani Publishers: 1-435.
Yadav R. N. S., and Agarwala Munin (2011) Phytochemical analysis of some medicinal plants, Journal of
Phytology, 3(12): 10-14.
Shailaja J. R. (2015), Pharmacognostic & Phytochemical Analysis of Gokshura w.s.r. to Cardioprotective
Activity, Anveshana Ayurveda Medical Journal, 1 (5): pp. 391-397.
Sridhar, K., Rajesh, B., Sangeetha, K. (2016), Phytochemical Screening and GC-MS Analysis of Ethanolic
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Remedies, 5(1): 26 – 30.

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29

Removal of Methylene Blue using Cajanus Cajan Activated Carbon

1*Vilayatkar N.D., 1Chaudhari D.L., 2Kadu N.S., 3Kurzekar R.R.

1
S.S.Jaiswal college Arjuni/Morgoan-441701, India
2
Bharatiya college, Amravati - 444605, India
3
C.J.Patel College, Tirora-442201, India
*Corresponding Author; Email: [email protected] Contact No. +91 9028631242

Abstract
Research is required to explore and test new and new low cost materials for removal of
Methylene Blue. In the present research article, generation of activated carbon derived from
Cajanus cajan pericarp, an agricultural waste product, followed by its characterization using
modern techniques like FTIR and SEM studies have been reported. This newly obtained
activated carbon has been tested to estimate its practical applicability for removal of methylene
blue from aqueous solution. Batch adsorption experiments were performed for this purpose.
The influence of pH, contact time, adsorbent doses and initial methylene blue concentration
have also been studied and reported. The results revealed that the activated carbon adsorbent
reported in this article is effective for removal of methylene blue from wastewater, and thus
can be successfully used for control of methylene blue pollution.
Keywords: methylene blue, activated carbon, Cajanus cajan, adsorption isotherm.

Introduction-In water sources occurrence of synthetic dyes gives hazardous effects to aquatic
organism and human health when discharge directly to environment. More than 10,000 dyes
have been widely used in textile, paper, rubber, plastics, leather and cosmetic, pharmaceutical,
and food industries[1].The dyes consist of poisonous and complex components with slow
degradation rate. Furthermore, the presence of dyes components affects the undesirable color
change in water system. The unfavorable impact is not only from esthetics point of view but
also from the decline of sun light penetration, thus reducing photosynthetic activity [2, 3]. As
dyes are designed to resist breakdown with time, exposure to sunlight, soap and oxidizing agent
cannot be easily removed by conventional wastewater treatment processes due to their complex
structure and synthetic origins [4]. MB will cause increased heart rate, vomiting, shock, Heinz
body formation, cyanosis, jaundice, quadriplegia and tissue necrosis in humans [5]. The
commonly used methods for removing metal ions from waste water include precipitation, lime-
coagulation, reduction, electrolytic removal, ion-exchange, reverse osmosis, membrane
filtration, and solvent extraction[6-12]. A number of biosorbent have been used such as tree
barks, saw dusts, activated rice husk, coconut shell, almond shell etc. for the adsorption of
heavy metals. The objective of the present study is to investigate the possible use of activated
carbon derived from Cajanus Cajan pericarp activated Carbon(CCPAC) as an alternative
adsorbent material for removal of MB employing batch experiments.
Materials and Methods-All the chemicals used were of analytical or chemically pure grade.
Distilled water was used throughout the investigation.

Preparation of activated carbon-Cajanus cajan pericarp was collected as an agricultural

waste and washed several times with tap water followed by distilled water. The clean biomass
so obtained was oven dried at 1200C for 24hrs. The dry biomass was milled and sieved to get
fine particles. The biomass was subjected to pyrolysis process using Muffle Furness. During
slow carbonization of raw material in the absence of air at temperature range 700-8000C,
volatile product was removed and residue was converted into char. The char was then subjected

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to chemical activation process using 25% zinc chloride solution. The adsorbent material
obtained has been abbreviated as CCPAC.
Preparation of Methylene Blue solution -A stock solution of methylene blue of concentration
500 ppm (500 mg/L) was prepared by dissolving (0.5000)g of methylene blue in a 1000 mL
volumetric flask, agitated by using a magnetic stirrer for a period of time and completed with
distilled water up to the mark.
Characterization of CCPAC
FTIR analysis: FTIR of CCPAC is presented in Fig.1. The broad band at 3457.44 cm-1
indicates the presence of dissociated or associated –OH groups on the adsorbent surface. The
band at 1637.05cm-1 represents N-H bending. The band at 1119.11cm-1is indicative of CO
stretching. The peak at 618.76cm-1appears to be due to stretching vibration of carbon hydrogen
bonding of alkyne linkages.
Scanning Electron Microscopy (SEM) Analysis: The surface morphology of activated
carbon under investigation was examined using scanning electron microscopy. At x1000
magnification, SEM micrograph clearly revealed that wide varieties of pores are present in
activated carbon along with fibrous structure. It is found that there are holes and caves type
openings on the surface of the adsorbent which would have created more surface area available
for adsorption.

Fig1: FTIR of CCPAC Fig 2: SEM of CCPAC

Results and discussion

Effect of pH-Effect of pH on the adsorption of MB using CCPAC as an adsorbent has been studied
in the pH range 1to10. The relation between the pH of solution and percentage removal of MB has been
shown in fig. 3. The most favorable adsorption was seen at basic pH 5–9 with 70 % removal of MG
dye. So for further study pH 7–8 was maintained.
Effect of Contact time-Fig.4 represents the effect of contact time on removal of MB. The initial MB
concentration taken was 30ppm. The percentage removal has found to increase with time from 35 to
76. After 100 minute negligible effect has been noticed on adsorption of MB. Thus 100 minute is the
optimum time for these batch experiments.
Effect of adsorbent doses-The influence of CCPAC adsorbent doses was studied by varying it from
1 to 10 gm/lit for MB adsorption and the results are represented in Fig.5. The initial MB concentration
taken was 30ppm.The percentage of MB removal was found to increase from 50 to 73. However after
certain dose it becomes constant and it is treated as an optimum adsorbent dose, which is found to be
5gm/lit. for the CCPAC adsorbent.
Effect of Initial MB concentration-In the present study adsorption experiments were performed to
study the effect of initial MB concentration by varying it from 10-100ppm solution is presented in Fig.
6. The results demonstrate that at a fixed adsorbent dose the percentage of MB removal decreases with
increasing concentration of adsorbate.
Adsorption Isotherm-The isotherm data have been linearized using the Langmuir equation and is
plotted between Ce/Qe versus Ce. The Langmuir constant qm, which is measure of the monolayer
adsorption capacity of CCPAC is obtained as 2.85. The Langmuir constant b which denotes adsorption
energy, is found to be 0.315. The high value (0.9122) of regression correlation coefficient (R2) indicates
good agreement between the experimental values and isotherm parameters and also confirms the
monolayer adsorption of MB onto the CCPAC. The dimensional parameter, RL, which is a measure of

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adsorption favorability is found to be 0.0194 (0< RL < 1) which confirms the favorable adsorption
process for MB on CCPAC adsorbent.
75 80
% removal of MB

% removal of MB
65 70
55 60
45 50
35 40
25 30
0 2 4 6 8 10 0 30 60 90 120 150
pH contact time in minute

Fig.3: Effect of pH on adsorption of MB Fig.4: Effect of contact time on MB removal

80 70
% removal of MB

% removal of MB
75 65
70 60
65 55
60 50
55 45
40
50
0 10 20 30 40 50 60 70 80 90 100
0 2 4 6 8 10
Adsorbent dose in gm/lit. Concentration of MB in ppm

Fig. 5: Effect of adsorbent dose on MB adsorption Fig 6: Effect of initial conc. of MB on adsorption

Conclusion-Utilization of waste material such as Cajanus Cajan pericarp for the removal of MB from
the industrial waste-water is investigated. CCPAC is found to be better adsorbent for removal of MB.
The maximum percentage for removal of MB is noticed at pH 7-8 with contact time 100 min. The
percentage removal decrease with increase in initial MB concentration. At 5 gm/lit of optimum
adsorption dose maximum removal efficacy has been noticed. The adsorption data are best fitted with
Langmuir isotherm model which confirms the monolayer adsorption of MB onto CCPAC.
Acknowledgment- Authors are highly thankful to the Principal, S.S.Jaiswal college, Arjuni/Moregaon for
providing necessary laboratory facilities. Authors are also thankful to Director SAIF Punjab University,
Chandigarh and SAIF Cochin University, Kerala for characterization of Carbon.
Reference
[1] S. Mondal, “Methods of dye removal from dye house effluent— an overview,” Environmental Engineering Science, vol.
25, no. 3, 2008, pp. 383–396,.
[2]Spagnoli A A, Giannakoudakis D A and Bashkova S, Adsorption of methylene blue on cashew nut shell based carbons
activated with zinc chloride: The role of surface and structural parameters, Journal of Molecular Liquids 229, 2017, 465–71
[3] Tharaneedhar V, Kumar P S, Saravanan A, Ravikumar C and Jaikumar V. Prediction and interpretation of adsorption
parameters for the sequestration of methylene blue dye from aqueous solution using microwave assisted corncob activated
carbon Sustainable Materials and Technologies 11, 2017, 1–11.
[4]L.Wang, J. Zhang, and A. Wang, “Removal of methylene blue from aqueous solution using chitosan-g-poly(acrylic acid)/
montmorillonite superadsorbent nanocomposite,” Colloids and Surfaces A, vol. 322, no. 1–3, 2008, pp. 47–53.
[5] R. Ahmad and R. Kumar, “Adsorption studies of hazardous malachite green onto treated ginger waste,” Journal of
Environ_mental Management, vol. 91, no. 4, 2010, pp. 1032–1038.
[6] Chakravarti A K , Choudhary S, Chakrabarty S and Mukharjee D C, liquid membrane Multiple emulsion process of
chromium Cr(VI) separation from waste-water colloids, surf A: Physio Chem. Engg. Aspects, 1995, 103, 59-71.
[7] Cimino G, Passerini A and Toscano G, Removal of toxic cations and Cr(VI) from aqeuos solution by hazelnut shell water
Res, 2000, 34(11) 2955-2962.
[8]Gode F and pehlivan E, removal of Cr(VI) from aqueous solution by two lewatit-anion exchange resin, J Hazard Mater,
2005,119 175-182.
[9] Juang RS and Shiau RC , Metal removal from aqueous solution using Chitosan enhanced membrane filtration, J Membr
Sci. 2000, 21(10), 1091-1097.
[10] Lalvani SB, Hubner A and Wiltowski TC, Chromium adsorption in lignin, Energy Sources 2000, 22, 45-46.
[11]Lu A, Zhong S, Chen J, Shi J, Tang J and lu X.Removal of Cr(VI) and Cr(III) from aqueous solution waste water by
natural lino-pyrrhotite Environ Sci. Technol, 2006, 40(9), 3064-3069.
[12]Rengaraj S: Yeon KH and Moon SH, Removal of Chromium from water and waste water by ion exchange resin J. Hazard
Mat. 2001, 87,(1-3) 273-287.

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30
Conductometric Studies of [CPHDD] and [CTMBCD] at Different Molar
Concentrations and Different Percentage Compositions in Water-Ethanol
Mixture
K.P. Jumde1, Sanghapal S. Padhen 2, Saleem R. Khan 3, D.T.Tayade3
1
Department of Chemistry, Nilkanthrao Shinde Science and Arts College, Bhadrawati 442902 (M.S.)India.
2
Department of Chemistry, Rajarshee Shahu Science College, Chandur Railway, Dist. Amravati 444904 (M.S.)
3
Department of Chemistry, Government Vidarbha Institute of Science and Humanities, Amravati,
444604, 3 Department of Chemistry, Government Vidarbha Institute of Science and Humanities,
Amravati, 444604
Abstract
Conductivity play vital role in drug; diffusion, transmission, metabolisum (drug activity and
drug effect) and exerition in pharmokinetics and pharmodynamics of drugs. Thermodynamic
parameters affected by substituent’s of drug. Considering these facts recently in this
laboratory, conductometrically thermodynamic parameters of 4-[4-(4-chlorophenyl]-4-
hydroxypiperidin-1-yl]-N,N-dimethyl-2,2-diphenylbutanamide [CPHDD] and (2S,6R)-7-
chloro-2,4,6-trimethoxy-6'-methyl-3H,4'H-spiro[1-benzofuran-2,1’-]cyclo hex-2-ene]-3,4'-
dione [CTMBCD] were investigated at different molar concentrations and different
percentage compositions. This work mainly highlights on investigation of G, K and µ values.
The thermodynamic parameters viz. ΔH, ΔS and ∆G for ion pair formation were determined
from the value of ion association constant at 298K This technique is suitable and accurate to
study pharmokinetics and pharmodynamics parameters.

Keywords: Conductometric measurements, thermodynamic parameters, [CPHDD],

[CTMBCD] pharmokinetics and pharmodynamics.

Introduction:
Conductivity of drugs/ medicines play key role in medicinal chemistry.
Conductometric measurements along with G, K and µ values, ΔH, ΔS and ∆G and
biopharmaceutical parameters are responsible for the effective bioavailability of drug in vitro
and in vivo correlations [1], which gives useful information regarding the permeability of drug
in drug absorption and transmission of drug in pharmokinetics and pharmodynamics of drug in
drug chemistry. Improvement of solubility and dissolution rate and oral bioavailability of
poorly water soluble drugs are still the challenging aspects for the pharmaceutical technologists
[2]. One of the safest methods of solubilisation is hydrotropic solubilisation [3] and by the
addition of hydrotropic agents the aqueous solubilisation of insoluble drugs can be achieved.
Many researchers highlighted the effect of the solubility enhancers (hydrotropic agents) [4, 5]
and hence improved stability of the drug but no detailed explanation is available relating to the
improvement phenomena.
Split of electrolyte conductivities into the ionic components ideally requires
transference numbers, an accuracy to obtain exact readings for ground level laboratory for
various parameters to chemist become difficult and important task and also serious
experimental problems in many non-aqueous solvents for well equipped laboratories for
pharmacokinetics and pharmacodynamics of drugs in drug chemistry. To overcome this
problem conductance measurements provide very important and valuable information
regarding ion-ion and ion-solvent interactions [6-7]. The conductometric studies of ionic
association of divalent asymmetric electrolyte Cu(NO3)2 with Kryptofix-22 in mixed (MeOH-
DMF) solvents at different temperatures were carried out by Gomaa and Al-Jahdalli [8].

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Izonfuo and Obunwo [9] and Roy et al [10] studied the conductance of alkali metal in different
mixtures mixed solvents. The thermodynamic parameters and Walden products of different
complexes were studied by few researched and they also determined the comparison of
transition metal complexes among the halide groups by [11]. Dubey [12] studied the interaction
between sodium sulphate and 1-propanol, 1-butanol, 1-pentanol and 1-hexanol at different
temperatures by conductometric technique [13]. The present investigation deals with the study
of conductometric properties, thermodynamic behaviour and Walden product of 4-[4-(4-
chlorophenyl]-4-hydroxypiperidin-1-yl]-N, N-dimethyl-2,2-diphenylbutanamide [CPHDD]
and (2S, 6R)-7-chloro -2, 4, 6-trimethoxy-6'-methyl-3H, 4'H-spiro[1-benzofuran 2, 1’-]
cyclohex-2-ene]-3,4'-dione[CTMBCD] in ethanol-water mixture at different concentrations
and different percentage compositions at constant temperature i.e. 270C.
Experimental:
All the chemicals and solvents used for the synthesis were of A.R. grade. All the freshly
prepared solutions were used for investigations. The solvents were purified by standard method
[14]. Different concentration solutions of 0.1M 4-[4-(4-chlorophenyl]-4-hydroxypiperidin-1-
yl]-N,N-dimethyl-2,2-diphenyl- butanamide [CPHDD] and (2S,6R)-7-chloro-2,4,6-
trimethoxy-6'-methyl-3H,4'H-spiro[1-benzofuran-2,1’-]cyclohex-2-ene]-3,4'-
dione[CTMBCD] were prepared. 0.1M solution of each drug was prepared and then by serial
diluton method 0.075M, 0.050M and 0.025M were prepared in 100% water and ethanol-water
mixture respectively. Similar solutions were prepared for 80% and 70% water-ethanol mixture.
All the solutions of drug were always used a fresh in the present investigation.
In 50 ml glass beaker drug solution was taken and it was kept inside the thermostat for
15-20 minutes to attain the thermal equilibrium (270C). After achieving the thermal
equilibrium, the conductivity was measured.
Results and Discussion:
During this investigation conductometric measurements of 100%, 80%, and 70%
mixtures of water-ethanol were prepared. In first set 0.1M solution of [CPHDD] and in second
set 0.1M solution of [CTMBCD] was prepared in conductivity water and by serial dilution
method 0.075M, 0.050M and 0.025M solutions were prepared. At 270C the conductance of
each solution was measured by conductivity bridge and result are cited in Table 1 and Table 2.
From the data observed conductance (G), specific conductance (k) and molar conductance (µ)
were determined by known literature method.

Table 1: Conductometric Measurements of [CPHDD] at different concentration

Determination of G, k and µ on different Concentrations at 27°C
% of solution Concentration Observed Specific Molar
(Water- ethanol) (M) conductance (G) conductance (k) conductance (µ)
100% 0.1 M 0.60X10-3 0.5709606X10-3 5.709606
0.075 M 0.51 X10-3 0.4853165X10-3 6.4708868
0.050 M 0.37 X10-3 0.3520923X10-3 7.0418474
0.025 M 0.21 X10-3 0.1998362X10-3 7.993448
80% 0.1 M 0.73X10-3 0.6946687X10-3 6.9466873
0.075 M 0.70 X10-3 0.6661207X10-3 8.8816093
0.050 M 0.61 X10-3 0.5804766X10-3 11.609532
0.025 M 0.41 X10-3 0.3901564X10-3 15.606256
70% 0.1 M 0.25 X10-3 0.2379002X10-3 2.3790025
0.075 M 0.22 X10-3 0.2093522X10-3 2.7913629
0.050 M 0.20 X10-3 0.1903202X10-3 3.806404
0.025 M 0.14 X10-3 0.1332241X10-3 5.3289656

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Table 2: Conductometric Measurements of [CTMBCD] at different concentration

Determination of G, k and µ on different Concentrations at 27°C
% of solution Concentration Observed Specific Molar
(Water- ethanol) (M) conductance (G) conductance (k) conductance (µ)

100% 0.1 M 0.22X10-3 0.209352 X10-3 2.09352

0.075 M 0.17 X10-3 0.161772 X10-3 2.15696
0.050 M 0.12 X10-3 0.114192X10-3 2.28384
0.025 M 0.08 X10-3 0.076128X10-3 3.04512
80% 0.1 M 0.16X10-3 0.152256X10-3 1.52256
0.075 M 0.15 X10-3 0.14274X10-3 1.9032
0.050 M 0.12 X10-3 0.114192X10-3 2.28384
0.025 M 0.08 X10-3 0.076128X10-3 3.04512
70% 0.1 M 0.15 X10-3 0.14274X10-3 1.4274
0.075 M 0.13 X10-3 0.123708X10-3 1.64944
0.050 M 0.11 X10-3 0.104676X10-3 2.09352
0.025 M 0.07 X10-3 0.066612X10-3 2.66448

From Table-1 to Table-2, it was observed that the observed conductance (G) and
specific conductance (k) were decreases from [CPHDD] to [CTMBCD] continuously while
molar conductance (µ) increases. Decrease in some conductance is due to number of phenolic-
OH group present in the respective molecule. In [CPHDD] observed conductance continuously
decreases from 0.1M concentration to 0.025M concentration continuously. This is due to the
numbers of [CTMBCD] present in these solutions were continuously decreases. Same pattern
was observed in percentage compositions of the mixture. Specific conductance of [CPHDD]
decreases when the molar concentration and percentage composition of water decreases but the
specific conductance increases at the same temperature. In [CPHDD] it was also observed that
molar conductance increases from 0.1M concentration to 0.025M concentration as well as it
increases in all percentage compositions. In 100% water molar conductance is highest while
it will decreases from 100% to 70% water-ethanol percentage compositions. Molar
conductance in 100% water is highest in all molar concentrations. Above results showed that
[CPHDD] and [CTMBCD] are good drugs and will gave good pharmokinetics and
pharmodynamics results of the standard drug. Same patterns of observed conductance, molar
conductance and specific conductance were observed for [CTMBCD]. These results throw
light on pharmokinetics of these two drugs. During this investigation it was observed that the
molar conductance of [CPHDD] is more than [CTMBCD] which clearly indicates the drug
effect of [CPHDD] is comparatively good than [CTMBCD]. It means that the absorption,
transformation and metabolism of [CPHDD] is better than [CTMBCD], so [CPHDD] possesses
best drug activity and drug effect than [CTMBCD].
The specific constant (Ksp), log (Ksp) and thermodynamics parameter viz. change
in free energy (∆G), change in entropy (∆S) and change in enthalpy (∆H) of [CPHDD] and
[CTMBCD] were determined by known literature method at various molar concentration,
percentage compositions and temperatures and result are cited in Table 1 to Table 8.

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Table 3 : Conductometric Measurements of [CPHDD] at different concentration

Determination of Ksp, log Ksp, ∆G,∆H and ∆S at different Concentrations at 27°C
SYSTEM:DRUG CPHDD MEDIUM - 100% WATER
Temp Conc.
Ksp Log Ksp ∆G ∆H ∆S
(°C) (M)
0.100 0.099790372 -1.000911358 5749.37757 -580158.4026 -1953.025934

0.075 0.074842777 -1.125850107 6467.043561 -580158.4026 -1955.418154

27
0.050 0.049895185 -1.301941363 7478.536846 -580158.4026 -1958.789798
0.025 0.024947592 -1.602971367 9207.696116 -580158.4026 -1964.553662

Table 4 : Conductometric Measurements of [CPHDD] at different concentration

Determination of Ksp, log Ksp, ∆G,∆H and ∆S at different Concentrations at 27°C
SYSTEM:DRUG [CPHDD] MEDIUM - 80% WATER
Temp Conc.
Ksp Log Ksp ∆G ∆H ∆S
(°C) (M)
0.100 0.079832297 -1.097821375 6306.042527 -580158.4026 -1954.881484

0.075 0.059874221 -1.222760124 7023.708518 -580158.4026 -1957.233704

27
0.050 0.039916148 -1.398851376 8035.20178 -580158.4026 -1960.645348
0.025 0.019958073 -1.699881393 9764.361124 -580158.4026 -1966.409212

Table 5 : Conductometric Measurements of [CPHDD] at different concentration

Determination of Ksp, log Ksp, ∆G,∆H and ∆S at different Concentrations at 27°C
SYSTEM:DRUG [CPHDD] MEDIUM - 70% WATER
Temp Conc.
Ksp Log Ksp ∆G ∆H ∆S
(°C) (M)
0.100 0.06985326 -1.155813321 6639.156535 -580158.4026 -1955.991864

0.075 0.052389943 -1.280752074 7356.822548 -580158.4026 -1958.384084

27
0.050 0.034926629 -1.456843323 8368.315793 -580158.4026 -1961.755728
0.025 0.017463314 -1.757873327 10097.47506 -580158.4026 -1967.519592

Table 6: Conductometric Measurements of [CTMBCD] at different concentration

Determination of Ksp, log Ksp, ∆G,∆H and ∆S at different Concentrations at 27°C
SYSTEM:DRUG [CTMBCD] MEDIUM - 100% WATER
Temp Conc.
Ksp Log Ksp ∆G ∆H ∆S
(°C) (M)
0.100 3.925965087 0.593946433 -3411.713008 -580158.4026 -1922.488965

0.075 2.944471816 0.469007696 -2694.047086 -580158.4026 -1924.881185

27
0.050 1.962982544 0.292916437 -1682.553784 -580158.4026 -1928.252829
0.025 0.981491271 -0.008113558 46.60543415 -580158.4026 -1934.016693

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Table 7: Conductometric Measurements of [CTMBCD] at different concentration

Determination of Ksp, log Ksp, ∆G,∆H and ∆S at different Concentrations at 27°C
SYSTEM:LIGAND [CTMBCD] MEDIUM - 80% WATER
Temp Conc.
Ksp Log Ksp ∆G ∆H ∆S
(°C) (M)
0.100 3.14077207 0.49703642 -2855.048074 -580158.4026 -1924.344515

0.075 2.355579053 0.372097683 -2137.382152 -580158.4026 -1926.736735

27
0.050 1.570386035 0.196006424 -1125.88885 -580158.4026 -1930.108379
0.025 0.785193016 -0.105023571 603.2703682 -580158.4026 -1935.872243

Table 8 : Conductometric Measurements of [CTMBCD] at different concentration

Determination of Ksp, log Ksp, ∆G,∆H and ∆S at different Concentrations at 27°C
SYSTEM:DRUG [CTMBCD] MEDIUM - 70% WATER
Temp Conc.
Ksp Log Ksp ∆G ∆H ∆S
(°C) (M)
0.100 2.748175561 0.439044473 -2521.934061 -580158.4026 -1925.454895

0.075 2.061131671 0.314105736 -1804.268139 -580158.4026 -1927.847115

27
0.050 1.374087781 0.138014477 -792.7748368 -580158.4026 -1931.218759
0.025 0.687043889 -0.687043889 936.3843814 -580158.4026 -1936.982623
From Table-3 to Table-8 it was observed for these drugs Ksp, log Ksp, ∆H and ∆S
decreases continuously while ∆G increases when we go from 0.1M concentration solution to
0.025M concentration. Same pattern was observed in percentage composition of the mixture
i.e. these thermodynamic parameters are highest in 100% water while least in 70% water-
ethanol solvent. In [CTMBCD] the values of all thermodynamic parameter as well as Ksp and
log Ksp are the greatest than [CPHDD].
Conclusion:
From this investigation it is clear that various functional groups such as electron
donating, electron withdrawing, acidic, basic and various functional groups present in the
molecule directly affect conductance, specific conductance, molar conductance, Ksp, ∆H, ∆S
and ∆G values of that drug. The structure of the drug as well as nature of that drug directly
affects these parameters. The temperature molar concentrations and percentage compositions
are also responsible for changing the values of these parameters. The solute (drug)-solvent
interactions, solvent-solvent interactions, solvent-solvent-solute interactions and solute-solute-
solvent interactions are another factor which directly hamper these parameters. The internal
geometry as well as internal and intra hydrogen bonding affect these parameters.
During this investigation it was also observed that the molar conductance of [CPHDD]
is highest than [CTMBCD] which clearly indicates the drug effect of [CPHDD] is
comparatively more than [CTMBCD].
Disclosure of conflict of interest:
Authors wish to state that there is no conflict of interest on this work.

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References:
1 S.Chakraborty, D.Shukla, A.Jain,B.Mishra and S.Singh, J.Coll.Int.Sci.,355,242-249,(2009).
2 S.Agrawal,S.S.Pancholi, N.K.Jainand G.P.Agrawal, Int.J.PHam.,274,149-155,(2004).
3 G.D.Pancholi and J.C.Gradock, J.PHarm.Sci.,68,728-732,(1974).
4 U.N.Dash and S.Supkar, Proc.Ind.Acad.Sci.Chem.Soc.,107, 541,(1995).
5 Agnieszka Boruń, Conductance and ionic association of selected imidazolium ionic liquids in
various solvents: A review, Journal of Molecular Liquids, 276,2019, 214-224,
https://doi.org/10.1016/j.molliq.2018.11.140.
6 E. A. Gomaa, B. M. Al-Jahdalli, Conductometric Studies of Ionic Association of Divalent
Asymmetric Electrolyte Cu(NO3)2 with Kryptofix -22 in Mixed (MeOH-DMF) Solvents at
Different Temperatures, American Journal of Condensed Matter Physics, Vol. 2 No. 1, 2012,
pp. 16-21. doi: 10.5923/j.ajcmp.20120201.03.
7 W.A.L.Izonfuo, C.C.Obunwa, Ind.J.Chem.,38A,939,(1999).
8 M.N.Roy, D.Nandi, and D.K.Hazra, J.Ind.Chem.Soc.,70,121,(1993).
9 G.C.Bag, N.M.Singh, N.R.Singh, J.Ind.Chem.Soc.,77,146,(2000).
10 N.Dubey, J. Surface Sci. Technol., 24, 139-148, (2008).
11 A.B. Wadekar , D. T. Tayade, [2016], Conductometric Study of
Substitutedthiocarbamidonaphthols in 70% Ethanol– Water Mixture at Different Molar
Concentrations at Constant Temperature, International Journal of Science and Research
5(1),678-680
12 Agnieszka Boruń, Conductance and ionic association of selected imidazolium ionic liquids in
various solvents: A review, Journal of Molecular Liquids, 276,2019,214-224,
https://doi.org/10.1016/j.molliq.2018.11.140.
13 Bahram Ghalami-Choobar, Tayyebe Nosrati Fallahkar, [2019], Thermophysical properties of
1-ethyl-3-methylimidazolium bromide ionic liquid in water + ethylene carbonate mixtures at
T = (298.2, 308.2 and 318.2) K Fluid Phase Equilibria, Volume 496, 42-60
14 A. B. Wadekar , D. T. Tayade, [2016], Thermodynamic study of substituted thiocarbamido-
Napthonols at different concentrations and different Temperatures in mixed solvent media”
European Journal of Pharmaceutical and Medicinal Research, 3(9), 635-637

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31
Synthesis and Characterization of Some Substituted Chalcone by the
Green Synthesis Way (Grinding Method)
Nilesh S. Padole1, Murlidhar P. Wadekar2, Shubhangi Y. Deshmukh3,
Vinod M. Sherekar4
1
Head and Assistant Professor, Vinayak Vidnyan Mahavidyalaya Nandgaon Khandeshwar 444708.
2
Associate Professor, GVISH, Amravati, 444602.
3
Reseacrh student, VBMV Camp, Amravati 444602.
4
Assistant Professor, Vinayak Vidnyan Mahavidyalaya Nandgaon Khandeshwar 444708.
*Corresponding Author: Name: Mr. Nilesh S. Padole, E-mail: [email protected] Phone: +91-
9096312784, Address: Vinayak Vidnyan Mahavidyalaya, Nandgaon Khandeshwar, 444708.

ABSTRACT
The presented chalcone were synthesized by Claisen- Schmidt condensation of 2-hydroxy-3-
nitro-5-methyl acetophenone (3) and o-chloro benzaldehyde. m-chloro and indol-3-carboxy
aldehydes. All the synthesized compounds were characterized through NMR, IR, and elemental
analysis. The synthetic methods used proved successful synthesis of the desired compound.
Grinding technique is used to follow the green synthesis way for the synthesis of chalcone.
KEYWORDS: Chalcones, Green Synthesis (Grinding Technique), Claisen- Schmidt
condensation.
INTRODUCTION
Alpha-beta unsaturated ketones or commonly referred as chalcones are one of the
important classes of organic compounds frequently encountered in synthetic chemistry. They
are important intermediates not only as key building blocks for the synthesis of core
heterocycles such as pyrazole, isoxazole, triazole, flavone, benzodiazepine and pyrimidine in
medicinal chemistry, but also as an invaluable chelating ligand for various lanthanide and
transition metals in material chemistry. Chalcones, α, β‐unsaturated carbonyls, play a crucial
role in many biological processes as well as are well-known intermediates for synthesizing
various heterocycles1 and bear a very good synthon so that variety of novel heterocycles with
good pharmaceutical profile can be designed2. Presence of enone functionality in chalcone
shows the antibiotic activity.
Recent trends in organic synthesis utilize a non-conventional green technique such as
ultrasound (sonochemistry), microwave irradiation, grinding and by using ionic liquids which
have also been proved to have many advantages3,4,5 Numerous solvents assisted and solvent-
free methods have been reported for the preparation of pyrazoline derivatives. The present
article elaborates on various green techniques reported previously for synthesis of chalcone
derivatives, which will be useful for researchers for synthesizing chalcone derivatives in less
time, yield effective and safely.
Green Synthesis of chalcone Derivatives “The design of chemical products and
processes that are more environmentally benign and reduce negative impacts to human health
and the environment”6. There are various green techniques available in the literature for
synthesis of chalcone derivatives. Some green methods for organic synthesis of pyrazolines7,8
Microwave Irradiation Ultrasonic Irradiation Grinding Technique Ionic Liquids Microwave
Irradiation Microwave, which consists of electric and magnetic field, acts as a non-ionising
reaction that causes ionic motions and rotations, but there is no change in molecular structure.
At least one liquid polar is required in microwave synthesis9. Polar mechanism shows the

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microwave effect that when polarity is increased from ground state towards the transition state
during the reaction10.
EXPERIMENTAL SECTION
The chalcones were synthesized by using 2-hydroxy-3-nitro-5-methyl acetophenone
with some aldehydes (2-chloro Benzaldehyde, 3-chloro Benzaldehyde and Indol -3-
carboxaldehyde) under alkaline condition. The chalcones were obtained yellow to brown and
peach in colour and their synthesis were confirm by melting point method and from spectral
interpretation. This is the general Claisen-Schmidt reaction11 but I used the reaction by
grinding was carried out in a mortar and pestle at room temperature. During the grinding,
sudden change in colour took place, indicating the progress of reaction. The full conversion of
the reaction mixture to a solid mass indicates the completion of reaction12
The chemicals and solvents used were of highest purity purchased commercially from
Merck, S.D. Fine and Alfa Aesar Company Ltd. The melting points of all the synthesized
compounds were recorded by Thiele’s melting point apparatus as uncorrected values. The
elemental analysis was carried out on Thermo Scientific CHNS elemental analyser. IR spectra
were recorded on a Shimadzu instrument using KBr pallet. 1H NMR spectra were scanned by
Brucker at 400 MHz using DMSO-d6 as solvent and TMS as an internal reference. 13C NMR
spectrum of one sample (4a) was recorded on same instrument at 100 MHz. Experimental
procedure for synthesis of 3-(substituted)-1-(2-hydroxy-5-methyl-3-Nitro phenyl) prop-2-en-
1-ones 5(a-c).
SYNTHESIS OF CHALCONES
Preparation of p-methylphenyl acetate (1)
The p-cresol was refluxed along with acetic anhydride and anhydrous sodium acetate for an
hour. The reaction mixture was cooled and poured into the ice-cold water containing crushed
ice. Acetate layer was separated by means of separating funnel and several times washed with
water. It was finally purified by distillation and the distillate fraction was collected at about
2360C, to get the compound (1) b.p. 2360C yield: 84.74%.
Preparation of 2-hydroxy-5-methyl acetophenone (2)
p-methyl phenyl acetate (1) was mixed with anhydrous AlCl3 (1) and heated at 1200C for
45minutes on an oil bath. The reaction mixture was decomposed in ice cold water containing
10% hydrochloric acid and allowing the solution to fall drop by drop into ice cold water with
constant stirring. Green solid compound i. e crude ketone (2) was obtained, m.p. 470C, yield:
89%.
Preparation of 2-hydroxy-3-nitro-5-methyl acetophenone (3):
2-hydroxy-5-methyl acetophenone (2) was dissolved in acetic anhydride in a beaker and
reaction mixture was kept in ice bath by maintain temperature below 50C. To this reaction
mixture conc. HNO3 was added dropwise with constant stirring till the solution becomes orange
coloured and kept for 4-5 hrs. it was then decomposed with ice cold water. Yellow granules
obtained were filtered and washed with water and then crystallized from ethanol, m.p. yield:
72%.
Preparation of β-unsaturated chalcones (4a–4c)
A mixture of 2-hydroxy-3-nitro-5-methyl acetophenone (3) (0.01mol) and chloro substituted
aldehyde and indole 3-carboxyaldehyde, (0.01mol), aryl and ammonium bromide and

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ammonium persulphate moist with few drops of water is ground at room temperature for 25
minutes in a mortar and pestle. The reaction mixture is allowed to stand for 25 min. Then Ice-
cold water (30 ml) was added to the reaction mixture and acidified with conc. HCl. The product
was collected by vacuum filtration and then recrystallized from ethanol. The same procedure
of grinding was applied by using anhydrous Barium hydroxide and then result of two procedure
was compared. Name of the chalcone prepared are entitled below.
 3-(2-chlorophenyl)-1-(2-hydroxy-5-methyl-3-nitrophenyl)prop-2-en-1-one (4a)
M.P. 1120C, yield: 74%.
 3-(3-chlorophenyl)-1-(2-hydroxy-5-methyl-3-nitrophenyl)prop-2-en-1-one (4b)
M.P 1180C, yield: 84%.
 1-(2-hydroxy-5-methyl-3-nitrophenyl)-3-(1H-indol-3-yl)prop-2-en-1-one (4c) M.P
1540C, yield: 76%.
The complete experimental scheme for synthesis of above titled compounds is given below.

Scheme of 3-(substituted)-1-(2-hydroxy-5-methyl-3-Nitro phenyl) prop-2-en-1-ones 4(a-

c). R1= NO2; R2=2-Chloro benzaldehyde, 3- Chloro benzaldehyde, Indol-3-carboxyaldehyde,
RESULTS AND DISCUSSION
Spectroscopic Data: The IR, 1H NMR and 13C NMR spectral data showed expected signals or
peaks which correspond to various groups present in each compound. Also, elemental analysis
was found in full agreement with the proposed structures. The elemental analysis, IR, 1H NMR
and 13C NMR spectral data of compounds 4(a-c) are shown below
3-(2-chlorophenyl)-1-(2-hydroxy-5-methyl-3-nitrophenyl)prop-2-en-1-one (4a)
Brown solid; M.P. 1120C, yield: 74%.; Elemental Anal. Calcd. for C16H12ClNO4.: C: 60.49, H:
3.81, O:4.41, Cl: 11.16 Found: C: 53.10, H: 2.40, O:3.98, Cl: 21.00. IR (KBr) cm-1: 3120
(Phenolic OH stretch), 2980 (Aromatic C-H stretch), 2850 (Aliphatic C-H stretch), 1695 (C=O
stretch), 1520 (Aromatic C=C stretch),).1H NMR (400 MHz, DMSO-d6) δ (ppm): 1.51 (s, 3H,
-CH3), 3.43 (s, 2H, -CH2), 3.80 (s, 1H, -OH), 8.15-8.32 (m, 6H, Ar-H).13C NMR (100MHz,
DMSO-d6) δ (ppm): 40 (-CH2), , 195 (C=O), 123-136 (Ar-C), 149- 165 (C=C).
3-(3-chlorophenyl)-1-(2-hydroxy-5-methyl-3-nitrophenyl)prop-2-en-1-one (4b)
Dark brown solid; M.P 1180C, yield: 84%.; Elemental Anal. Calcd. for C16H12ClNO4.; C:53.10;
H:2.40; N:3.98, Cl: 21.00. Found: C: 60.49; H: 3.81; N: 4.41, Cl:20.91. IR (KBr) cm-1: 3340
(Phenolic OH stretch), 2980 (Aromatic C-H stretch), 2910 (Aliphatic C-H stretch), 1615 (C=O
stretch), 1515 (Aromatic C=C stretch), 1260 (C-O stretch).1H NMR (400MHz, DMSO-d6) δ
(ppm): 1.51 (s, 3H, -CH3), 3.81 (s, 2H, -CH2), 5.13 (s, 1H, -OH), 6.99-7.90 (m, 6H, Ar-H).
1-(2-hydroxy-5-methyl-3-nitrophenyl)-3-(1H-indol-3-yl)prop-2-en-1-one (4c)

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Peach color solid; M.P 1540C, yield: 76%; Elemental Anal. Calcd. for C18H14N2O4 : C, 67.08;
H: 4.38; N: 3.98, found C: 53.10; H: 3.40; N: 7.90. 1H NMR (400MHz, DMSO-d6) δ (ppm):
1.50 (s, 3H, -CH3), 3.38 (s, 2H, -CH), 4.10 (s, 1H, -OH), 6.77-7.65 (m, 6H, Ar-H).
CONCLUSION
In conclusion, a new series of 3-(substituted)-1-(2-hydroxy-5-methyl-3-Nitro phenyl) prop-2-
en-1-ones i.e chalcones 4(a-c) bearing 4-methyl phenol i.e. p-cresol moiety were successfully
synthesized in satisfactory yield by employing Claisen-Schmidt condensation of corresponding
substituted 2-hydroxy-3-nitro-5-methyl acetophenone (3) and their structures were elucidated
by chemical characteristics, elemental analysis and IR, 1H NMR and 13C NMR spectroscopic
techniques.
ACKNOWLEDGEMENT
Authors are thankful to Research centre of department of Chemistry, Government Vidharbha
Institute of Science and Humanities, Amravati for providing research facilities.
REFERANCES
1
. Khan SA, Asiri AM, Kumar S, Sharma K. European Journal of Chemistry. 2014 Mar
31;5(1):85-90. 7
2
. Al-Bogami AS, Alkhathlan HZ, Saleh TS. 2013 Jul 21;25(11):6427
3
. Deshmukh S.Y. Padole N.S. , Wadekar M.P. , Chaudhari M.A.; Journal of Chemical and
Pharmaceutical Research, 2021, 13(7):01-06
4
. . ElShora AI. Egypt J Sol. 2000;23:251-4.
5
. Shubhangi Y. Deshmukh1*, Nilesh S. Padole2; JETIR,2022,9(2): 794-801
6
. Anastas PT, Warner JC. Principles of green chemistry. Green chemistry: Theory and
practice. 1998:29-56.
7
. Thirunarayanan G, Mayavel P, Thirumurthy K, Kumar SD, Sasikala R, Nisha P,
Nithyaranjani A.. European Chemical Bulletin. 2013 Apr 18;2(9):598-605.
8
. Chemat F, Esveld E. Chemical engineering & technology. 2001 Jul 1;24(7):735-44
9
. Bougrin K, Loupy A, Soufiaoui M. Journal of Photochemistry and Photobiology C:
Photochemistry Reviews. 2005 Oct 31;6(2):139-
10
. Perreux L, Loupy A. Tetrahedron. 2001 Nov 5;57(45):9199-223.
11
. Shubhangi Y Deshmukh, IJFMR,23-212,1-6, (2023).
12
. K.P. Kakade, S. P. Kakade and S Y. Deshmukh, world journal of pharmacy and
pharmaceutical science,4,1,1591-1597 (2014).

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32

In-Silico Prediction of Phytoconstituents From

Solanum Indicum for Antiepileptic Activity

Pooja.P.Patle*, Parimal Katolkar, Pradeep Raghatate, Jagdish Baheti

Kamla Nehru College of Pharmacy, Butibori, Nagpur 441108 (MS), INDIA
*Email: [email protected]

ABSTRACT
Objective
A persistent, non-communicable brain disorder is epilepsy. Its distinctive characteristic is
recurrent seizures. One or more parts of the body may experience partial or generalised
seizures, which are short bursts of uncontrollable movement that can occasionally be
followed by loss of consciousness and control over bowel or bladder function. Compounds
found in medicinal plants have been the source of many conventional medications. In-silico
testing of Solanum indicum phytoconstituents for antiepileptic efficacy was a part of our
investigation.
Methods
Utilizing Discovery studio, molecular docking is done to assess the pattern of interaction
between the phytoconstituents from the Solanum indicum plant and the crystal structure of
the epilepsy proteins (PDB ID: 6O4L). Later, SwissADME and pkCSM were used to screen
for toxicity as well as the pharmacokinetic profile. Results
The docked results suggest that Solafuranone (-7.8 kcal/mol), Isofraxidin (-6.1 kcal/mol) for
6O4L macromolecule has best binding towards antiepileptic activity as compared to the
standard (Acetazolamide) for 6O4L is -5.1 kcal/mol. Furthermore, pharmacokinetics and
toxicity parameters were within acceptable limits according to ADMET studies.
Conclusion
Results from the binding potential of phytoconstituents aimed at antiepileptic activity were
encouraging. It promotes the usage of Solanum indicum and offers crucial details on
pharmaceutical research and clinical care.
1. INTRODUCTION
Solanum indicum, also known as Birhata, Badi Kateri, or Indian night shade, belongs to the
Solanaceae family. It is a common upright undershrub in warmer parts of India, Asia, and
Africa that grows up to 1.5 metres in height. Its height ranges from 0.30 to 1.8 metres.
Nationally, 500–1000 MT are needed each year. The berries, leaves, roots, seeds, and stems
of this plant have all been used in traditional medical systems to treat a wide range of
ailments, including bronchitis, asthma, dry cough, rhinitis, dysuria, leucoderma, sexual
dysfunction, insomnia, heart weakness, and pruritis.1
Solanum indicum, which is consumed as a vegetable in some regions of Africa, may prevent
cardiovascular diseases. It was exciting to read the studies on how it may be used to treat
hypertension. The extract's potent therapeutic and preventative benefits against hypertension
may not have been possible without the presence
of chlorogenic acids. The antihypertensive effect might be due to other components. The
findings urge further research into the extract as a possible treatment for hypertension.2
However, there are few studies on the phytoconstituents of Solanum indicum for the

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antiepileptic activity. Thus, keeping the above information in view, the present investigation
was designed to identify the potential phytochemicals of Solanum indicum against 6O4L
using a molecular docking method.
2. MATERIALS AND METHODS
2.1. Platform for molecular docking
The computational docking study of all the phytoconstituents selected as ligands with
antiepileptic activity as the target was performed using PyRx software.3
2.2. Protein preparation
The macromolecule is 6O4L, in silico analysis of selected phytoconstituents was performed
on the 1.85 Å crystal structure of antiepileptic macromolecule with inhibitor, (PDB ID: 6O4L,
having resolution Resolution: 1.85 Å, R-Value Free: 0.220, R-Value Work: 0.174, R-Value
Observed: 0.176), which was retrieved from the protein data bank (https://www.rcsb.org).
6O4L is classified as Crystal Structure of ALDH7A1 mutant E399D complexed with NAD all
other molecules, such as co-crystallized water molecules, unwanted chains, and nonstandard
residues, were deleted. Using Discovery studio.4
2.3. Mechanism of Action
6O4L: Aldehyde dehydrogenase 7A1 (ALDH7A1) is an enzyme that catabolizes lysine,
and some loss-of- function mutations in this gene result in pyridoxine-dependent epilepsy
(PDE). To understand how the mutations affect NAD+ binding, the crystal structures of the
mutant enzymes in complex with NAD+ were examined. At the tested dose of NAD+, these
alterations result in less active tetrameric ALDH7A1.5
2.4. Ligand preparation
The three-dimensional (3D) structures of all constituents were retrieved using Avogadro
software from the PubChem database available on the NCBI website
(https://pubchem.ncbi.nlm.nih.gov/). However, the drawing of geometrical 2D structure was
performed using the ChemSketch program. The two-dimensional (2D) structures were
transformed into 3D models using the Avogadro software and the ligand structures were
saved in the PDB format. All the chemical structures are shown in Figure 1.

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2.5. Standard Preparation

The standard is prepared steps such as, the 2D structure of standard drug was made using chem
sketch program, then the 2D structure was converted into 3D model using Avogadro
Software, it was saved in PDB format.
By using PyRx molecular docking of Acetazolamide was done with 6O4L.
2.6. Molecular docking
Molecular docking evaluates the protein-ligand interactions and estimates the scoring
function based on the geometry to predict the binding affinity of the ligand molecule6,7. We
applied molecular docking studies to investigate the binding pattern of selected
phytoconstituents (Figure 1) and the standard drug, along with the crystal structure of
antiepileptic activity macromolecule (PDB ID: 6O4L). The molecular docking study was
performed using PyRx software, Binding affinity was explored using the Vina wizard tool.
The final results were analysed and visualized using Discovery Studio 2020 Client 8, with
bound ligands as the standard. Visualization of protein ligand interaction reflects the number
of interactions and active residues responsible for significant binding at the active site of the
target enzyme.
2.7. Absorption, distribution, metabolism, and excretion (ADME) and toxicity prediction
The selected phytoconstituents and standard drug were further checked for drug-likeness
properties according to Lipinski’s rule. During drug development, it is necessary to predict the
tolerability of phytochemicals before being ingested by humans and animal models. The
pharmacokinetic profile (ADME) and toxicity predictions of ligands were conducted using
SwissADME (http://www.swissadme.ch) and pkCSM (an online server database
predicting small-molecule pharmacokinetic properties using graph-based
signatures, (http://biosig.unimelb.edu.au/pkcsm/prediction). To analyse the toxicological
properties of ligands, Simplified Molecular Input Line Entry System (SMILES) notations or
PDB files were uploaded, followed by selecting the required models for generating
numerous information about structure-related effects9,10.
3. RESULT AND DISCUSSION
The present study aimed to explore the inhibitory potential of the phytoconstituents present
in Solanum indicum targeting antiepileptic activity. In this study, we performed molecular
docking studies of all phytoconstituents found in Solanum indicum using AutoDock Vina,
followed by a study of interacting amino acid residues and their influence on the inhibitory
potentials of the active constituents. Selected phytoconstituents showing the best fit were
further evaluated for absorption, distribution, metabolism, excretion, and toxicological
(ADMET) properties using SwissADME and pkCSM servers.
3.1 Molecular docking
The docking scores and binding energies of all chemical constituents of Solanum indicum
targeting antiepileptic activity (PDB ID: 6O4L) and binding interactions with amino acid
residues are presented in Table 1 respectively.
Table 1. Binding interaction of ligands from Solanum indicum targeting antiepileptic
activity (PDB ID:6O4L)

Docking Score
Sr. No. Chemical constituent PubChem ID
6O4L
1. Solanidine 65727 -10.0

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2. Solasodine 5250 -9.0

3. Solavetivone 442399 -7.3
4. Solafuranone 11107208 -7.8
5. Scopoletin 5280460 -6.5
N-p-trans-
6. 5372945 -6.7
Coumaroyltyramine
7. Isofraxidin 5318565 -6.1
8. Lauric acid 3893 -3.7
9. 𝛽-sitosterol 222284 -7.1
10. Diosgenin 99474 -8.1
Standard Drug
11 Acetazolamide 1986 -5.1

The binding affinities of phytoconstituents ranged from –10.0 to -3.7 kcal/mol. From the
docked results, it is evident that the compounds, Solanidine exhibit the most favourable
binding affinity (−10.0 kcal/mol) in complex with selected macromolecules (PDB ID: 6O4L)
as compared to other docked compounds i.e., Solsodine (-9.0 kcal/mol), Diosgenin (-8.1
kcal/mol), Solafuranone (−7.8. kcal/mol), Solavetivone (−7.3 kcal/mol), Beta sitosterol (-7.1
kcal/mol), N-p-trans-Coumaroyltyramine (−6.7 kcal/mol), Scopoletin (−6.5 kcal/mol),
Isofraxidin (−6.1 kcal/mol), Lauric acid (−3.7 kcal/mol). Visual examination of the
computationally docked optimal binding poses of phytoconstituents on selected
macromolecules (i.e., 6O4L) revealed the significant involvement of various types of
interactions, such as hydrogen bonding and hydrophobic interactions, including π–π stacking
and π–alkyl and alkyl interactions, in the stability of the binding of the phytoconstituents to
6O4L.
The binding affinity of the standard (Acetazolamide) for 6O4L is -5.1 kcal/mol.
3.1.1. Solafuranone, 6O4L
The number of intermolecular hydrogen bonds, the binding energy of ligand 6O4L stable
complexes, and the number of nearest amino acid residues were also determined for selected
compound Solafuranone All synthesized derivatives formed complexes with target proteins.
Analysis of interactions of the 6O4L protein complex and ligand Solafuranone showed that
the ligand molecule is oriented due to Conventional Hydrogen
bond with THR A: 346 amino acid residue and Pi-Pi Stacked interaction with PHE A: 401
amino acid residue were found.
3.1.2. Isofraxidin, 6O4L
An analysis of the interactions between the 6O4L protein complex and the Isofraxidin ligand
was also carried out, which showed that the ligand molecule is oriented due to conventional
hydrogen bond with the ASP A: 399, THR A: 346, PHE A: 401 amino acid residue and Pi-
Alkyl interaction with ALA A: 349 amino acid residue were found.
3.1.3. Acetazolamide, 6O4L
The binding affinity of the standard (Acetazolamide) for 6O4L is -5.1 kcal/mol. the

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interactions between the 6O4L protein complex and the Acetazolamide ligand was also
carried out, which showed that the ligand molecule is oriented due to conventional hydrogen
bond with the PHE A:166, THR A:164 amino acid residue, Carbon hydrogen bond with PRO
A:193 amino acid residue, Pi-Sulfur interaction with PHE A:166 amino acid residue, Pi-
Cation interaction with LYS A:190 and Pi-Alkyl interaction with ALA A:165, ALA A:192
amino acid residue were found.
Table No. 2. Interactions with amino acid residue.

Binding Main amino acid interactions

Sr. Energy Pi-alkyl, alkyl, Pi-S/Pi-Pi, Van der Waals
No. Molecule (kcal/mol) H bond T interactions
shaped/halogen/unfavoura
ble
donor-donor
interactions
1 Solanidine -10.0 No ALA A:192, ALA A: 165, No
interactions VAL A: 250 interactions
2 Solasodine -9.0 GLY A: VAL A: 250, PRO A: 193 No
226 interactions
ASN A: No
3 Solavetivone -7.3 379, ASN TYR A: 41 interactions
A: 46
4 Solafuranone -7.8 THR A: PHE A: 401 No
346 interactions
THR A: No
5 Scopoletin -6.5 164, PHE CYS A: 302, ALA A: 165 interactions
A: 401
6 N-p-trans- -6.7 SER A: PHE A: 168, CYS A: No
Coumaroyltyram 460 302, interactions
ine TRP A: 175, PHE A:
468
7 Isofraxidin -6.1 ASP A: ALA A: 349 No
399, PHE interactions
PHE A: PRO A: 288, LEU A: 285, No
8 Lauric acid -3.7
292 PRO A: 458 interactions
TRP A: 31, ALA A: 207,
No LYS No
9 𝛽-sitosterol -7.1 interactions interactions
A: 204, LYS A: 208,
VAL A: 209,
ALA A: 92
ASN A: No
10 Diosgenin -8.1 TYR A: 41, PHE A: 166
379 interactions
THR A:
164, PHE
A: 166, PRO ALA A: 165, ALA A: 192, No
11 Acetazolamide -5.1 A: interactions
193 LYS A: 190

3.2. ADMET study

Pharmacokinetic profile (ADME) and toxicity predictions of the ligands are important
attentive parameters during the transformation of a molecule into a potent drug. In the present
study, these parameters were assessed using SwissADME and pkCSM. The absorption
potential and lipophilicity are characterized by the partition coefficient (Log P) and
topological polar surface area (TPSA), respectively. For better penetration of a drug

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molecule into a cell membrane, the TPSA should be less than 140 Å. However, the value of
Log P differs based on the drug target. The ideal Log P value for various drugs are as follows:
oral and intestinal absorption,
1.35 − 1.80; sublingual absorption, > 5; and central nervous system (CNS)11. The aqueous
solubility of ligands ideally ranges from − 6.5 to 0.5 12, while the blood brain barrier (BBB)
value ranges between − 3.0 and 1.2 13. In addition, non-substrate P-glycoprotein causes drug
resistance 14.
In our study, all the selected ligands followed the TPSA parameter, P-glycoprotein non-
inhibition, thereby showing good intestinal absorption and an acceptable range of BBB
values. All the compounds showed aqueous solubility values within the range. Further, it
was predicted that the selected ligands do not show AMES toxicity, hepatotoxicity, and skin
sensitivity. In addition, it did not inhibit hERG-I (low risk of cardiac toxicity). Lipinski’s rule
violations, T. pyriformis toxicity, minnow toxicity, maximum tolerated dose, rat acute oral
toxicity, and chronic toxicity are depicted in table.
Table 3. ADME and toxicity predicted profile of ligands with superior docking score

Aqueou s Human Blood

MW HB Hb accept solubili ty intestina l Brain
ADMET Formula (g/mol Log P TPSA (Ȧ2) done r or (Log absorpti Barri er
Properties ) mol/L) on (%)
Solanidine 397.64 5.655 23.47 1 2 -4.927 92.975 0.695
C27H43NO 7 Ȧ2
Solasodine 413.64 5.2869 41.49 2 3 -3.809 92.324 0.035
C7H43NO2 Ȧ2
Solavetivone 218.33 3.9042 17.07 0 1 -4.615 95.873 0.635
C15H22O Ȧ2
Solafuranone 232.32 3.1876 26.30 0 2 -3.551 95.523 0.206
C15H20O2 3 4 Ȧ2
Scopoletin 192.17 1.5072 59.67 1 4 -2.504 95.277 -0.299
C10H8O4 Ȧ2
N-p-trans- 283.32 69.56
Coumaroyltyram C17H17NO3 7 2.4699 Ȧ2 3 3 -3.165 90.031 -0.552
ine
Isofraxidin 222.19 1.5158 68.90 1 5 -2.458 95.588 -0.377
C11H10O5 6 Ȧ2
Lauric acid 200.32 3.9919 37.30 1 1 -4.181 93.379 0.057
C12H24O2 2 Ȧ2
𝛽-sitosterol 414.71 8.0248 20.23 1 1 -6.773 94.464 0.781
C29H50O 8 Ȧ2
Diosgenin 414.63 5.7139 38.69 1 3 -5.713 96.565 0.2
C27H42O3 Ȧ2
Acetazolamide C4H6N4O3 222.25 - 151.66 2 6 -2.428 59.043 -0.622
S2 1 0.8561 Ȧ2

Table 3 Continued

Total Max
P- clearanc tolerated hERG
glyco- Bioavailabili AMES hER II
ADMET e [Log dose
protein ty score GI inhibit
mL/ toxicit [Log
Properties substrat score or
(min.kg y mg/ inhibi
e )] (kg.d)] to r
Solanidine YES 0.028 0.55 NO -0.882 NO YES

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Solasodine YES 0.09 0.55 NO -0.375 NO YES

Solavetivone NO 1.225 0.55 NO 0.044 NO NO
Solafuranone NO 1.256 0.55 NO 0.526 NO NO
Scopoletin NO 0.73 0.55 NO 0.614 NO NO
N-p-trans-
Coumaroyltyrami
YES 0.265 0.55 NO -0.213 NO YES
e
Isofraxidin NO 0.713 0.55 NO 0.56 NO NO
Lauric acid NO 1.623 0.85 NO -0.34 NO NO
𝛽-sitosterol NO 0.628 0.55 NO -0.621 NO YES
Diosgenin NO 0.328 0.55 NO -0.559 NO YES
Acetazolamide NO -0.01 0.55 NO 1.263 NO NO

Table 3 Continued

Oral rat Minno

chronic T.Pyrifor w
Acute oral mis Lipinski
toxicity Skin toxicit
rat ’s rule
ADMET (Log Hepatotoxic sensati toxicity y (Log
toxicity. (Log Violatio
mg/kgbw/d ity on mmol/
Properties LD50(mol/ μg/L) n
ay) L)
kg)

Solanidine 2.596 1.334 YES NO 0.378 -0.493 YES (1)

Solasodine 2.489 1.332 YES NO 0.311 0.381 YES (1)

Solavetivone 1.643 1.19 NO YES 1.453 0.874 YES (0)

Solafuranone 1.865 1.947 NO YES 2.151 0.557 YES (0)

Scopoletin 1.95 1.378 NO NO 0.516 1.614 YES (0)

N-p-trans-
Coumaroyltyra
2.17 1.271 NO NO 1.008 1.514 YES (0)
mine

Isofraxidin 2.326 1.825 NO NO 0.431 1.862 YES (0)

Lauric acid 1.511 2.89 NO YES 0.954 -0.084 YES (0)

𝛽-sitosterol 2.552 0.855 NO NO 0.43 -1.802 YES (1)

Diosgenin 1.921 1.452 NO NO 0.399 0.247 YES (1)

Acetazolamide 2.292 2.143 NO NO 0.239 2.895 YES (0)

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3.3. Interaction of Standard Drug (Acetazolamide) with 6O4L

Fig. 2. Docking scores and binding interaction of Acetazolamide (PDB ID: 6O4L). The
ligand is shown in line and stick representation along with its 2D diagram and
hydrogen bond interaction.

3.4. Interactions of phytoconstituents with 6O4L A.Solafuranone

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B.Isofraxidin

Fig. 3. Docking scores and binding interaction of phytoconstituents (PDB ID: 6O4L).
The ligand is shown in line and stick representation along with its 2D diagram and
hydrogen bond interaction.

3.5. Boiled Egg

Fig no. 4. Combined Boiled Egg Diagram

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Table no. 4. Name of molecules contained in Boiled Egg Diagram

MOLECULE NUMBER MOLECULE NAME

1 Solanidine
2 Solasodine
3 Solavetivone
4 Solafuranone
5 Scopoletin
6 N-p-trans-Coumaroyltyramine
7 Isofraxidin
8 Lauric acid
9 𝛽-sitosterol
10 Diosgenin
11 Acetazolammide

BOILED means Brain Or IntestinaL EstimateD permeation predictive model. The boiled
egg diagram shows two regions white region and yellow region.

The white region is the physicochemical space of molecules with highest probability of being
absorbed by the gastrointestinal tract, and the yellow region (yolk) is the physicochemical
space of molecules with highest probability to permeate to the brain.
In addition, the points are coloured in blue if predicted as actively effluxed by P-gp (PGP+)
and in red if predicted as non-substrate of P-gp (PGP-).
4. CONCLUSION
In this study, we have carried out an in-silico screening of the phytoconstituents of Solanum
indicum plant. This study demonstrated the sixteen compounds from Solanum indicum plant,
(Solanidine, Solasodine, Solavetivone, Solafuranone, Scopoletin, N-p-trans-
Coumaroyltyramine, Isofraxidin, Lauric acid, 𝛽-sitosterol, Diosgenin). The selected
phytocompounds showed docking scores ranging from –10.0 to −3.7 kcal/mol in 6O4L.
Among all, Solafuranone and Isofraxidin gave the highest binding energy (−7.8 kcal/mol)
and (−6.1 kcal/mol) in complex with 6O4L, whereas the reference compound,
Acetazolamide showed a docking score with a binding energy of -5.1 kcal/mol. Furthermore,
these ligands exhibited good ADMET properties. To summarize, phytoconstituents present
in Solanum indicum possess strong effects against 6O4L and could be further evaluated for
their antiepileptic effect, as well as for the development of alternative drugs with fewer side
effects for the treatment of epilepsy.

REFRENCES
1. Saxena HO, Parihar S, Pawar G, Choubey SK, Dhar P. Phytochemical screening and
HPTLC fingerprinting of different parts of Solanum indicum L.: A dashmool species.

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Journal of Pharmacognosy and Phytochemistry. 2021;10(1):1935-41.

2. Bahgat A, Abdel‐Aziz H, Raafat M, Mahdy A, El‐Khatib AS, Ismail A, Khayyal MT.
Solanum indicum ssp. distichum extract is effective against l‐NAME‐induced
hypertension in rats. Fundamental & clinical pharmacology. 2008 Dec;22(6):693-9.
3. Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, Olson AJ.
AutoDock4 and AutoDockTools4: Automated docking with selective receptor
flexibility. Journal of computational chemistry. 2009 Dec;30(16):2785-91.
4. Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE.
UCSF Chimera— a visualization system for exploratory research and analysis. Journal
of computational chemistry. 2004 Oct;25(13):1605-12.
5. Liles JT, Corkey BK, Notte GT, Budas GR, Lansdon EB, Hinojosa-Kirschenbaum F,
Badal SS, Lee M, Schultz BE, Wise S, Pendem S. ASK1 contributes to fibrosis and
dysfunction in models of kidney disease. The Journal of Clinical Investigation. 2018 Oct
1;128(10):4485-500.
6. Verdonk ML, Cole JC, Hartshorn MJ, Murray CW, Taylor RD. Improved protein–
ligand docking using GOLD. Proteins: Structure, Function, and Bioinformatics. 2003
Sep;52(4):609-23.
7. Leach AR, Shoichet BK, Peishoff CE. Prediction of protein− ligand interactions.
Docking and scoring: successes and gaps. Journal of medicinal chemistry. 2006 Oct
5;49(20):5851-5.
8. SAMANT L, Javle V. Comparative docking analysis of rational drugs against COVID-
19 main protease.
9. Arora S, Lohiya G, Moharir K, Shah S, Yende S. Identification of potential flavonoid
inhibitors of the SARS-CoV-2 main protease 6YNQ: a molecular docking study. Digital
Chinese Medicine. 2020 Dec 1;3(4):239-48.
10. Shah S, Chaple D, Arora S, Yende S, Moharir K, Lohiya G. Exploring the active
constituents of Oroxylum indicum in intervention of novel coronavirus (COVID-19)
based on molecular docking method. Network Modeling Analysis in Health Informatics
and Bioinformatics. 2021 Dec;10:1-2.
11. Kaloni D, Chakraborty D, Tiwari A, Biswas S. In silico studies on the phytochemical
components of Murraya koenigii targeting TNF-α in rheumatoid arthritis. Journal of
Herbal Medicine. 2020 Dec 1;24:100396.
12. Joshi T, Sharma P, Joshi T, Chandra S. In silico screening of anti-inflammatory
compounds from Lichen by targeting cyclooxygenase-2. Journal of Biomolecular
Structure and Dynamics. 2020 Aug 12;38(12):3544-62.
13. Nisha CM, Kumar A, Vimal A, Bai BM, Pal D, Kumar A. Docking and ADMET
prediction of few GSK- 3 inhibitors divulges 6-bromoindirubin-3-oxime as a potential
inhibitor. Journal of Molecular Graphics and modelling. 2016 Apr 1;65:100-7.
14. Tsujimura S, Tanaka Y. Disease control by regulation of P-glycoprotein on lymphocytes
in patients with rheumatoid arthritis. World journal of experimental medicine. 2015 Nov
11;5(4):225.

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33
Deep Discoveries in Chemistry: Unraveling the Mysteries at the Molecular
Level
Dr. Prashant R. Mahalle*
Assistant Prof. and Head, Department of Chemistry
Late B. S. Arts, Prof. N. G. Science & A. G. Commerce College, Sakharkherda
Email: [email protected]
Abstract:
This research article delves into the profound discoveries and advancements in the field of
chemistry, exploring breakthroughs that have unraveled the mysteries at the molecular level.
From the elucidation of complex chemical reactions to the design of novel materials with
unprecedented properties, these deep discoveries have significantly impacted various scientific
disciplines and technological applications. This article provides an overview of key
developments, showcasing the depth and breadth of contemporary research in chemistry.
Key Words: Chemistry Discoveries, Molecular-Level Understanding, Sustainable Practices,
Environmental Chemistry, Bottom-Up Nanofabrication etc.
Introduction:
Deep discoveries in science, especially those at the molecular level in fields such as
chemistry, play a crucial role in advancing scientific knowledge and driving technological
innovation. Deep discoveries provide a fundamental understanding of the underlying principles
governing natural phenomena. This knowledge serves as the basis for developing theories and
frameworks that explain complex processes at a molecular level1. Scientific breakthroughs
often translate into technological innovations. Discoveries at the molecular level can lead to
the development of new materials, processes, and technologies with applications ranging from
medicine and electronics to energy and environmental management. Deep discoveries
stimulate curiosity2 and open up new avenues for research. Discoveries in molecular biology
and pharmacology contribute to the development of new drugs and therapies.
Deep discoveries in chemistry contribute to the development of green and sustainable
practices. Advances at the molecular level often contribute to the convergence of technologies3.
For example, discoveries in nanotechnology, which deals with materials at the nanoscale, have
implications not only in material science but also in medicine, electronics, and energy storage.
Deep discoveries have the potential to revolutionize entire industries. For example,
advancements in molecular electronics may lead to the development of smaller, faster, and
more efficient electronic devices, transforming the semiconductor industry. In summary, deep
discoveries in science, particularly at the molecular level, are instrumental in shaping the
scientific landscape and driving technological progress. They provide the foundation for
innovations that have far-reaching implications across diverse fields, contributing to the
betterment of society and the advancement of human knowledge.
Objectives and Materials and Methods:
Several discoveries in the field of chemistry have challenged traditional understanding, leading
to paradigm shifts and opening new avenues for synthesis. Here are a few notable examples:
1. Molecular Machines and Nanotechnology: The development of molecular machines, such
as synthetic molecular motors and switches, challenges traditional notions of static molecular

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structures. Researchers have created nanoscale machines capable of controlled movement,

enabling potential applications in drug delivery, nanoelectronics, and other fields.
2. Metathesis Reaction: The discovery of metathesis reactions, particularly the development
of olefin metathesis by Grubbs4, Schrock, and Chauvin, challenged traditional thinking about
the limitations of organic synthesis. This groundbreaking discovery provided a powerful tool
for creating carbon-carbon double bonds in a controlled manner, revolutionizing the synthesis
of complex organic molecules.
3. Click Chemistry: Click chemistry, introduced by K. Barry Sharpless and others, represents
a set of highly efficient and selective reactions that challenge traditional synthesis methods.
These reactions, characterized by their reliability and simplicity, have found widespread use in
various applications, including drug discovery and materials science5.
4. Molecular Electronics: The field of molecular electronics challenges traditional
semiconductor technology by exploring the use of individual molecules as electronic
components. Researchers aim to create electronic devices at the molecular scale, potentially
leading to smaller, faster, and more energy-efficient electronics.
5. Green Chemistry Principles: The emergence of green chemistry6 challenged traditional
chemical practices by emphasizing the importance of designing environmentally friendly
processes. This approach considers the environmental impact at every stage of chemical
production, leading to the development of more sustainable and less toxic chemical syntheses.
6. C–H Activation: Traditional organic synthesis often involves functionalizing carbon-
hydrogen (C–H) bonds, which were once considered unreactive. Recent advancements in C–H
activation chemistry challenge this convention, allowing chemists to directly functionalize C–
H bonds, streamlining synthetic routes and reducing the number of steps required for complex
molecule synthesis.
7. Supramolecular Chemistry: Supramolecular chemistry challenges traditional views by
focusing on non-covalent interactions to create complex, hierarchical structures. The field
explores the assembly of molecules into larger, functional structures, leading to applications in
materials science, drug delivery7, and catalysis.
8. Chemoselective Reactions: Discoveries in chemoselective reactions challenge traditional
chemical methods by enabling the selective modification of one functional group in the
presence of others. This has profound implications in the synthesis of complex molecules,
where multiple functional groups may be present.
9. Metal-Organic Frameworks (MOFs): The development of metal-organic frameworks8
challenges traditional notions of solid-state chemistry. MOFs are porous materials with high
surface areas, and their design and synthesis have opened up new possibilities for gas storage,
catalysis, and separation processes.
10. Directed Evolution of Enzymes: The directed evolution of enzymes challenges the
traditional concept that enzymes have fixed structures and functions. Through directed
evolution, researchers can engineer enzymes with enhanced or entirely new functionalities,
leading to applications in pharmaceuticals, biofuels, and more.
These examples illustrate how breakthrough discoveries in chemistry have questioned
established norms, prompting researchers to explore novel avenues for synthesis and
innovation. These paradigm shifts contribute to the continuous evolution and advancement of
the field.

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Conclusion:
From the above points of informative aspects, deep discoveries represent a journey into
the unknown, reshaping our understanding of the molecular world and propelling technological
advancements. They underscore the dynamic nature of science, where each revelation not only
expands our knowledge but also sparks innovation that has far-reaching consequences for
society. As we continue to unravel the mysteries at the molecular level, the transformative
impact on both scientific understanding and technological applications will undoubtedly
persist, driving the evolution of science and technology.

References:
1. Cramer, R. E., & Truhlar, D. G. (2009). A universal approach to solvation modeling.
Accounts of Chemical Research, 42(6), 841-850. DOI: 10.1021/ar800249u.
2. Whitesides, G. M. (2015). Is the focus of most chemistry journals too narrow? ACS
Central Science, 1(1), 16-18. DOI: 10.1021/acscentsci.5b00029.
3. Nørskov, J. K., Bligaard, T., Rossmeisl, J., & Christensen, C. H. (2009). Towards the
computational design of solid catalysts. Nature Chemistry, 1(1), 37-46. DOI:
10.1038/nchem.121.
4. Grubbs, R. H. (2003). Handbook of metathesis, Wiley.
5. Chhowalla, M., Shin, H. S., Eda, G., Li, L. J., Loh, K. P., & Zhang, H. (2013). The
chemistry of two-dimensional layered transition metal dichalcogenide nanosheets. Nature
Chemistry, 5(4), 263-275. DOI: 10.1038/nchem.1589.
6. Anastas, P. T., & Warner, J. C. (1998). Green chemistry: Theory and practice. Oxford
University Press.
7. MacArthur, E. H., & Warren, W. S. (2003). Ultrashort pulse lasers in medicine. Lasers in
Surgery and Medicine, 32(4), 283-305. DOI: 10.1002/lsm.10150.
8. Yaghi, O. M., & Li, H. (1995). Hydrothermal synthesis of a metal-organic framework
containing large rectangular channels. Journal of the American Chemical Society,
117(34), 10401-10402. DOI: 10.1021/ja00141a008.

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34
Synthesis, Characterization of (3-Chloro-benzylidene) – (4-Phenyl-thiazol-
2-yl) amine and Studied their antioxidant Activity
Ghodile R.D1, Chaudhari P.S.2, Dharamkar R. R3.
1
Department of Chemistry, S.P.M. Science & Gilani Arts Commerce College, Ghatanji Dist:- Yavatmal-
445301 India Email: [email protected]
2
Department of Chemistry Vitthal Rukhmini College Savana Ta-Mahagaon Dist- Yavatmal (MS) India
3
Department of Chemistry, G. S. Science, Arts, & Commerce College Khamgaon Dist. Buldhana (MS) India

Abstract -
In the current study, substituted aromatic aldehydes (2a-c) were combined with 2- amino-4-
phenyl thiazole to create a series of 4-phenyl-thiazolyl substituted Schiff bases (3a-c)
derivatives. The IR and 1H-NMR of all the produced compounds validated their structures and
its antibacterial activity was examined. All the title compounds were screened for their
antibacterial activity. Most of the compounds exhibited moderate antimicrobial activity.
Keywords: Schiff’s base; Aromatic Aldehydes; 3-Chloro-benzylidene) – (4-Phenyl-thiazol-2-
yl) amine, Potassium ferricyanide.
Introduction-
In pharmaceutical chemistry, thiazole derivatives have been extremely important. Thiazole
have a wide variety of pharmacological properties, including antimicrobial1-4, analgesic6-7,
anticonvulsant8-9, and antioxidant10, hypolipidemic11, anti HIV-1 12-13, adenosine receptor
antagonist14-15 and osteoporosis inhibitor16. For the creation of antimalarial drugs, Schiff bases
have proven to be intriguing moieties17–18. As potential antibacterial agents, Schiff bases have
been mentioned19–21. It is imperative to find and create more powerful antifungal medicines,
and several Schiff bases are well-known to be effective ones22.
Reaction scheme

Experimental Section
General Conditions: Melting points were discovered using open capillary tubes and are
uncorrected. Iodine spotting and TLC were applied to the silica gel-G surface. Nicolet
5ZDXFT-IR spectrometers in the KBr phase and Brucker WP 200 and 500 SY spectrometers
in the 1H NMR phase were used to record the IR spectra. Preparation of [(3-chloro-
benzylidiene)-4-phenyl-thiazole-2-yl)-amine: general technique (3a) In the presence of HCl,
(0.001mole) 2- amino 4- phenyl thiazole and 0.70gm of 3- chlorobenzaldehyde were refluxed
for three to four hours. The product, M.P.-150-1540C, was obtained from the resultant solid
after it had been filtered, cleaned, and recrystallized from ethanol. 3a: 1HNMR: 2.6 (s,1H,
CH3), 7.2-8.7 (m, 10H, Ar-H), IR (KBr): 1575 cm-1 (C=N)

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IR Data

BRUKER
13.0003

AVANCE II 400 NMR

8.9334
8.3077
8.1304
7.8549
7.8519
7.8336
7.5923
7.5885
7.5847
7.5696
7.5673
7.4795
7.4604
7.4398
7.4026
7.3838
7.3388
7.3212
7.2490
7.1560
7.1510
7.1397
7.1203
7.0957
7.0166
4.1678
4.1575
3.1373
3.1197
3.1013
2.5378
2.5127
2.5088
2.5054
1.2804
1.2621
1.2438
1.1826
Spectrometer
SAIF
Panjab University
Chandigarh

Current Data Parameters

NAME Mar14-2018
D EXPNO 180
PROCNO 1

F2 - Acquisition Parameters
Date_ 20180314
Time 18.26
INSTRUM spect
PROBHD 5 mm PABBO BB-
PULPROG zg30
TD 65536
SOLVENT DMSO
NS 8
DS 2
SWH 12019.230 Hz
FIDRES 0.183399 Hz
AQ 2.7263477 sec
RG 144
DW 41.600 usec
DE 6.00 usec
TE 299.3 K
D1 1.00000000 sec
TD0 1

======== CHANNEL f1 ========

NUC1 1H
P1 10.90 usec
PL1 -3.00 dB
SFO1 400.1324710 MHz

F2 - Processing parameters
SI 32768
SF 400.1300000 MHz
WDW EM
SSB 0
LB 0.30 Hz
GB 0
PC 1.00
1.00

0.87

0.39
3.09
1.66
2.06

15 14 13 12 11 10 9 8 7 6 5 4 3 2 1 0 ppm

[email protected]
Sr. No. Frequency in cm-1 Assignment Literature value cm -1

1 3194.26 Ar -H 3300-2900
2 1608.70 C=N stretching 1640-1690
3 1473.68 C=C stretching 1400-1500

Antioxidant
Ferric reducing antioxidant power assay
Reducing power assay method for determination of antioxidant activity/ potential of
samples
Procedure:-

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Different concentration of the drug (10-50ug/ml) was added to 2.5 ml of 0.2M sodium
phosphate buffer (pH 6.6) & @.5ml of 1% potassium ferricynide [K3Fe(CN)6] solution. The
reaction mixture was vortexed well & then incubated at 500 C for 20 min using vortex shaker.
At the end of the incubation , 2.5 ml of 10% trichloacetic acid was added to the mixture and
centrifuged at 3,000 rpm for 10min. The supermantant (2.5ml) was mixed with 2.5 ml of
deionized water & 0.5ml of 0.1% ferric chloride. The coloured solution was red at 700nm
against the blank with reference to standard using UV Spectrophotometer. Here, ascorbic acid
was used as a reference standard.
The percentage of radical scavenging (%) was calculated by the following formula:
% Free radical scavenging activity = Ac – As /Ac × 100
Where , Ac = Absorbance of control at 720 nm
As = Absorbance of sample
The concentration of sample required to scavenge 50% of the DPPH free
radical (IC50) was determined from the curve of percent inhibitions
plotted against the respective concentration.
Experimental
Absorbance should increses with increses in concentration. Results confirm the reducing power
Of samples which increased with increasing concentration. Higher value absorbance of the
reaction mixture indicated greater reduing power. The redusing power was found to be on
oreder Br>NO2.
Table 1 Ferric Reducing Antioxidant Power Assay
Standard Observation - (Ascorbic acid) (720nm)
Concentration(ug/ml) Absorbance 1 Absorbance 2 Absorbance 3
10ug/ml 0.002 0.006 0.061
20ug/ml 0.395 0.251 0.029
30ug/ml 0.941 0.775 0.499
40ug/ml 1.451 1.350 1.036
50ug/ml 2.000 2.000 2.000
Control Absorbance - 0.812 (720nm)
Sample Observation- (720nm)
Formula –
Scavenged(%) = (Acont) - (Atest) /(Acont) ×100
Table2 (% Scavenged of DPPH free radical)
Sample name Concentration Absorbance %Scavenged
4a (H) a) 30ug/ml 0.121 72.1 %
b) 50ug/ml
0.305
4b (OCH3 ) c) 30ug/ml 0.213 68.2 %
d) 50ug/ml
0.504
4c (NO2) a) 30ug/ml 0.068 62.9%
50ug/ml
0.463

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4d (Br) e) 30ug/ml 0.171 17.1%

f) 50ug/ml
0.629
4e (Cl) a) 30ug/ml 0.058 21.2%
b) 50ug/ml
0.102

Results and Discussion

Several five member aromatic systems having three hetero atoms at symmetrical position
1,3,4-thiadiazole had been studied because of their interesting physiological properties.
Specifically substituted benzaldehyde or naphthaldehyde were combined with 2-amino-5-
diethylamino-pentane in an equal amount of methanol to produce the desired Schiff-base
phenols or Schiff base naphthalene ligands for evaluation 19. Furthermore, by reducing their
corresponding Schiff-base counterparts with KBH4 at 80°C for 3 hours, substituted amine
phenol or amine naphthalene ligands were also produced 20.
All compounds underwent standard analytical procedure purification and characterization.
The spectral and analytical data for digits 1 through 7 agreed with the suggested formulation.
By measuring the incorporation of 3 H hypoxanthine, we assessed 1-7's capacity to prevent the
formation of trophozoites in intraerythrocytic culture [21]. A more accurate way to quantify
parasite development is through the inhibition of hypoxanthine incorporation, which correlates
well with direct blood smear counts. Unfortunately, neither the CQ-sensitive (HB3) nor the
CQ-resistant (Dd2) strains were significantly affected by compounds 1, 2, 3, 4, and 6. The
moderate half-maximal inhibitory concentration (IC50) values for the active compounds 5 and
7 against CQ-sensitive lines, respectively, were 5.53 and 6.38 M. However, compound 7 was
found to be twice as effective as compound 5 against CQ-resistant Dd2 lines (IC50, 3.5 M for
compound 5; 1.7 M for compound 7).
The structural differences of compounds 5 and 7 may be responsible for their varied
biological actions. The quinoline ring of the CQ, which lacks chlorine at position 7 and the
quinonyl basic nitrogen, has a simple substituted aromatic ring in position 5, but the quinoline
ring of the 7 contains a naphthalene ring. Furthermore, it has been demonstrated previously
that nitrogen atoms of side chain in chloroquine and aromatic ring assist in accumulation of the
drug within the digestive vacuole of the parasite (Vydac) employing an eluent mixture of
methanol and water (100% water for first 5 minutes, followed by a change to 50 : 50 (Water :
MeOH) for the next 5-20 minutes, and finally 100% MeOH for the next 20-25 minutes). The
retention times (Rt) are given for selected compounds.
Absorbance should increses with increses in concentration. Results confirm the reducing power
Of samples which increased with increasing concentration. Higher value absorbance of the
reaction mixture indicated greater reduing power. The redusing power was found to be on
oreder Br>NO2.
Acknowledgment:
The Authors are very thankful to the Department of Chemistry S. P. M. Science and
Gilani Arts, Commerce College Ghatanji and Shri Shivaji Science College Amravati for
provide the necessary facilities in the laboratory. Words of gratitude are also expressed for
SAIF/RSIC Chandigarh for IR and NMR spectral data.

References:
1. P M Bonilla, A P Cardena, E Q Marmol, J L Tellez & G L M Rejon, Heteroatom. Chem., 17(2006)254.
2. K A Parmar, B G Suthar & S. Parajapati, Bull. Korean. Chem. Soc, 31 (2010) 793.
3. N C Desai, N B Bhatt, H C Somani, & A R Trivedi, Eur. J. Med. Chem., 67 (2013).
4. A Idhayadhulla, R S Kumar, A J A Naseer, J. Mexican. Chem. Soc., 55 (2011) 218.

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National Conference on "Recent Advancements in Science & Technology" ISBN : 978-81-19931-25-5

5. A K Prajapati & V P Modi, J. Chil. Chem. Soc., 55 (2010) 240.

6. G Saravanan, V Alagarasamy & C R Prakash, Asian J Res. Pharm. Sci. 1 (2011) 134.
7. N Siddiqui & W Ahsan, Eur. J. Med. Chem., 45 (2010) 1536.
8. N Siddiqui & W Ahsan, Med. Chem.Res., 20 (2011) 261.
9. A Geronikaki, D H Litina, C. Chatziopoulos &G. Soloupis, Molecules, 8 (2003), 472.
10. S N Mokale, P T Sanap & D B Shinde, Eur. J. Med. Chem., 45(2010) 3096.
11. P Vicini, A Geronikaki, M Incerti &B. Busonera, Bioorg. Med. Chem., 11 (2003)4785.
12. W Fathalla, Arkivoc, 12 (2008)245.
13. P Bhattacharya, J T Leonard &K. Roy, Bioorg. Med. Chem., 13 (2005) 1159.
14. A B Scheiff, S G Yerande &El Tayeb, Bioorg. Med. Chem.,18 (2010) 2195.
15. ChSanjeeva Reddy*a , M VaniDevia , M Sunithaa , B Kalyania& A Nagarajb . Synthesis and antibacterial
activity of diheteryl substituted[1,2,4]triazolo[3,4-b][1,3,4]thiadiazoles. Indian journal of chem.vol. 55B
May 2016 pp. 590-597.
16. PramillahSah* &VasudhaKaul.Synthesis and in-vitro biological evaluation of some quinazolin of chem.
Vol. 49B Oct 2010 pp. 1406-1412.
17. Kanti Sharma* & RenukaJain.Synthesis,reaction and anthelmintic activity of 1-[benzimidazol-2-yl]-4-
formyl-3-[2’(- substituted phenyl) indole-3-yl]pyrazoles. Indian journal of chem. Vol. 51B Oct 2012 pp.
1462-1469.
18. K. Taguchi, F. H. Westheimer, J Org Chem, Volume 36, Issue 11, 1971, pp. 1570-1572
19. Wellems, T. Science, 2002, 298, 124.
20. Sharma, V. Mini Rev. Med. Chem., 2005, 5, 337.
21. Ocheskey, J.; Harpstrite, S.; Oksman, A.; Goldberg, D.; Sharma, V. Chem. Commun., 2005, 1622

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35
Synthesis and Characterization of Schiff Bases Using Different Amines and
their Biological Activities
Priyadarshani N Deshmukh
SSES Amt’s Science College,Nagpur
[email protected]

Nowadays, the research field dealing with Schiff base chemistry has expanded enormously.
The importance of Schiff base for bioinorganic chemistry, biomedical applications,
supramolecular chemistry, catalysis and material science, separation and encapsulation
processes, and formation of compounds with unusual properties and structures has been well
recognized and reviewed. They also serve as a back bone for the synthesis of various
heterocyclic compounds. Schiff bases have been utilized as synthons in the preparation of a
number of industrial and biologically active compounds like formazans, 4-thiazolidinines,
benzoxazines, and so forth, via ring closure, cycloaddition, and replacement reactions. Schiff
base are organic compounds possessing azomethine group which resulted from condensation
of amine with aldehyde or ketone. Schiff base derived from aromatic amine and aromatic
aldehydes have a wide variety of application such as biological activity, catalytic activity and
also used as ligands to obtain metal complexes because of their excellent abilities of this type
of Schiff base are widely used as anticorrosion for different metals in different media. The
synthesis and characterization of transition metal complexes of schiff bases containing nitrogen
and oxygen donors atoms has increased manifold in the recent past .The Schiff base ligands
are considered to be good chelating agents [3], Schiff bases are a special class of ligands with
a variety of donor atoms exhibiting interesting coordination modes towards transition metals,
and azomethine linkage is responsible for the biological activities. The Schiff bases derived
from various amines have been widely investigated and find applications in biomimetic
catalytic reactions, materials chemistry and industry. Schiff base complexes have also gained
attention as stereo chemical models in transition metal coordination chemistry due to their
structural variety.
Key Word:- Synthesis, Schiff Bases, Biological Activity.
MATERIALS AND METHOD
Preparation of Schiff Base by using different substituted aromatic amines with substituted
aromatic benzaldehyde. Following materials and method is included.

MATERIALS

Materials Required are -

Substituted Aromatic Amines such as Aniline, p-chloro Aniline,

ANILINE-Aniline appears as a yellowish to brownish oily liquid with a musty fishy

odor.melting point -6℃; boiling point 184℃;flash point 158℉. Denser than
water(8.5lb\gal)and slightly soluble in water. Vapours heavier than air. Toxic by skin
absorption and inhalation.Produces toxic oxides of nitrogen during combustion. Used
tomanufacture other chemicals, especially dyes, photographic chemicals ,agricultural
chemicals and others.

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P-CHLORO ANILINE-4-Chloro aniline is a chloroaniline in which the chloro atom is para

tothe aniline amine group.It is member of monochlorobenzenes. P-chloroaniline appears as a
white or pale yellow solid.Melting Point is 69.5℃.

Substituted Aromatic Benzaldehydesuch as 4-chloro Benzaldehyde

4-CHLOROBENZALDEHYD

-4-Chloro Benzaldehyde appears as colourless to yellow powder or white crystalline solid.It

has pungent odour.
GENERAL METHOD FOR PREPARATION OF SCHIFF BASE-
The mixture of different substituted aromatic amines and substituted aromatic benzaldehyde
was dissolved in methanol . These solution was acidified and reflux for about 5-6 hrs. The
excess of solvent was removed under pressure. The resulting compound was washed, dried and
crystallized.

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ANTIMICROBIAL ACTIVITY OF SCHIFF BASE

The synthesized schiff bases were screened in vitro for their antibacterial activities against
Escherichia coli (E.coli) and Bacillus subtilis ( B. subtilis) bacterial strains using well diffusion
method. The bacterial cultures were incubated at 30± 0.1℃ for 24 hours by inoculation into
nutrient agar.Schiff bases were stored dry at room temperature and 3ml ethanol. Antibacterial
activities of each compound were evaluated by the agar disc diffusion method. Muller hinton
agar media (15 cm3) kept at 45℃ was poured in the petri dishes and allowed to solidify poured
petri plates were streaked with culture media wells formed prepared schiff bases (100μl) were
applied on the solid ager media by pressing tightly. The petri plates were placed at 37℃ for 24
hours.At the end of period the inhibition zones formed on media were measured with zone
reader millimeters.

SAMPLE Conc. (𝛍g/ml) GRAM POSITIVE E. coli GRAM NEGATIVE B.

subtilis

N-(4-CHLOROBENZYLIDENE 100𝛍L 10mm 12mm

ANILINE)
(1C)
100 μl - 13mm

N-(4-CHLORO BENZYLIDENE)-4- 100𝛍l 10mm 16mm

CHLORO ANILINE
(2C)
100 μl - 14mm

Antibacterial activities against Schiff compound at concentration 100𝛍l/ml.

REFERENCES
1. Pouralimardan, O; Chamayou, A. C; Janiak, C; Monfared, H. H, Inorg. Chim.Act. 360,2007, 1599.
2. Krishnapriya, K. R.; Kandaswamy, M.; Polyhedron, 2005, 24, 113.
3. ooysena, I. N.; Maikooa, S.; Akermana, M. P.; Xulua, B.; Munro, O. J. Coord.Chem., 2013, 66(20),
3673.
4. Chandra, S; Jain, D; Sharma, A. K; Sharma, P, Molecules 2009, 14, 174.
5. Sinha, D.; Tiwari, A. K; Singh, S; Shukla, G; Mishra, P; Chandra, H.; Mishra, A. K. Eur. J. Med.
Chem. 2008, 43, 160.
6. Ansary, E.; Soliman, A. L.; Sherif, A.A.; Ezzat, J. A. Synth. React.
Inorg.MetOrg.Chem.,2002,32(7).1301.
7. Celik,C.;Tumer,M.;Serin,S.Synth.React.Inorg. Met-Org.Chem.,2002,32(10)1839.
8. Biswas,C.;Drew,M.G.B.;Figuerola,V.;GomezCoca,S.;Ruiz,E.;Tangoulis,V.;Ghos h,A.;
Inorg.Chim.Act.,2010,363,846.
9. Schiff,H.Justus Liebigs Ann. Chem.1864,131,118.
10. R.HernandezMolina,A.Medros,ComprehensiveCoordinationChemistry II,2003,411-446.
11. Patai S. The Chemistry of the carbonnitrogen double bond, John Wiley & Sons ltd., London,1970.
12. Arora K. and Harma K. P. Synth. React. Inorg. Met.-Org. Chem. 2003, 32, 913.
13. Nimitsirwat, N.; Vernon, C. J Am Chem Soc 2004, 32,126.
14. Mirkin, M.V. and Bard, A.J. J. Anal. Chem. 1991, 63, 532.
15. Kratz, F.; Beyer, U.; Schutte, M. T. Crit. Rev. Ther. Drug 1999, 16, 245.
16. Saito, H.; Hoffman, A. S.; Ogawa, H. I. J. Bioact. Compat. Polym. 2007, 22, 589
17. Dhar DN, Taploo CL. J Sci Ind Res. 1982, 41, 501.
18. Przybylski P, Huczynski A, Pyta K, Bartl, B. Curr. Org. Chem. 2009, 13,124.

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36
Physico-Chemical Assessment of Soil Sample in Jarud Region in Amravati
District of Maharashtra (India)
1Dr. Priyanka U. Belsare and 2Ku Tejashri M. Ghotkar.
1
Department of Chemistry, Mahatma Fule Arts, Commerce and Sitaramji Chaudhari Science Mahavidyalaya,
Warud. Dist. Amravati-444906 (M.S.) India.
2
Department of Chemistry, Art’s and Commerce College, Jarud. Dist. Amravati (M.S.) India.
Corresponding author: [email protected]

ABSTRACT
Soil is a natural body of mineral and organic material differentiated into horizons,
which differ among themselves as well as from underlying materials in their morphology,
physical make-up, chemical composition and biological characteristics. In the present research
work, studies on soils with physical properties, chemical properties and micronutrients of soils
have been done. Soil samples were collected from two different locations covering Jarud, in
Warud Tahsil in Amravati District (Maharashtra) India. The soil parameters like soil colour,
moisture, pH, Carbon, Nitrogen, Copper, Potassium, Phosphorous, Zinc, Boron, Copper and
Sulphur content, were analyzed in the month of March 2022. The values of pH indicated that
all samples of the soils are alkaline, all samples were containing moderate amount of available
micronutrients.
Keywords: Physico-Chemical analysis, Parameters, Jarud Village, Plants Nutrients.

INTRODUCTION
To improve soil, plant health and soil balance to optimize production, it’s important to
identify the soil’s biological, mineral, chemical and physical requirements. When evaluating
the soil analysis, we look for synergies and approach mineral and microbe plant and pest
management holistically to provide the formula for nutrient, trace elements and biological
inputs to achieve true soil balance. The soil analysis report provides the information necessary
to set nutrient application targets, which are used to calculate manure and fertilizer application
rates. Test results from regular field sampling (particularly from benchmark sites) allow
monitoring and detection of changes in soil parameters (e.g., nutrients, pH, and salinity) with
time.
Soil analysis provides us the background knowledge on soil available nutrient status
along with some physico-chemical and biological properties; that we need in order to take
specific decisions on balanced and integrated nutrient management for healthy soils and crops.
The quality and health of soil determine agricultural sustainability and environmental quality
which jointly determines plant, animal and human health. The status of available
micronutrients in soils and their relationship with various physico-chemical properties have
been attempted by several investigators (1-2). Considerable research work has been done
regarding the study of Nutrients and Physico-Chemical assessment of various types of soil in
Maharashtra as well as in India (3-4) but the investigation of nutrients and parameters of soil
of Jarud village in Amravati district in Maharashtra.

Physico-chemical characteristics of some samples of soil from some farms of nearby

villages of Kannad taluka, Dist. Aurangabad (5). Abhishek Jangir et. al. (6) evaluates the major
and micronutrient status of Kelapur block, Yavatmal district, Maharashtra. Wodaje Addis et.
al. (7) carried out assessment of some selected physicochemical parameters in soil samples
collected from four agricultural areas of East Gojjam Zone (Debrem Markos, Dejen, Bichena

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and Debre (Werk), Ethiopia. Perveen et. al. (8) has studied micronutrient status of soils and
their relationship with various physico-chemical properties. Khadke et. al. (9) reported soil
analysis and its environmental impact on Nanded city of Maharashtra State. The impacts of
industrial pollution on the ground water soil and plant have also been reported in our country
and abroad (10). Investigation of some parameter and Nutrients from Soil samples of Rice field
by Jadhav et.al. (11). Geeta Tewari et. al. (12) carried out the physico-chemical properties of
soils from different land use systems viz. agriculture, olericulture and two dominant forest
types.
R. Zornoza et al (13) studied the Soil quality (SQ) assessment has long been a
challenging issue, since soils present high variability in properties and functions. K.
Senthilarasan et. al. (14) worked on soil analysis to study physical properties and
micronutrients of soils. Soil samples were collected from five different locations covering
Nagapattinam district (south) in tamilnadu. Joshi et al. concluded that India Soil salinity greatly
affects growth and development of vegetation (15), but the investigation of nutrients and
parameters of Soil of Jarud village in Warud Tahsil of Amravati district in Maharashtra, India
was still lacking.

EXPERIMENTAL
Study Area
Mauja Jarud (part2) is a small village in Warud Tahsil of Amravati District
(Maharashtra State). This area is well known for orange, cotton and Toor. The sources of water
for this area are of well. Relative to its geographical location, the study area enjoys a tropical
type of climate.
Collection of Samples
Soil samples were collected randomly at 0 to 15 cm and 15 to 30 cm depths with five
plots (field), five samples from each plot (field) respectively, in well sterilized polythene
pouches. Soil sample were collected from following Farmers fields.

1) Sample 1 (TMG-1) was collected from Mr. Manikrao Sitaramji Ghotkar field.
2) Sample 2 (TMG-2) was collected from Mr. Raju Devrao dathe field.
Table-1: Nutrients and Physico-Chemical assessment of Soil Samples.

Analysis of Parameters
Sr.No. Parameters TMG-1 TMG-2
1 Colour Black Whitish Black
2 pH 7.41 7.42
3 Organic Carbon (%) 0.46 0.45
4 Nitrogen 210 200
5 Phosphorous (kg/hect.) 28 31
6 Potassium (kg/hect.) 340 350
7 Mn (ml/gm/kg) 8 7
8 Fe (ml/gm/kg) 39 39
9 Zinc (ml/gm/kg) 3.1 3.1
10 Boron 1.5 1.5
11 Copper 8 9
12 Sulphur 59 59
13 Salinity 0.2 0.2

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Determination of Physical Parameters

Reagents uses for this research work were AR grade and chemicals other than reagent
are LR grade manufactured by S.D. fine, LOBA and Merck. The soil samples were dried for
about 24 hr. and grinded more finely. In this work about thirteen physico-chemical parameters
and nutrients of two soil samples were determined and results were also recorded. During
collection, temperature of the sample was recorded.

Following methods were used for estimation of various parameters are:

i. Determination of Moisture: by Weighting Method.

ii. Determination of pH: by Digital pH Meter
iii. Determination of Organic Carbon: by Titration Method
iv. Determination of Copper (Cu): by Atomic Adsorption Spectroscopy.
v. Determination of Nitrogen (N): by Titration Method
vi. Determination of Phosphorous (P): by Titration Method
vii. Determination of Potassium (K): by Flame Photometry
viii. Determination of Color of Soil: by Viewing soil

RESULTS AND DISCUSSION

Colour
The soil sample TMG-1 was black in colour and sample TMG-2 was whitish black in colour.
Moisture
Moisture content value ranges from 1% - 4%. It is clear from the result that Soil sample TMG-
1and TMG-2 has 2.5 and 3.7% moisture respectively.
pH
pH was medium quantity in the ranges 7.41 to 7.42. To dig drains, to make out flows, sowing
green crops, cow manure to burry plant wastes, to supply necessary amount of water, use of
gypsum is need.
Organic carbon
Organic carbon contents were recorded in the range 0.46-0.45 %. The soil sample TMG-1
and TMG-2 has medium organic carbon.
Nitrogen
Available Nitrogen quantity ranged from 200– 210 Kg/hect. The soil sample TMG-1 and
TMG-2 has less Nitrogen content.
Phosphorous
Available Phosphorous was medium quantity ranged from 28-31kg/hect. The soil sample
TMG-1 have less Phosphorous as compared to TMG-2.
Potassium
Available Potassium quantity ranged from 340-350 kg/hect. The soil sample observed value
for potassium was medium.
Zinc
The Zinc was less in quantity in both TMG-1 and TMG-2 i.e 3.1 ml/kg.
Copper
Copper content is soil sample ranges from 8-9. It has been seen that in soil sample TMG-1
and TMG-2 has approximate quantity of copper.
Boron
It has been seen that in soil sample TMG-1 and TMG-2 has 1.5 ml/gm/kg of Boron. Both the
soil sample contains approximate Boron quantity.
Sulphur

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It has been seen that in soil sample TMG-1 and TMG-2 has 59 ml/gm/kg of Sulphur. Both the
soil sample contains approximate Sulphur quantity.

CONCLUSION
In the soil sampleTMG-1and TMG-2, the pH becomes middle alkali and to increases
the pH of this soil suggested the use of compost manure. Then recommendation of to dig drains,
to make out flows, sowing green crops, cow manure to bury plant wastes, to supply necessary
amount of water, use of gypsum. In the soil sampleTMG-1 and TMG-2, Zinc becomes less,
then recommendation zinc sulphate10 kg / hectare. In the soil sample TMG-1 and TMG-2 the
potassium become medioum, then recommendation Potash 400gm / hectare. In the soil sample
TMG-1 and TMG-2 the phosphorous is middle. To increase the crop quality phosphorous will
be given through fertilizer 2000 gm /hectare. In the soil sampleTMG-1 and TMG-2, Boron
becomes approximate. Organic fertilizer from goat manure resulted in the highest yield per
hectar in groundnut and total biomass in palm oil plant. In the soil sampleTMG-1 and TMG-2
sulphur become approximate. In the soil sample TMG-1 and TMG-2 medium organic carbon
and nitrogen. So, to increase more yield of crop by using carbon and nitrogen to soil in the
form of compost manure.
ACKNOWLEDGEMENT
The authors are thankful to Dr. G. N. Chaudhari, Principal, Mahatma Fule Art’s, Commerce
and Sitaramji Chaudhari Science Mahavidyalaya, Warud, Dist. Amravati and Non-teaching
staff for providing necessary laboratory facility during this work. Also thankful to Soil testing
Lab, Kisan foundation Warud, Amravati for their valuable suggestions.

REFERENCES
1. M. Kumar and A. L. Babel, Indian Journal of Agricultural Science, 3, 97 (2011).
2. Rajesh P. Ganorkar, Harshali A. Hole and Dinesh A. Pund, Rasayan J. Chem., 10(2), 429-
433(2017)
3. R. P. Ganorkar and P. G. Chinchmalatpure, Int. J. Chemical, Env. And Pharmaceutical
Research, 4(2&3), 46(2013).
4. R. P. Ganorkar and N.H. Khan, International Journal of Chemical and Pharmaceutical
Analysis, 1(4), 190(2014).
5. Dhananjay Vyankat Bondar Physico-chemical analysis of soil (2022).
6. Abhishek Jangir, R.P. Sharma, G. Tiwari, D. Vasu1, S. Chattaraj, B. Dash1, L.C. Malav,
P. Chandran, S.K. Singh, H. Kuchankar and S. Sheikh issn: 022-457x ,241-245, (2019)
7. Wodaje Addis and Alemayehu Abebaw ISSN: 2226-7522 Science, Technology and Arts
Research Journal Sci Technol. Arts Res. J., (2014)
8. S. Perveen, M. Tariq, J. K. Farmanullah and A. Hamid, Journal of Agriculture, 9(5), 467,
(1993).
9. Khadke P. A., Bhosle A.B. and Yennawar V. B, Research Front,1(1) (2013)73.
10. Maliwal, G.L.et al, Poll. Res., 23(1), (2004)169.
11. Jadhav S.D, Sawant R.S. and Godghate A.G., Res. J. Agriculture and Forestry Sci., 1(4)
(2013)24.
12. Geeta Tewari, Deepti Khati, Lata Rana, Poonam Yadav, Chitra Pande, Sunita Bhatt,
Vinod Kumar, Neeta Joshi and Prasoon K. Joshi, J. Chem. Eng. Chem. Res. Vol. 3, No.
11, 2016, pp. 1114-1118, (2016)
13. R. Zornoza et al. SOIL, Vol. 1, 173–185, (2015).
14. K. Senthilarasan, P. Sakthivel, A. Jenifer1, T. Susmitha, G. Janakiaman, Volume 5, Issue
4, ISSN-2349-5162 (2018).
15. V. D. Joshi, N. P. Narhari, & P. R. Rachh, Int. Jr. of chemical Reserch, 1(3), 709 – 713,
2009.

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37
Synthesis ,Characterization And Antimicrobial Study Of Co(II),Cu(II) and
Fe(III) Complexes Of Substituted 4,4'- Dimethoxybenzoinhydrazones
P.M.Dahikar
Department of Chemistry
J.D.Patil Sangludkar Mahavidyalaya Daryapur, Dist.- Amravati E-mail: [email protected]

Abstract: Metal benzoinhydrazone complexes have been synthesized from substituted

benzoinhydrazone were carried out by the known literature method.The structure of all the
synthesized compounds were justified on the basis of chemical characterized by elemental and
spetral analysis. The Physico-Chemical data suggest octahedral geometry for Co(II) & Cu(II)
complexes. The synthesized complexes were screened for antimicrobial activity at a
concentration of 1000µgm/ml.Which was serially diluted to determine their MIC values.
Keywords:- Metal complexe, 4,4'-Dimethoxybenzoinhydrazones, sodiumhydroxide DMF-
Water(80%) medium, Antimicrobial Activity , spectral analysis.
Introduction:-
The complex formation of benzoinhydrazone with copper(II) and nickel(II) , were synthesized
by scherbakov1 .Prasad2 , studied by the synthesis of Novel(II) complexes with now ligand
derived from hydrazone of isoniazid and their magnetic-spectral, electrochemical, thermal and
antimicrobial investigation. Benzoinhydrazone are well known for their biological activity
coordination compounds containing ONS as donor atoms are reported to antimicrobial
activity3. Synthesis, spectral and biological studies of Co(II), Ni(II), Zn(II), Cu(II) and Cd(II)
complex with benzyl salicyladehyde acyldihydrazone were carried out by singh4. The
simultaneous spectroscopic determination of palladium and osmium with salicyladehyde
hydrazone was carried out by Ray5. The synthesis and structural characterization of three new
co-ordination complexes of Co(II), Mn(II) and Cu(II) with N,N,O- donor hydrazine ligands
were carried out by shit6 .The coordination chemistry of hydrazones is an intensive area of
study and numerous transition metal complexes of these ligands have been investigated7.
Synthesis and characterization of some copper(II) complexes with N,S,O-donor
thiohydrazones were carried out by Dey8. The important reactions of carbonyl with
hydroxylamine, semicarbazide and various hydrazines were briefly studied in presence of
strong base in ethanol medium.9-14.While furoinbenzoinoximes, furoinbenzoin hydrazone,
furoinphenyl hydrazone and furoinbenzoin semicarbazone were synthesized by the interaction
of furoinbenzoin with hydroxylamine hydrochloride, hydrazine hydrate, phenyl hydrazine and
semicarbazide hydrochloride in presence of aqueous sodium hydroxide in DMF-Dioxane-
water (80%) medium respectively.
Experimental:-
The 4,4'-Dimethoxybenzoinhydrazones was prepared by refluxing substituted benzoine with
hydrazine hydrate in presence of alkaline medium for 3-4 hours this reaction mixture was kept
overnight. This solid products formed were isolated and washed several times with water
alcohol mixture the purity was checked by TLC paper. Their structural details were confirmed
on the basis of elemental and spectral analysis.In order to Synthesize the complexes the
equimolar mixture of each of the ligand(0.01M) and metal salts(0.01M) were refluxed on a
water bath for 6,8 hours in presence of sodium acetate in ethanol . The reaction mixtures was
kept overnight. The product formed were isolated washed several times with cold water ethanol

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mixture. The characterization of synthesized complexes was made with elemental analysis, IR
and UV-VIS spectra.

Result and Discussion

TABLE-1
THE METAL COMPLEXES COMPOUND,MOLECULAR WEIGHT, COLOUR AND
ELEMENTAL ANALYSIS OF VARIOUS METAL IONS

Complexes Colour Molecular Elemental analysis Found/(calculated)%

wt
C H N M
4,4'-DMBH-Co(II) Pale 682.93 55.30 4.93 8.19 7.71
(56.22) (5.85) (8.19) (8.62)
4,4'-DMBH-Cu(III) Brown 687.54 54.90 4.87 8.14 8.30
(55.84) (5.81) (8.14) (9.24)
4,4'-DMBH-Fe(III) Brown 627.84 60.24 4.86 8.91 7.96
(61.16) (5.73) (8.91) (8.89)

TABLE-2
IR SPECTRAL DATA OF LIGANDS AND ITS METAL COMPLEXES

Ligands and its ʋ(O-H) ʋ(C=N) ʋ(C-O) ʋ(M-O) ʋ(M-N)

Complexes

4,4'-DMBH 3388 1634 1362 - -

4,4'-DMBH-Co(II) 3351 1614 1308 466 578

4,4'-DMBH-Cu(II) 3327 1618 1317 520 585

4,4'-DMBH-Fe(III) 3378 1610 1249 471 565

The IR spectra of lignad shows a strong band at 1634 cm-1 due to (C=N) group broad band
around 3388 cm-1 in the spectra of complexes is assignable of water.4,4'-DMBH-Co(II) shows
band at 3388 (O-H).Which decreases 3351cm-1 inducting that attached to oxygen .However
1634 (C=N) significantly decrease to 1614 cm-1 showing linkage through azido nitrogen.The
ʋ(M-O)& ʋ(M-N) vibration are verified to existing by the appearance of new weak bands in
the spectra of complexes at 578 & 565 respectively.

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TABLE-3
ELECTRONIC SPECTRAL DATA,MAGNETIC MOMENT AND LIGAND
FIELD PARAMETER OF THE METAL COMPLEXES

Complexes μeff λmax (cm-1) Dq B CFSE % (ɅM)Ω-

1
(B.M.) Cova Cm2
Mol-1

4,4'-DMBH-Co(II) 4.72 13604,19018, 1492 0.737 205 26.03 9.5

23023

4,4'-DMBH-Cu(II) 1.79 13698,18587, 1492 0.737 205 26.29 10.8

22984

4,4'-DMBH-Fe(III) 5.67 13888,17979, 1514 0.646 346 35.04 11.4

22471

The electronic spectrum of Co(II) complexes exhibits three bands at 13604,19018 and 23023
-1 4 4
cm which may be assigned to A2g---› T2g(F) ,4A2g--›4T1g (F) and 4A2g--›4T1g(P) ,transition,
respectively for an octahedral stereochemistry11 .The magnetic moment value of 4.72 B.M for
Co(II) complex is consistent with octahedral geometry around metal centre. Fe(III) Complexes
three bands are observed in case of Fe(III) complexes at 13888,17979,22471 cm-1 which may
6 4
be assigned to A1g---› T1g(F),6A1g--›4T2g(F) and 6A1g--›4Eg, belongs to transition respectively,
indicating octahedral geometry of Fe(III)complexes 12-13. The value of 5.67 B.M. would
suggest high spin six coordination for Fe(III) complexes. The values of various ligand field
parameters 10Dq ,B ,λmax, CFSE and conductance respectively favoring in octahedral
geometry for this complex.

ANTIMICROBIAL ACTIVITY OF COMPLEXES

The compounds were assayed for their antimicrobial activities14 against four test organisms
E.coli, S. aureus, Ps.aeruginosa, B. subtilis at a concentration of 1000 μgm/ml by agar well
technique15. Further their MIC value against these organisms were determined by serial dilution
method using DMF as a solvent. The results obtained are given in table-5
TABLE-5
MIC VALUES IN μgm/ml OF COMPOUNDS

Complexes E.coli S.aureus P. aeruginosa B.Subtilis

4,4'-DMBH-Co(II) 125 250 125 125

4,4'-DMBH-Cu(III) 125 63 63 63

4,4'-DMBH-Fe(III) 125 250 125 125

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On the basis of MIC values, 4,4'-DMBH-Cu(II) is found to be most effective antimicrobial agent
followed by 4,4'-DMBH-Co(II) and 4,4'-DMBH-Fe(III).The enhance antimicrobial activity in
case of the compounds. 44’DMBH-Cu(II) showed the lowest MIC values(i.e 63µgm/ml) against
maximum number of microorganisms.

Acknowledgement
The authors are thankful to the SAIF,CDRI, Lukhnow for IR SAIF Chandigarh .Department of
Physics, SGBAU, Amravati for magnetic measurements of the compounds respectively.

References
1.I.N.Shcherba,I.D.Popov,S.I.LEVCHEKOV,A.N.Kogan,A.D.Vikrishchuk,Russian.J.ofchem:
Vol.79,No.4,pp.826-832,2009
2.S.Prasad and R.K.Agrawal,Research letters in Inorganic Chem:10,1-4,2008
3.(Miss) K.Shrivastava and J.K.Mehrota,J.Ind.chem.soc.,38(12),1015-1017,1961
4.V.P.Sing;P.Gupta;N.Lal.Russian .J.Co-ordination chem.;34(4),270-277,2008
5.(Miss) Hil.RAY;B.S.Garg;R.P.Singh J.IND chem..soc.LVI,975-976,1979
6.S.Shit,J.Chakaraborty;B.SAMANATA;A.M.Z.Salwin,V.Gramlich;S.Mitra,Struct Chem;20,
633-642
7.S.M.Sondhi,M.Dinodia and A.kumar,Bioorg.med.chem.,14;4657,2006
8..K.Dey,and k.chakraborty,Indian.J.Chem.,39A;1140
9.A.G. Starikor, V.A. Kogan, V.V. Lukov, V.J. Minkin and R.M. Minyaev, Russian J. Co-
ordination Chem., 35(8), 616-620, 2009
10.Kedar, Ph.D. Thesis S.G.A.V. Amravati.
11.D.Pavia,G. Lampman and G.Kriz,Introduction to Spectroscopy,Thomson Asia Pvt. Lit.
Singapore,3rd Ed.,2004.
12.J.C.Hegde, N.S.Raj & Balkrishna, J.Chem.Sci. III,9(4),2007.
13.A.Hassan, A.Fetout,M.Kamal,H.A.Shraf, Molecules,10,2005.
14.R.M.Silverstein,G.C.Bassler,T.C.Morril,Spectoscopic Identification of Organic Compounds,
4th Ed.,John Wiley and Sons,INC,New York.
15.S.Chandra,and Anil Kumar,J.Saudi Chem.Soc;Vol.No.2;pp299-309,2007

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38
Study of Extraction and Chemical Screening of T. cordifolia (Guduchi)
Stem & Leaves
Ramesh Tukaram Parihar1, S A Quazi2, Shaikh Farah T3
1
Department of Chemistry,Vidnyan Mahavidyalaya Malkapur Dist.Buldhana Maharashtra-44310
2
Department of Chemistry,Bapumiya Sirajoddin Patel Arts, Commerce and Science College, Pimpalgaon kale,
Tq- Jalgaon Jamod, Dist-Buldhana
3
Department of Botany, Bapumiya Sirajoddin Patel Arts, Commerce and Science College, Pimpalgaon kale, Tq-
Jalgaon Jamod, Dist-Buldhana
Abstract:
The water & ether extracts of T.codifolia Stem & leaves were prepared- the total ash found to
be 6.861. The leaves show dark yellow fluorescent colour with picric acid the extracts were
tested for some phytochemicals. T.cordifolia commonly named as “Guduchi” is known for its
immense application in the treatment of various diseases in the traditional ayurvedic literature.
Recently the discovery of active components from the plant and their biological function in
disease control has led to active interest in the plant across the globe. Encompasses (i) the
genetic diversity of the plant and (ii) active components isolated from the plant and their
biological role in disease targeting. The future scope of the review remains in exploiting the
biochemical and signaling pathways affected by the compounds isolated from Tinospora so as
to enable new and effective formulation in disease eradication.
Keyword: T.Cordifolia, Flourescent Test, Ether Extraction, Water Extraction, Antimicrobial
activity.
INTRODUCTION:
The possibility of outbreak of severs acute respiratory syndrome (SARS) and Bird Flu Virus
continuing spread of HIV/AIDS and emergence of resistant pathogenic strains against current
medication compel investigators to look for new protective measure against these threats.
Immune activation is an effective as well as protective approach against emerging infection
disease (1). T.ordifolia commonly known as guduchi, heart-leaved moonseed and giloya is a
herbaceous vine of the family Menispermaceae, is a glabrous climbing succulent shrub. It is
native to India, easily in the tropical region. It is widely used in Ayurvedic medicine in India
as well as tonicuitalizer and as a remedy for diabetes mellitus and metabolic disorders, (2) the
fundamental role of innate immunity in host defense is becoming clearer as analysis of human
genome continues to identify new genes serving innate immune function. Innate immune
activations immune response antigen- specific T & B lympocytes (3, 4) Cytokines play crucial
roles in requlating various aspect of immune response. Among cytokines interleakin IL-12
plays a central role in co-ordinating innate and cell mediated adaptive immunity ,(5) Immune
stimulation can provide both prophylalactic as well as postexposure protection (6). Dementia
is a syndrome of failling memory and other intellectual functions with little or no disturbance
in consciousness. Degeneration of the cerbral neurons is one of the commonest and vital causes
for dementia with increasing age, there by leading to deterioration in quality of life in elderly.
Hence a greater research is required in early diagnosis of the condition and development of
newer effective drugs to prevent or halt the progression of the disease. This is possible by basic
understanding of learning and memory process (7). Medicines derived from plants have played
a pivotal role in health care of ancient and modern cultures. Ayurveda, the Indian system of
medicine mainly uses plant based drugs or formulations to treat various ailments including
cancer. Recent surveys suggest that one in three Americans uses dietary supplements daily and
the rate of usage is much higher in cancer patients, which may be up to 50% of patients treated

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in cancer centers (8). Many synthetic or natural agents have been investigated in the recent past
for their efficacy to protect against radiation damage.1 Among the natural radioprotective
agents compounds, cystine, cysteamine, 5-hydroxytryptophan, 5-hydrox- ytryptamine,
glutathione, and vitamins like A, C, and E2 have been extensively studied. Important synthetic
molecules include amino-ethyl-isothiouronium bromide hydrobromide (AET), WR-272 1.
However, the inherent toxicity of these agents at the radioprotective concentration warranted
further search of a safer and effective radioprotector. To reduce toxicily, a strategy of
combining radioprotective molecules working through different modes of action has also been
attempted.Three in fact, no radioprotective agent now available, either singularly or in
combination, meets all the requisites of an ideal radioprotector. Four recently several isolated
plant products and crude extracts that may have a natural combination of several bio-active
molecules capable of giving radioprotection through different mechanisms, have been
investigated (9).
EXPERIMENTAL: - Fresh healthy leaves and stem of T.cordifolia were collected .They were
washed thoroughly with distilled water and dried in shade for seven day followed by grinding
to make powder of the same size and stored in air tight bottles.
Total Ash: About 10 g of powdered leaves & stem was accurately weighed and taken in a silica
crucible, which was previously ignited and weighed. The powder was spread as a fine, even
layer on the bottom of the crucible. The crucible was incinerated gradually by increasing
temperature to make it dull red hot until free from carbon. The crucible was cooled and
weighed. The procedure was repeated to get constant weight.
Acid Insoluble Ash: The ash obtained as described above was boiled With 25 ml of 2N HCI
for five minutes. The insoluble ash was collected on an ash less filter paper m and washed with
hot water. The insoluble ash was transferred into a silica crucible, ignited and weighed. The
procedure was repeated to get a constant weight
Water Soluble Ash: The ash obtained as described in e determination of total ash was boiled
for 5 minutes with 25 ml of water. The insoluble matter was collected on ash less filter paper
and washed with hot water. The insoluble ash was transferred into silica crucible, ignited for
15 minutes, and weighed. The procedure was repeated to get a constant weight. The weight of
insoluble matter was subtracted from the weight of the total ash. The difference of weight was
considered as water-soluble ash the result of total ash, acid insoluble ash, water soluble ash and
other physical Para meters of T.cordifolia leaves are summarized in Percentage of ash and
Chemical Screening.
Antimicrobial Activity: Antimicrobial activity of T.cordifolia plant was determined against
bacterial strain, salmonella typnis Escherichia Coil, Staphy Lococcus acereus Bacillus subtilis
by well diffusion assay on agarr plate.The bacterial culiure were grown on nutrient broth for
24 hrs. The activity grown cullirs were spread on nutrient agar plates by spread plate method
well were prepared by brose sample was poured in the well. Streptomycin antibiotic is used as
standard 100 mg 1 ml concentration.
WATER EXTRACTION: 10 gm of sample (Tinospora plant) was taken in round bottom
flask. 35ml of distilled water was added. Water condenser was arranged. Refluxed for 3 hrs
after complete heating the water extract was cooled. Solution was filtrated throught whatman
paper No. 42 Residue was dried and weight was took and water soluble compound was
calculated.
ETHER EXTRACTION: 10 gm of leaves & stem sample was taken Tinospora plant (10 gm
each) was extracted using Round bottom flask, soxhlet apparatus was arranged. The extracts

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were too dried to yield crude residue. The extracts were auto-calved and stored at 50c until
further use. Ether soluble compound was calculated.
RESULT & DISCUSSION :In study of three crude extracts of leaves and stem of
T.Condifolia have been investigatged, aqueous either and water extraction were tested for their
total Fluonaid contents microbial activity performed by using assay of reducing powder of
Tinospora plant.
Percentage of ash
Sr. No. Sample (ash) Percentage
1. Total Ash 5.78%
2. Water Soluble 28.03%
3. Acid Insoluble 59.00%

Chemical Screening
Sr. No. Chemical Screening Water Ether
Extraction Extraction
1. Tannins +ve -ve
2. Saponins -ve +ve
3. Phenols +ve +ve
4. Flavonoids -ve -ve
5. Alkaloids -ve +ve

REFERENCE:
1. P.K. Raveendra Nair, Sonia Rodiguez, Reshma Ramachandran, Arturo Alamo, Steven J. Melnick,
Enrique Escalon & Pedro I, Immune stimulating properties of a novel polysaccharide from the
medicinal plant Tinospora Cordifolia. International Immunopharmacology, 4,(2004),1645-1659.
2. P. Stanely, Mainzen Prince & Venugopal P. Menon,Hypoglycaemic and other related actions of
Tinospora conrdifolia roots in alloxan-induced diabetic rats. Jounral of Ethnopharmacology,
70,(2000) ,9-15.
3. Salah. M. EL Naggar, Botany Dept. Pollen Morphology of Egyptian Malvaceae: Abutlion
pannosum .28,(2004),227-240.
4. Anup A. Arbat, Pharmacognostic Studies Stem of Tinospora Plant, BioscienceDiscovery,3 (3),(
2012),317-320.
5. Survase S.A., B.P. Sarwade and D.P. Chavan Antimicrobail Activity of various extracts of
Tinospora Plant, African Journal of Plant Science. 7 (4),(2013),128-130.
6. Walker P.S., Scharton-Kersten T., Krig A.M., Homant, Rowton E.D. & Udey M.C. at
Immunostimunity oligodeox-yucdeotide s. protective immunity and provide systemic terepy for
leishmaniasi, IL-12 and IF N - gamma depend + mechanism .96,(1999),6970-6957.
7. Ashutosh Agarwal, S. Malini, K.L. Bairy, Muddanna S.Rao. Effect of Tinospora Cordifolia on
Learning and Memory in Normal and Memory Deficit Rats. Indian Journal of Pharmacology .
34,(2002),339-349.
8. Ganesh Chandra Jagetia and Shaival Kamalaksha Rao, Evaluation of the Antineoplastic Activity of
Guduchi (Tinospora Cordifolia) in Ehrlich Ascites Carcionma Bearing Mice. Biol. Pharm, Bull
29,(3),(2006) ,460-166 .
9. Harish Chandra Goel, Jagadish Prasad, Surinder Singh, Ravinder Kumar Sagar, Paban Kumar
Agrawala, Madhu Bala, Auran Kumar and Ruchi Dogra, Radioprotective Potendial of an Herbal
Extract of Tinospora Cordifolia, 45, (2004), 61-68.

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39

In-silico QSAR-based virtual screening recognition of novel anaphylactic

lymphoma kinase inhibitor

Rahul D. Jawarkar 1*, Praveen Sharma1, Sachin Kumar Jain1

1
Faculty of Pharmacy, Oriental University, Indore 453555, Madhya Pradesh, India;
*Contact: [email protected]; Mobile: +91 7385178762.

Abstract: The goal of this work is to find a new treatment option that blocks ALK tyrosine
kinase. We used drug repositioning and a QSAR-based virtual screening method together in
this work to find a new lead drug candidate that might be able to stop ALK tyrosine kinase
receptors. Molecular docking, MD modelling, and MMGBSA studies were used to look into
how the new FDA-approved drugs interact with each other. To make sure the results we got
online were correct, we used the A549 lung cancer cell line to do an in vitro MTT test. The
usual method suggested by the OECD is used in this study. It includes QSARINS GA-MLR
models, QSAR-based virtual screening of about 1652 FDA chemicals, MD simulation, and
MMGBSA analysis using Desmond software. The MTT test confirmed what was found in the
computer model. The QSAR model that was made meets a number of validation standards,
such as R2 = 0.79, Q2LOO = 0.78, Q2LMO = 0.78, R2ex = 0.77, and CCCex = 0.87. Using QSAR-
based virtual screening, we also found 12 FDA drugs that were hits in the computer. Some of
these substances could be used as ALK-TK inhibitors in clinical settings because their docking
values ranged from -7.10 to -10.57 kcal/mol. The A549 cancer cell line was used in the MTT
test to support the results that were found in a computer programme. The QSAR-based virtual
screening indicated that the new molecule ZINC000150338819 would have a PIC50 of 9.18 M.
This drug has both wild-type and mutant ALK-TK. The molecule got a docking score of -10.57
kcal/mol and an RMSD of 1.54Å as well. The ZINC000150338819-ALK TK complex is stable,
and it includes both wild-type and mutant ALK TK. This was proven by MD modelling and
MMGBSA tests that lasted 200 ns. To confirm the results from the computer simulation, the
MTT test shows that Ledipasvir had more inhibition than ceritinib. It is suggested in this study
that the chemical ZINC000150338819 could be used as an ALK TK inhibitor in the drug
development field.

Keywords: OECD; GA-MLR; MD simulation; MMGBSA; X-Ray.

1 Introduction
In 1991, researchers discovered a membrane-bound tyrosine kinase receptor called anaplastic
lymphoma kinase (ALK). Abnormal forms of ALK, such as fusion proteins, point mutations
that activate ALK, and gene amplification, are found in cancer. Abnormal ALK expression is
linked to the development of many types of cancer [1]. The human ALK gene is located in the
chromosomal region 2p23.2p23.1. This 26-exon gene encodes the ALK protein, which is 1620
amino acids long. The full-length ALK protein has a transmembrane region, a ligand-binding
region, and an intracellular tyrosine kinase region consisting of 561 amino acids[2,3]. The 3-
tyrosine motif (Tyr1278, Tyr1282, and Tyr1283), where autophosphorylation for kinase
activity occurs, is also found in other members of the same family of kinases (See fig.1).

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.
Figure 1 Structure of ALK, showing a polypeptide of 1620 amino acids. The full-length form
of ALK resembles other receptor tyrosine kinases. Kinase activity is regulated by a 3-tyrosine
motif (Tyr1278, Tyr1282, and Tyr1283), which is located in an intracellular tyrosine kinase
domain.

Figure 2 Some of the ALK tyrosine kinase discovered till date and available in clinical practice.

Several research studies have revealed that TKIs' ability to inhibit ALK kinase activity has a
substantial anticancer effect [4-11]. Moreover, numerous potent and selective ALK-TKIs have
been designed to inhibit fusion proteins and activate ALK variants[12-22]. Targeted therapy
using the epidermal growth factor receptor (EGFR) has been discovered as a treatment for
NSCLC patients with ALK activation mutations. However, most patients who receive targeted
treatment relapse due to genetic changes that confer resistance[12,23]. Many ALK TKIs are
available for cancer therapy in different regions. The FDA has approved several ALK inhibitors
for cancer therapy, including ceritinib, brigatinib, crizotinib, alectinib, TPX-0131,
GSK1838705, CEP28122, AP26113, and X-396 (See fig. 2).
In the present work, we used a QSAR-based virtual screening approach combined with drug
repositioning to find a unique lead drug candidate that might effectively inhibit ALK tyrosine
kinase receptors. The interactions between the newly discovered FDA drugs were investigated
using molecular docking, MD modelling, and MMGBSA studies. To corroborate the results
obtained virtually, we conducted an in vitro MTT experiment using the A549 lung cancer cell
line.

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2 Material and Method

2.1 QSAR Methodology

The current study adheres to the conventional technique advised by the OECD and other
scholars for doing QSAR analysis[24-26]. All tools were used with their default values;
however, certain settings were altered, and these are detailed in the documentation, in order to
generate a robust QSAR model with an equilibrium of predictive capacity and mechanistic
understanding.

Step 1: Data collection and curation:

To start, the Binding DB (https://www.bindingdb.org/bind/index.jsp; last accessed:
12/24/2021) was used to get a large dataset with 1806 IC50 values for experimentally proven
ALK-TK inhibitors. Data quality, and suitable curation before further processing have a
significant impact on QSAR analysis[25,27-29]. After that, we filtered the data[30], which
included eliminating duplicates, organometallic compounds, salts, molecules with ambiguous
IC50 values, and so on. This resulted in a reduction of the dataset's molecules from 1807 to
1328. In spite of the reduced size of the dataset, it still included molecules with experimental
IC50 (nM) values between 0.3 and 83,000 nM and the occurrence of different scaffolds such as
heterocyclic rings, positional isomers, stereoisomers, etc., all of which widened the chemical
space and increased the model's applicability. Table 1 (See Table 1 in the Supplementary
Materials) contains the SMILES (Simplified Molecular Input Line Entry System)
nomenclature for all of the compounds used in this investigation, together with their
experimental IC50 and pIC50 (=log10IC50).
Table 1 shows a few representative values for IC50 (nM) and pIC50 (M) in the SMILES format,
along with some examples of the most and least active compounds.

Table 1. SMILES notation, IC50 (nM), and pIC50 (M) values for the five most and least active
compounds in the selected data set.
IC50
Id SMILES in nm PIC50

25 CCc1cc2C(=O)c3c([nH]c4cc(ccc34)C#N)C(C)(C)c2cc1N1CCC(CC1)N1CCC(O)CC1 0.3 9.523

33 C[C@H]1Oc2nc(cnc2N)-c2cc(ccc2CN(C)C(=O)c2ccc(F)cc12)S(C)(=O)=O 0.34 9.469
35 COc1cc(ccc1Nc1ncc(Cl)c(Nc2ccccc2P(C)(C)=O)n1)N1CCC(O)CC1 0.36 9.444
36 COc1cc(ccc1Nc1ncc(Cl)c(Nc2ccccc2P(C)(C)=O)n1)N1CCC(CC1)N1CCN(C)CC1 0.37 9.432
37 CCN1CCCc2cc(Nc3ncc(Cl)c(Nc4ccccc4S(=O)(=O)C(C)C)n3)c(OC)cc2C1 0.38 9.42
1791 CN1CCN(CC1)c1ccc(Nc2ncc3ccc(-c4ccccc4C(N)=O)n3n2)cc1 726 6.139
1793 COc1ccccc1C#Cc1ccnc2[nH]c3ccc(cc3c12)-c1ccc(cc1)N1CCN(C)CC1 750 6.125
1800 CN1CCN(Cc2ccc(cc2)C(=O)Nc2nc(cs2)-c2ccc(C)c(c2)C#Cc2ccc(C)cc2)CC1 83000 4.081
1803 CC1(C)c2[nH]c3cc(ccc3c2C(=O)c2ccc(OC[C@H](O)CO)cc12)-c1ccn[nH]1 770 6.114
1805 CC(C)S(=O)(=O)c1ccccc1Nc1nc(Nc2nc3CCN(C)CCc3s2)ncc1Cl 773 6.112

Step-2 Second, we utilised the default settings for OpenBabel 2.4[31] and MOPAC 2012
(openmopac.net, obtained on March 5, 2022) to generate SMILES notations for the optimum
3D structures of the compounds (semi-empirical PM3 technique).
Step 3: If enough molecular descriptors are generated and subsequently pruned to limit the
chance of overfitting from redundant noisy descriptors, then a QSAR model may strike a
satisfactory balance between mechanistic interpretation and predictive ability[32]. Then,
molecular descriptors were generated from 1D to 3D for every molecule. For this objective, we
used PyDescriptor [33], which can calculate over 40,000 molecular descriptors for a given

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molecule. To reduce the number of molecular descriptors in the descriptor pool, we used
QSARINS 2.2.4 to eliminate duplicates and variables with strong correlation (|R| > 0.95 or >
98%) [34]. This resulted in a reduction from 40,000 to 2,376, but still covered a broad spectrum
of molecular descriptors.

2.2 Subjective Feature Selection (SFS) involves separating the dataset into a training set
and an external set
Splitting the dataset into a training set and an external set (also known as a prediction set or
test set) is essential for developing and validating a reliable QSAR model [25,27-29]. In order
to exclude any possible bias, we divided the dataset into a training set of 1062 molecules (80%)
and an external collection of 266 molecules (20%) for this study. The only purpose served by
the external set was model validation (predictive QSAR), whereas the molecular descriptors
were selected from the training set to determine how many parameters were desired. QSARINS
2.2.4 was used to develop the model through multi-linear regression (MLR) and the Genetic
Algorithm (GA). With a fitness function of Q2LOO and 10,000 iterations. Determining the right
number of molecular descriptors to use in a model's creation is an important step in quantitative
structure-activity relationship (QSAR) modelling. Until the value of Q2LOO rose beyond a
certain threshold, the heuristic search had to construct a large number of models, beginning
with a univariate model and progressing to a multivariate model as additional molecular
descriptors were included.

Figure 3 Graph for the number of descriptors vs. leave-one-out Coefficient. Q2LOO for optimal
descriptor count.

Figure 27 is a two-dimensional graph showing the correlation between the Q2LOO values and
the number of chemical descriptors employed by the models. Adding more molecular
descriptors did not increase the model's statistical performance; hence, this threshold was used
to determine the ideal number of variables to include in the model. The investigation
established critical values for six independent variables[35,36] (See Fig. 3). This meant that
the QSAR models with more than six descriptors had to be rejected.

2.3 Constructing a Valid Regression Model and its validation

Several validation methods, such as cross-inter validation, external validation, Y-
randomization analysis, and the applicability domain (William's plot), can be used to test how

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accurate and reliable a QSAR model is. A well-validated QSAR model is very helpful for
virtual screening, lead/hit optimisation, decision-making, and other activities. Several types of
validation—cross-inter, external, Y-randomization, and applicability domain (William's
plot)—can be used to approximate the robustness and wide application of a QSAR
model[25,37-40].
The following are the typical criteria for assessing QSAR models, together with their
corresponding threshold values for validation parameters[35,36].There have been several
successful model generation efforts using GA-MLR. The best possible model was selected
using the following stringent parameters and criteria: R2tr ≥ 0.6, Q2loo ≥ 0.5, Q2LMO ≥ 0.6, R2
> Q2, R2ex ≥ 0.6, RMSEtr < RMSEcv, K ≥ 0.05, CCC ≥ 0.80, Q2-Fn ≥ 0.60, r2m ≥ 0.5, (1-
r2/ro2) < 0.1, 0.9 ≤ k ≤ 1.1, or (1-r2/r’o2) < 0.1, 0.9 ≤ k’ ≤ 1.1,| ro2-r’o2|< 0.3, RMSEex,
MAEex, R2ex, Q2F1, Q2F2, Q2F3, and low R2Yscr, RMSE and MAE.
Validation of quantitative structure-activity relationship (QSAR) models entails establishing
the model's applicability. We used a Williams plot to determine the extent to which the QSAR
model was reliable. (Table 2 in the supplemental materials lists the computed descriptors used
to create the QSAR model.) Table 3 of the supplementary materials provides the formulas for
determining these statistical features.

2.4 QSAR-Based Virtual Screening for Drug Repositioning (Repurposing)

Due to the rising need for novel anticancer treatments, drug repurposing has fascinated the
cancer research community. Even though there are many ways to treat cancer, such as
chemotherapy and targeted therapies, cancer is defined by its inability to respond to current
medicines and drugs. Because of this, studying potential novel cancer treatments is a thriving
field of study[41-43]. Nowadays, drug repositioning is an emerging area of study; therefore,
we utilised this information to conduct a QSAR-based virtual screening using the zinc database
and its 1615 FDA compounds. Since then, QSAR-based VS has used 1615 FDA substances.
Before doing molecular descriptor calculations, the 3D structures of molecules were
constructed in the same way as a modelling set. The ALK-TK inhibitory action of 1615 FDA-
approved drugs was predicted using a completely validated six-parameter quantitative
structure-activity relationship (QSAR) model, which was derived using estimated chemical
descriptors. The chemical details and predicted IC50 values for the zinc FDA 1615 compounds
are included in Table 4 of the supplemental materials.

2.5 Molecular Docking Analysis

The ALK TK wild type (pdb-4cmu) and mutant (pdb-4clj) Protein Data Bank (pdb) data were
obtained from the protein data bank[44]. The pdb:4cmu and 4clj were chosen on the basis of
X-ray resolution and completion of amino acid sequences. Ramachandran's plot was used to
determine the protein's health before running docking simulations. The protein, after
optimisation, passes muster for docking studies. Both PDB files had their native ligands
removed before docking analysis could begin. To facilitate comparison between the wild-type
(4cmj) and mutant (4clj) strains, all 12 hit molecules from the QSAR-based virtual screening
were docked into the active sites of both. For convenience, we've included the docking position
of Ledipasvir, the most active molecule.
Molecular docking analysis was performed in NRGSuite software package [45]. This is an
open-access tool that is available at no cost as a plugin for the PyMOL software
(www.pymol.org as of March 9, 2022). FlexAID can help you find cavities on protein surfaces
to ensure that it can be utilised in docking simulation targets[46]. Covalent docking,
conformational search using a genetic algorithm, and the mobility of ligands and side chains
are all modelled. For optimal performance in this study, NRGsuite was run with the following
flexible-rigid docking with default parameters: Input Method for Boundary Sites: HET groups

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contain water molecules; have a van der Waals permeability of 0.1; have gone through 1000
generations; use the share fitness metric; reproduce using the population explosion model; have
five TOP complexes; have a cylindrical form (diameter: 19); have a three-dimensional grid
spacing of 0.367; have no side-chain mobility; be ligand-adaptable; not have a ligand posture
for comparison; have no constraints; and so on. For both the wild-type and mutant strains, the
accuracy of molecular docking was tested using two molecules: PF-06463922 and (10R) -7-
amino-12-fluoro-1, 3, 10, 16-tetramethyl-16, 17-dihydro-1H-8, 4-(metheno) pyrazole (4, 3-)
(2, 5, 11). ((10R) -7-amino-12-fluoro-2,10,16-trimethyl-15-oxo-10,15,16-tetrahydro-2H-8) 4-
(metheno) pyrazole (4, 3-h) (2,5,11) is a well-characterised inhibitor of ALK TK that was used
to verify the docking method.
2.6 Molecular dynamics (MD) simulations
Molecular dynamics and simulation (MDS) methods were used to look into how stable and
convergent the interaction between ledipasvir and ALK TK was. This study examined both
wild (pdb-4cmu) and mutant (pdb-4clj) strains for their stability. The system builder was used
to construct intricate systems for the strains of Ledipasvir-wild, Ledipasvir-mutant, Ceritinib-
wild, and Ceritinib-mutant. This action was undertaken in order to facilitate the execution of
simulations. The system used the OPLS-2005 force field and included an explicit solvent model
using SPC water molecules[47,48]. The baseline parameters for the explicit SPC water model's
orthorhombic box measuring 7.0 x 7.0 x 7.0 metres were established using Desmond 2018-
4[49]. The neutralisation of both wild-type and mutant ALK-TK complexes was achieved by
introducing NaCl salt at a concentration of 0.15 M Na+ ions. In the Desmond system builder
panel, the neutralise option has been selected to introduce a predetermined quantity of
counterions. In the molecular dynamics simulation approach, 18 sodium (NA) ions and 15
chloride (Cl) ions were included. After using the ASL module to choose certain residues of the
ligand and protein molecules, the Desmond default relaxation technique was employed to
improve the performance of the resulting systems. To learn more about each complex, we
conducted molecular dynamics (MD) simulations. In the previous production run, we kept the
temperature and pressure (NPT) constant and ran a molecular dynamics simulation (MDS) for
200 ns. The Nosé-Hoover chain coupling method was used to create the NPT ensemble, and
the final simulation was run at 300 K with a relaxation period of 1 ps throughout the whole
dynamics [49,50]. With a relaxation time of just 2 picoseconds[51], pressure was controlled
using a barostat based on the Martyna Tuckerman-Klein chain coupling system. The Desmond
simulation used the isotropic Martyna-Tobias-Klein barostat and the Nose-Hoover thermostat
to regulate the pressure at 1 atmosphere and the temperature at 300 Kelvin. The NPT ensemble
was used in all runs, with a temperature of 300 K and a pressure of 1 bar. We successfully
estimated the bonding interactions by utilising a time step of 2 femtoseconds and the RESPA
integrator. Using the particle mesh Ewald method, and keeping the radius for Coulomb
interactions at 9 [52], we were able to calculate the long-range electrostatic interactions
between the particles. This investigation describes the remaining possible setups. After
finishing the last simulation run, the simulated trajectories of the wild-type and mutant
Ledipasvir strains were analysed. Root-mean-square deviation (RMSD), root-mean-square
fluctuation (RMSF), and hydrogen-bond formation were the primary areas of study in this
examination. Binding energies for the complexes were estimated using the MM-GBSA
technique, which was applied to 200 individual 1 ns trajectories. Standard deviations and mean
binding energies were calculated from the data so obtained.

2.7 Molecular Mechanics: Generalised Borne Surface Area

Docked complexes of ledipasvir and ceritinib were analysed to determine their binding free
energy (Gbind) with the help of the MM-GBSA module. During molecular dynamics (MD)
simulations, the ALK complex was attached to both natural (4cmu) and mutant (4clj) strains,

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allowing for this estimate to be made. The New York-based Schrodinger Suite, LLC, version
2023–24, was used to run the simulations. Binding free energy was determined using a rotamer
search and calculated with the OPLS 2005 force field and the VSGB solvent model [53]. After
an MD experiment was completed, a time window of 10 ns was used to choose the frames of
the trajectories. By using Equation 1, the comprehensive free energy of binding was
successfully determined.

ΔGbind = Gcomplex – (Gprotein + Gligand) (1)

Where,
ΔGbind = binding free energy,
Gcomplex = free energy of the complex,
Gprotein = free energy of the target protein,
and Gligand = free energy of the ligand.
The trajectories of the MMGBSA results were analysed to learn more about the structural
changes that occurred after the dynamics were applied.

2.8 In-Vitro Evaluation of Anticancer Activity by MTT Assay

The A549 lung cancer cell line, at passage 68, was purchased from NCCS, Pune, India. F-12K
medium, antibiotic-antimycotic solution, HEPES solution, and 10% foetal bovine serum were
used to cultivate the cells after they were frozen. The experiment used the 3(4,5-dimethyl-
thiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay to assess mitochondrial function.
This assay relies on the reducing properties of MTT, which lead to the formation of insoluble
formazan crystals specifically inside viable mitochondria. In summary, 1x104 A549 cells were
seeded into each well of a 96-well plate, and the plate was then incubated for 24 hours at 37°C
in 5% CO2. Following removal of the medium, the cells were treated with Ceritinib (5 M),
Posaconazole (10 M), Ledipasvir (20 M), and Ledipasvir (40 M) for 24 hours in triplicate. The
chemical ceritinib was used as a standard. Each well was given a media volume of 300 l. After
24 hours of treatment, 25 L of MTT solution (5 mg/mL) was added to each well, and the cells
were incubated for 4 hours at 37°C in a 5% CO2 atmosphere. Following the dissolution of the
formazan crystals in a volume of 100 litres of dimethyl sulfoxide (DMSO), the absorbance was
then determined at a wavelength of 570 nanometers using an Epoch Microplate
Spectrophotometer manufactured by Biotek Instrument. The IC50 values were calculated using
GraphPad Prism (version 7) software. Examining a nonlinear plot of the percentage of cell
inhibition against the logarithm of concentration enabled this. The calculation of cell growth
inhibition percentage was performed using the below formula:

% Cell Viability = (AT / AU) X 100

Where; AT = Absorbance of Treated Cells (Drug)

AU = Absorbance of Untreated Cells, % Cell Inhibition = 100 - % Cell Viability

3 Results and Discussion

This work used molecular docking and quantitative structure-activity relationship analysis to
identify ALK-TK inhibitory structural components. A QSAR paradigm combines structural
characteristics to basic chemical descriptors. The six-parametric GA-MLR model's structural
interpretation and clear molecular descriptors make it a good external predictor. IC50 values of
molecules in the dataset can explain the effect of a specific descriptor, but the combined or

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inverse effect of unknown factors or other molecular descriptors could have a significant
impact on a molecule's IC50 value (See figure 4).
QSAR Model
PIC50= 5.577 (± 0.087) + 1.21 (± 0.102) * aroC_sumpc + 0.086 (± 0.006) * ringC_plaN_6B
+ 0.221 (± 0.029) * fnotringNsp3C4B + -0.269 (± 0.027) * faroNC8B + 1.059 (± 0.103) *
fdonnotringN5B + -0.595 (± 0.061) * fnotringNringN4B+

R2:0.7909, R2adj:0.7897, R2-R2adj:0.0012, LOF:0.2544, Kxx: 0.3058, Delta K:0.0661,

RMSEtr: 0.4987, MAEtr: 0.4215, RSStr: 264.3315, CCCtr: 0.8832, s: 0.5003, F: 665.6663,
Q2loo: 0.7882, R2-Q2loo: 0.0027, RMSEcv: 0.5019, MAEcv: 0.4242, PRESScv: 267.7734, CCCcv:
0.8817, Q2LMO: 0.7885 ,R2Yscr:0.0056, Q2Yscr:-0.0076, RMSEAVYscr:1.0874, RMSEext:0.5285,
MAEext:0.4460, PRESSext:74.0260, R2ext:0.7710, Q2-F1:0.7708, Q2-F2:0.7690, Q2-F3:0.7651,
CCCext:0.8739, r2m aver.:0.6744, r2m delta:0.1472, R2: 0.7882, R'2o: 0.7334, k': 0.9956, Clos':
0.0695, r'2m: 0.6038. Pred(x) vs. Exp(y): R2: 0.7882, R2o: 0.7882, k: 1.0000, Clos: 0.0000, r2m:
0.7869, Exp(x) vs. Pred(y): R2: 0.7710, R'2o: 0.7223, k': 0.9998, Clos': 0.0632, r'2m: 0.6008,
R2: 0.7710, R2o: 0.7701, k: 0.9951, Clos: 0.0012, r2m: 0.7480.

Figure 4 QSAR model development and accompanying graphs (a) A scatter plot contrasting
the expected and experimental PIC50 values (b) a portrayal of a Williams plot to assess the
model's applicability domain; and (c) a presentation of an Insubria plot.

3.1 QSAR Mechanistic Interpretation

The significance of the sum of partly charged carbon atoms in aromatic fragments is shown by
aroC_sumpc. It is one of the variables that exhibit a positive correlation in the constructed
QSAR model. These descriptions highlight the charges, both complete and partial, that are
linked to carbon atoms with aromatic and fragrant properties. The presence of a positive
coefficient for this descriptor suggests that there is a direct relationship between a greater
activity profile and a higher aroC_sumpc score. The evaluation involves analysing the
following pair of molecules: 502 (502, IC50=4.67 nM, aroC_sumpc=0.65,
ringCplus_sumpc=0.97) and 1030 (1030, IC50=19.05 nM, aroC_sumpc=0.49,
ringCplus_sumpc=0.85)( see fig 5).

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Figure 5 The molecular descriptor aroC_sumpc, shown exclusively for molecules 502 and
1030.

Ring carbon and planar nitrogen were found to be important for TK ALK to work as an
inhibitor (ringC_plaN_6B). It shows that there are ring carbons in the six bonds of the flat
nitrogen atoms. It's much better for TK-ALK to stop a reaction when the positive factor number
for this trait in the QSAR model goes up. One way to show this is to compare molecule 1065
(1065, IC50 = 21.38 nM, ringC_plaN_6B = 20, C_plaN_5B = 26, aroC_plaN_6B = 16) to
molecule 1778 (1778, IC50 = 69183 nM, ringC_plaN_6B = 16, C_plaN_5B = 16,
aroC_plaN_6B = 12) (See Fig. 6)

Figure 6. For molecules 1065 and 1778, the molecular descriptor ringC_plaN_6B is shown.

fnotringNsp3C4B: The abbreviation fnotringNsp3C4B describes how often Sp3-hybridized

carbon atoms are found within 4 bonds of acyclic or non-ring nitrogen atoms. Since it has a
positive coefficient, the value of the molecular marker fnotringNsp3C4B goes up as the activity
profile does. The fnotringNsp3C4B is not worked out if the same sp3 hybrid carbon atom is
three or five bonds away from the carbon atom. This is clear when you look at the molecule
135 (IC50 = 1 nM, fnotringNsp3C4B = 2, fnotringNnotringC4B = 2, fplaNsp3C4B = 2) next to
the molecule 1670 (IC50 = 371.5 nM, fnotringNsp3C4B = 1, fnotringNnotringC4B = 1,
fplaNsp3C4B = 1) (See fig. 7).

Figure 7: For molecules 135 and 1670, the fnotringNsp3C4B molecular descriptor is shown.
fdonnotringN5B (The chance of a nitrogen atom that is not in a ring is exactly 5 bonds away
from the source atoms.) This trait has a positive coefficient in the standard QSAR model. This
means that as its value goes up, ALK-TK suppression goes up as well. It can be seen that
between molecules 245 (IC50 = 2.13 nM, fdonnotringN5B = 1) and 1449 (IC50 = 812.8 nM,
fdonnotringN5B = 0). This might explain the differences in inhibitory activity of ALK TKs
(See Figure 8).

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Figure 8 For molecules 245 and 1449, the molecular descriptor fdonnotringN5B is shown.
The abbreviation faroNC8B shows how often aromatic nitrogen atoms with carbon atoms
connected in exactly 8 bonds are found. The suggested QSAR model shows that this trait has
a negative coefficient. Increasing its value would make ALK TK even less effective at blocking
it. To make ALK TK as good at stopping cell growth as possible, more changes should be made
to the molecule that lower the value of faroNC8B. This remark can be explained by comparing
molecule 1168 (IC50 = 107.1 nM, faroNC8B = 2) to molecule 1118 (IC50 = 81.8 nM, faroNC8B
=1) (see fig. 9).

Figure 9 Illustration of the molecular descriptor faroNC8B for the molecules 1168 and 1118.

fnotringNringN4B (frequency of occurrence of a ring nitrogen atom exactly at 4 bonds from

a non-ring nitrogen atom) As the value of the descriptor goes up in the QSAR model that was
made, negative coefficients of the descriptor make the inhibitor less active. This can be
demonstrated by comparing molecule 192 (PIC50 = 8.77, fnotringNringN4B = 0) with molecule
1343 (PIC50 = 6.43, fnotringNringN4B = 1) (see fig. 10).

Figure 10 The fnotringNringN4B molecular descriptor is presented only for molecules 192
and 1343.

3.2 Drug Repositioning and QSAR Based Virtual Screening

After making the QSAR model, we used it to predict the ALK-TK inhibitory activity of 1650
FDA molecules through a QSAR-based virtual screening. The 12 hit molecules were obtained
as repurposed drug candidates against the ALK-TK receptor. Based on its PIC50, the molecule
ZINC000150338819 (Ledipasvir) was chosen as a major hit among the top hit molecules.

3.4 Applicability domain study of the Identified hit molecules

To see how widely our QSAR model could be used, we used a collection of 1329 molecules
for training and a set of 12 hit molecules for forecast. Twelve molecules were found through
QSAR-based virtual screening and were then described. As you can see in Fig. 11, the hit
molecules ZINC000150338819, ZINC000150588351, ZINC000203686879, and

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ZINC000068204830 are on the edge of the application domain in the Williams plot. This means
that the leverage value is low.

Figure 11 The 12 hit molecules from the QSAR-based virtual screening are shown in a
Williams plot for their potential applicability domain (blue dots represent the hit molecules).

3.5 Molecular Docking analysis

All reported hit compounds were docked to ALK TK wild (pdb-4cum wt) and mutant strains
(pdb-4clj mutant) to analyse the binding relationships. The QSAR-based virtual screening
anticipated several FDA compounds' ALK-TK inhibitory activity. The docking scores and
predicted activity (PIC50) of the 12 hit molecules for the wild-type ALK TK and the mutant
L1196M TK are shown in Tables 1 and 2, respectively. The QSAR-based virtual screening
showed that all antiviral drugs had a greater PIC50 against both wild and mutant ALK TK strains
than the clinically proven ALK TK inhibitors (See Tables 2 and 3).

Figure 12 Depiction of 2D interaction) of ZINC000150338819 (Ledipasvir) with ALK TK

wild strain (A) and mutant strain(B).

Table 2 Depiction of the Docking Results of the 12 hits for ALK TK (pdb-4cmu, wild strain)
PIC50 by Docking Binding
RMSD
SN ZINC I.D. hit molecules QSAR Score free
Å
VS kcal/mol energy
1 ZINC000150338755 (Venetoclax) 9.36 -9.90 1.64 -38.63
2 ZINC000150338819(Ledipasvir) 9.18 -10.57 1.54 -55.19
3 ZINC000150588351(Elbasvir) 9.02 -9.93 2.37 -61.26
4 ZINC000066166864 (Alectinib) 8.67 -7.65 1.96 -34.99
5 ZINC000203686879 (Velpatasvir) 8.58 -10.08 2.10 -51.97

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6 ZINC000148723177(Brigatinib) 8.53 -8.45 1.78 -29.96

7 ZINC000096272772(Ceritinib) 8.28 -8.17 1.58 -38.64
8 ZINC000028639340 8.25 -8.94 2.65 -50.07
9 ZINC000003938482(Posaconazole) 8.25 -9.70 1.59 -49.49
10 ZINC000072316335(Ribociclib) 8.20 -7.48 1.12 -34.62
11 ZINC000068204830(Daclatasvir) 8.14 -10.10 2.15 -49.15
12 ZINC000003787097(Besifloxacin) 8.03 -7.10 0.99 -32.95

Figure 13 Depiction of 2D interaction) of ZINC000096272772 (Ceritinib) with ALK TK wild

strain (A) and mutant strain(B).

Table 3 Depiction of the docking Results of 12 hits for ALK TK (pdb-4clj, mutant strain).
PIC50 by Docking
QSAR Score Binding free
sn ZINC I.D. VS kcal/mol RMSD A0 energy
1 ZINC000150338755 (Venetoclax) 9.36 -8.87 2.28 -46.77
2 ZINC000150338819(Ledipasvir) 9.18 -8.52 2.09 -37.33
3 ZINC000150588351(Elbasvir) 9.02 -8.95 4.25 -39.68
4 ZINC000066166864 (Alectinib) 8.67 -7.70 1.47 -32.47
5 ZINC000203686879 (Velpatasvir) 8.58 -9.35 1.70 -31.60
6 ZINC000148723177(Brigatinib) 8.53 -8.33 1.70 -40.17
7 ZINC000096272772(Ceritinib) 8.28 -8.36 1.60 -34.64
8 ZINC000028639340 8.25 -7.07 2.19 -23.41
9 ZINC000003938482(Posaconazole) 8.25 -8.29 3.46 -44.94
10 ZINC000072316335(Ribociclib) 8.20 -7.93 3.70 -39.19
11 ZINC000068204830(Daclatasvir) 8.14 -8.43 4.77 -41.96
12 ZINC000003787097(Besifloxacin) 8.03 -7.27 1.89 -30.17

It has a strong preference for Ledipasvir because the ALK TK mutant strain reacts with it in a
way that is less water-friendly than the normal ALK TK. Arg1253 formed a normal hydrogen
bond with Ledipasvir, and water formed hydrogen bonds with HOH2151, HOH2155, and
HOH2151. It takes -0.33 kcal/mol of energy for ledipasvir to bind to Met1199 in the active
"DFG-in" shape of wild-type ALK TK. It needs -0.88 kcal/mol in the mutant strain. This shows
that Ledipasvir has a stronger attraction for the mutant ALK TK strain compared to the wild
strain, even though the contact distance between the two strains was only 3.88. The gap
between Gly1202 and fluorine atoms that made touch was found to be 4.14. In the wild ALK
TK, which was shaped like a boat, no similar contact was found between Gly1202 and fluorine
atoms. Also, the way the fluorine atoms are arranged in the hydrophobic pocket brings them

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closer to the Asn1254 residue of the mutant ALK-TK. This improves the interactions between
molecules and keeps its chair-shaped shape. Along the same lines, this is like how fluorine
atoms are oriented towards the Asn1254 residue of the pdb-4clj binding. The difference in
binding free energy between the two strains could be because Ledipasvir has different shapes
when it comes to normal and mutant ALK TK. Also, the wild ALK TK had a fluorine atom
arrangement with a dihedral angle of 52.2 degrees. The mutant ALK TK, on the other hand,
had an angle of 70 degrees, which means it rotated 18 degrees differently. This shows that the
mutant ALK TK goes through big changes in its shape that affect its capacity to attach to
Ledipasvir (see fig 112 and 13).

3.6 Molecular dynamics (MD) simulations

The apo-ALK tyrosine kinase (TK) was stable throughout the 200-nanosecond simulation,
except for a 60–85 nanosecond variation. The c-alpha atoms' root mean square deviation
(RMSD) varied from 2.0 to 2.2 angstroms at this time. The system converged at 2.27 angstroms
RMSD. The apo-ALK tyrosine kinase (TK) showed larger variations in the back area (RMSD
= 2.3 Å) and sidechain region compared to the C- and backbone regions (RMSD = 3.2) (see
figure 1(B)). Figure 1 indicates that the wild type A-loop RMSD converged to 1.8 after 40 ns
of simulation. In the L1196 mutant, the A-loop RMSD reached 2.4 at 40 ns and varied between
40 and 50 ns. The value dropped to 2.4 between 100 and 110 ns after reaching 3.1 between 80
and 90 ns. The MD simulation ended with it rising from 2.6 to 2.8. This suggests that the L1196
mutant's A-loop is more flexible than the normal type (see fig. 14).

(A) (B)
Figure 14. (A) Root Mean Square Deviation (RMSD) analysis of MD simulation trajectories
for (i) wild (4cmu) Ledipasvir-ALK TK, (ii) mutant (green, yellow) (4clj) Ledipasvir-ALK
TK, (iii) wild (4cmu) Ceritinib-ALK TK, and (iv) mutant (red, 4clj) Ceritinib-ALK TK. (B)
RMSD plot for the Apo-4cmu (non-complex) ALK TK protein.

Figure 19 demonstrates that the wild type A-loop RMSD converged to 1.8 after 40 ns of
simulation. The A-loop RMSD of the L1196 mutant increased to 2.4 at 40 ns and ranged from
40 to 50 ns. Between 80 and 90 ns, it hit 3.1 before dropping to 2.4 between 100 and 110 ns.
Later in the MD simulation, it rose from 2.6 to 2.8. The L1196 mutant's A-loop is more flexible
than the normal kind.

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Figure 15. Radius of gyration (Rg) trajectory study using MD simulations for [i] wild (4cmu)
and [ii] mutant (4clj) Ledipasvir-ALK TK, [iii] wild (4cmu) and [iv] mutant (4clj) Ceritinib-
ALK TK.

In figure 15, the C-backbone of ALK TK wild-type coupled to Ledipasvir has the least
compactness because its Rg plot fluctuates from 20.1 to 20.7, with a mean of 20,3 over 200 ns.
However, the C backbone of the ALK-TK mutant bound to Ledipasvir gyrated at 8.5 with
minor modifications. A reduction in Rg indicates a well-bound protein-ligand combination.
Mutant ALK-TK complexes with Ledipasvir were substantially more stable than wild-type
ones. Throughout the simulation, the RMSF plot showed each amino acid residue in its most
stable configuration. The amino acid residues in the ledipasvir-bound wild-type and mutant
ALK TK complex were least likely to alter (see fig. 16).

(A) (B)
Figure 16. (A) Root Mean Square Fluctuations (RMSF) in MD simulations of [i] wild (4cmu)
and [ii] mutant (4clj) Ledipasvir-ALK TK, [iii] wild (4cmu) and [iv] mutant (4clj) Ceritinib-
ALK TK. (B) Root Mean Square Fluctuations (RMSF) in MD simulations of Apo-ALK TK
(non-complexed ALK TK).

Global quality study of RMSD and Rg demonstrates that ledipasvir significantly affects the
stability of wild-type and mutant ALK TK targets in the binding cavities. Root mean square
fluctuation (RMSF) plots with a time function of 200 nanoseconds indicated considerable
RMSF at particular residues in both wild-type and mutant ALK-TK proteins. In contrast, apo-
ALK TK fluctuated less.

3.7 Molecular Mechanics Generalised Born Surface Area (MMGBSA)

MMGBSA is mostly used to measure ligand-protein binding. Ledipasvir-ALK TK wild-type
(4cmu) and mutant (4clj) complexes' binding free energies, ceritinib-ALK TK complexes'
binding free energies, and other non-bonded interaction energies were assessed. The Ledipasvir
ligand bound to the wild-type (4cmu) and mutant (4clj) ALK TK complexes at -47.77 and -
61.68 kcal/mol, respectively. Table 5 shows the average binding energies of the wild-type
(4cmu) and mutant (4clj) ceritinib-ALK TK complexes, -58.49 and -51.31 kcal/mol. Unlike

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ceritinib, ledipasvir bound to mutant ALK TK more strongly than native ALK TK. Gbind is
governed by non-covalent interactions such GbindCoulomb, GbindCovalent, GbindHbond,
GbindLipo, GbindSolvGB, and GbindvdW. GbindLipo and GbindCoulomb contributed less
energy than GbindvdW, but all three energies affected the binding affinity between wild-type
and mutant ALK TK and ceritinib. A key role for GbindvdW in drug receptor interactions has
been discovered. In contrast, GbindSolvGB and Gbind covalent energies had the least effect
on mean binding energies (see table 4).

Table 4 Presentation of the MMGBSA results for the ledipasvir and ceritinib wild and mutant
ALK TK strains.
Energies (kcal/mol) Ledipasvir- Ledipasvir- Ceritinib- Ceritinib-ALK
* ALK-TK wild ALK-TK ALK-TK wild TK-mutant (4clj)
(4cmu) mutant(4clj) (4cmu)
ΔGbind -47.77 ± 6.95 -61.68±8.16 -58.49±4.42 -51.31±6.29
ΔGbindLipo -17.98 ± 1.95 -22.92±2.55 -19.20±1.46 -17.51±2.01
ΔGbindvdW -52.58 ± 6.71 -66.37±5.71 -54.49±3.75 -54.21±5.77
ΔGbindCoulomb -17.51± 15.17 9.10±13.51 10.96±10.63 11.57±3.17
ΔGbindHbond -0.85 ± 0.47 -0.93±0.53 -0.95±0.50 -0.60±0.35
ΔGbindSolvGB 9.98 ± 13.37 14.32±12.01 -1.40±9.36 5.82±3.12
ΔGbindCovalent 6.86 ± 3.59 5.59±2.66 6.59±3.12 3.64±2.17

The wild-type and mutant ledipasvir and ceritinib-ALK TK complexes formed stable hydrogen
bonds with amino acid residues. Their GbindHbond interaction values showed this. Both
GbindSolvGB and GbindCovalent showed negative energy contributions for each molecule,
indicating binding resistance. Ledipasvir and ceritinib have transformed from curved to straight
in the binding pockets of wild-type and mutant ALK-TK proteins (200 ns). Changes in
conformation improve contact between the binding pocket and residues, increasing stability
and binding energy.

3.8 MTT Assay (In-Vitro Cell line study)

Ledipasvir and posaconazole, our most active hits, were tested in vitro on A549 cell lines for
anticancer activity. Ledipasvir and posaconazole were selected for in vitro cell line research
because they are in or near the QSAR model's applicability region.
In-Vitro Evaluation of Anticancer activity

100
% Inhibition (Posaconazole)
80 % Inhibition (Ledipasvir)
% inhibition

% Inhibition (Ceritinib)
60

40

20

0
0 20 40 60 80 100
Drug Conc. (μM)

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Figure 17: Depiction of in-vitro evaluation of anticancer activity.

Our study of in vitro anticancer activity on the A549 lung cancer cell line found that our
acquired hit, Ledipasvir, had a slightly higher inhibition rate than the standard reference
molecule, Ceritinib (see figure 17). The findings match computer predictions, indicating that
the same mechanism that reduced A549 lung cancer cells may inhibit ALK TK. However, we
believe further enzyme tests are needed to provide greater information.

4 Conclusion
A six-descriptor QSAR model was created for 1328 drugs exhibiting TK-ALK inhibitory
activity (IC50). The robust and predictive QSAR model meets all threshold values, including
R2 = 0.79, Q2LOO = 0.78, Q2LMO = 0.78, R2ex = 0.77, CCCex = 0.87, etc. The model discovered
many hidden pharmacophoric features, including the sum of aromatic carbon partial charges,
sp3-hybridized carbons within 4 bonds of non-cyclic carbons, and planer nitrogen within 6
bonds of the ring carbon atom. The Quantitative Structure-action Relationship (QSAR)
research showed that the aromatic or ring carbon atom, ring nitrogen atom, or non-ring nitrogen
atom significantly affected ALK TK's inhibitory action. As with other TK ALK inhibitors like
crizotinib and brigatinib, X-ray-resolved structures confirmed the mix of revealed and hidden
structural features. Additionally, QSAR-based virtual screening and drug repositioning
identified ledipasvir as an FDA-approved compound with a clinical trial IC50 of 0.65 nM
(PIC50–9.18 M). Ledipasvir outperformed ceritinib in molecular docking, scoring -10.57
kcal/mol against the ALK TK wild strain (pdb-4cmu) and -8.5286303 against the mutant strain
(pdb-4clj, mutant). Ledipasvir has a greater binding energy than ceritinib (-38.64 kcal/mol
against natural ALK TK and -34.64 against mutant ALK TK) and other ALK TK inhibitors,
according to docking research. Ledipasvir had a high affinity because the ALK-TK mutant
strain established more hydrophobic contact with it than the natural strain. The MD simulation
and MMGBSA analysis showed that the drug-receptor complex was stable beyond 200 ns and
had high binding energy. The mutant ALK TK-ledipasvir combination was much more stable
than the natural complex. Thus, MD simulation trajectories and binding energy docking data
validated MM-GBSA predictions. Additionally, the MTT test showed that ledipasvir had
somewhat stronger anticancer activity against the lung cancer cell line A549 than ceritinib. The
in-silico investigation confirms the in vitro anticancer effectiveness. These results may help
design a novel ALK-TK chemotherapy treatment.

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40
Synthesis, characterization and biocidal study of some novel 2-hydroxy 4,6-
disubstituted phenyl pyrimidines bearing 4-bromo phenol moiety

Rajendra M. Pathade*, Pravin. S. Bodkhe

Department of chemistry, Vidyabharati Mahavidyalaya, Amravati. * Email: [email protected]

ABSTRACT
In the present research work a series of 2-Hydroxy 4,6-disubstituted phenyl Pyrimidine
derivatives was synthesized from 4-bromo phenol as a precursor. The compound 4-bromo
phenol react with acetic anhydride to obtained 4-bromo phenyl acetate (1) then it follows Fries’
rearrangement reaction to form 5-bromo,2-hydroxy acetophenone(2).The compound(2)
allowed to react with different benzoic acids to give corresponding substituted 2-benzoyloxy
acetophenone (3a-d) which follow B.V.T. rearrangement reaction to obtained propan1,3-
diones(4a-d) and then it reacts with Urea to yield 2-hydroxy 4,6-disubstituted phenyl pyrimidine
derivatives (5a-d).The newly synthesized compounds are characterized by IR, H1NMR, Mass
spectral and elemental analysis. These compounds were also screened for their biocidal study
i.e., antibacterial, antifungal, anti-oxidant and anti-inflammatory activities.

KEYWORDS: 4-bromo phenol, Urea,2-Hydroxy 4,6-disubstituted phenyl Pyrimidine,

antibacterial, antifungal, anti-oxidant and anti-inflammatory activities.

INTRODUCTION
Heterocyclic compounds are abundant in nature and it have great significance to life. The
natural heterocyclic compounds such as vitamins, hormones, antibiotics etc. These heterocyclic
compounds containing nitrogen play an important role in medicinal chemistry and also
contribute to the society. Pyrimidine is a six-member heterocyclic compound which contains
two nitrogen atoms at positions 1 and 3. Pyrimidine derivatives are known to be biologically
active compounds and substituted pyrimidines have shown wide range of biological activities
like Anticancer [1,2], antibacterial [3-6], antifungal [7], antioxidant [8,9], anti-inflammatory
[10,11],anti-HIV [12],antitumor [13], antiparasitic [14],anti-Alzheimer's agent [15], anti-
diabetic agent [16].In addition to the diverse biological activities of pyrimidines play an
essential role in several biological processes and have a considerable chemical and
pharmacological importance. In the present research work some novel 2-hydroxy 4,6-
disubstituted phenyl pyrimidine(5a-d) compounds have been synthesized and studies their
structural characterization, antibacterial, antifungal, antioxidant and anti-inflammatory
activities.
EXPERIMENTAL SECTION
Materials and Methods
All the solvents and chemicals were of research grade and highest purity. The IR spectrum was
recorded by using Shimadzu IR affinity-1FTIR instrument, H1NMR spectra were recorded on
Bruker advance II 400 MHz spectrometer, Mass spectra were recorded on ESI and Melting
point were determined by open capillary tube method which are uncorrected.All the
synthesized compounds were purified by recrystallization and purity of the compound was
checked by TLC and elemental analysis.
General procedure for the Synthesis of 2-hydroxy 4,6-disubstituted phenyl Pyrimidine
derivatives (5a-d)
The synthesis involves the following steps.
Synthesis of 4-bromo phenyl acetate (1)

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Take 4-bromo phenol (0.05M) fused with acetic anhydride(5ml) and sodium acetate. The
mixture was refluxed for 1hr. then cooled for 15 min. and poured in ice water. Acetate layer
was separate out by separating funnel. The product was obtained 4-bromo phenyl acetate (1).
Synthesis of 5-bromo,2-hydroxy acetophenone (2)
Place aluminum chloride (120 g) in kjeldal flask and add compound (1)(40 ml) drop wise. Heat
the reaction mixture in oil bath for 60 min at 1200C. It was cooled and add in to acidified ice
crushed water to get crude product of 5-bromo,2-hydroxy acetophenone (2).
Synthesis of 2-substituted benzoyloxy 5-bromo acetophenone (3a-d)
A mixture of compound (2)(0.05M) and substituted benzoic acid (0.05M) were dissolved in
dry pyridine at 00 C.Then add POCl3 dropwise with constant stirring bellow 100C.The reaction
mixture was allowed to stand for overnight at room temperature.Then it was poured in ice cold
10% HCl.Then the product was wash by 10% NaHCO3 and water. Recrystallized the product
by ethanol to obtained a series of 2- substituted benzoyloxy 5-bromo acetophenone (3a-d).
Synthesis of substituted propane 1,3-diones (β-diketones) (4a-d)
Take compound (3a-d) (0.05M) was dissolved in dry pyridine. The reaction mixture was heated
up to 600C with pulverized KOH slowly with constant stirring. After 5-6 hr. the reaction
mixture was acidified by dil. HCl in ice cold water. The crude product was filtered, washed it
with NaHCO3 (10%) and water. Recrystallized the product from ethanol-acetic acid mixture to
get substituted propane 1,3-diones (4a-d).
Synthesis of 2-hydroxy-4,6-disubstituted phenyl pyrimidine derivatives (5a-d)
A mixture of compound (4a-d) (0.02 M) and urea (0.02 M) dissolved in DMF solvent. It was
refluxed on water bath at 750C for 1hr. then mixture was cooled and pour in ice cold water.
The product was recrystallized by aq. alcohol to obtained a series of 2-hydroxy-4,6-
disubstituted phenyl pyrimidine derivatives (5a-d)
General reactions scheme is given bellow.
OH
OH (CH CO) O O CH3 AlCl3
3 2 CH3
O Br
Br CH3COONa Br Fries Rearrangement O (2)
4-Bromo Phenol (1)
R1 Pyridine/
R2 COOH POCl3
Substituted Benzoic Acids
R1
O
OHR1 R2 O C R2
BVT Reaction
Br CH3
O O Pyridine / KOH Br
O
(4a-d)
(3-d)
Urea
R2
Where, 5a:- R1-H, R2 -CH3
OH 5b:- R1-H, R2-NO2
R1
Br N 5c:- R1-Cl, R2-H
N 5d:- R1-H, R2 -Cl
(5a-d) OH
Figure-1-The general reaction scheme for the synthesis of 2-hydroxy-4,6-disubstituted phenyl
pyrimidines (5a-d)

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Table No.1: Physical data of 2-hydroxy-4,6-disubstituted phenyl Pyrimidines (5a-d)

Code Compound Name M.F./ Rf M.P. Yield
value
M.W. (0C) (%)
5a 2-hydroxy-4-(4-methyl phenyl)-6-(2- C17H13BrN2O2 / 0.72 80 -
hydroxy-5 bromo phenyl) pyrimidine
(357.20) 86 58%
5b 2-hydroxy-4-(4-nitro phenyl)-6-(2- C16H10BrN3O4 / 0.68 116 -
hydroxy-5 bromo phenyl) pyrimidine 122
(388.17) 71%
5c 2-hydroxy-4-(2-chloro phenyl)-6-(2- C16H10BrClN2O/ 0.66 74 - 61%
hydroxy-5 bromo phenyl) pyrimidine
(377.96) 78
5d 2-hydroxy-4-(4-chloro phenyl)-6-(2- C16H10BrClN2O/ 0.70 112- 65%
hydroxy-5 bromo phenyl) pyrimidine 116
(377.96)
Antimicrobial Activity
The in-vitro anti-microbial screening of newly synthesized 2- hydroxy -4,6-disubstituted
phenyl pyrimidines (5a-d) was carried out against bacteria staphylococcus aureus (gram
+ve),Salmonella typhus (gram–ve) and fungi Candida albicans and aspergillus niger by disc
diffusion method[17,18] and compared with that of the standard drugs Oxacillin (2 µg) and
Fluconazole(25µg) respectively of each drug was defined as the lowest concentration of an
antimicrobial that will inhibit the visible growth of microorganism after incubation time.
Muller Hinton Agar was used as basal medium for test of bacteria and fungi respectively.
The compounds tested at concentration of 50 µg/ml, 100 µg/ml and 250 µg/ml for bacterial
and fungal growth in DMSO solvent it was added to the wells made on culture medium.
0
After 24 hrs. of incubation at 37 C for antibacterial activity and after 24 hrs. at room temp. for
antifungal activity, record the zone of inhibition was compared with the standard drug
Oxacillin and Fluconazole.
Anti-Inflammatory Activity
Take 10 mg of Ibuprofen as a reference drug was added to10 ml of distilled water. Serial
dilution from above stock solution takes 0.1ml, 0.2ml, 0.3ml and prepare 10 ppm, 20 ppm and
30 ppm and also it was performed for sample 2- hydroxy -4,6-disubstituted phenyl pyrimidines
(5a-d) extract. The reaction mixtures were prepared using 2.8 ml of phosphate-buffered saline
(pH 6.4) and 0.2 ml of egg albumin. Then take 2 ml of extract from each different concentration
were mixed gently with reaction mixtures. A similar procedure was used for reference drug
ibuprofen. The absorbance of these solutions was determined by using spectrophotometer at
wavelength of 660 nm. The % denaturation of the protein (% inhibition) was determined.[19]
Anti-Oxidant Activity
Stock solution of DPPH (2,2-diphenyl-1-picrylhydrazyl) was prepared by dissolving 1.083 mg
in 10 ml of ethanol. Stock solution of sample 2-hydroxy-4,6-disubstituted phenyl pyrimidines
(5a-d) take 100 μg/ml was prepared by dissolving 1 ml of sample in 10 ml of ethanol. From
this stock solution, further dilutions were prepared of concentrations 10,20,30,40 and 50 μg/ml
using ethanol.Similarly,stock solution of standard ascorbic acid was prepared by dissolving 10
mg ascorbic acid in 10 ml ethanol.From this stock solution further dilutions of concentrations
1,2,3,4 and 5μg/ml were prepared.Absorbance of blank(5 ml ethanol+1 ml DPPH solution) as
a positive control was recorded using colorimeter at 517 nm.Similarly,the absorbance of

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sample and comparative standard ascorbic acid was taken at 517 nm.The % scavenging activity
was determined.[20]
RESULTS AND DISCUSSION
Spectroscopic characterization data
(5a) 2-hydroxy-4-(4-methyl phenyl)-6-(2-hydroxy-5 bromo phenyl) pyrimidine
Solid,yellow,IR(cm-1):3200(Ar-OH),3058(Ar-C-H),2928(Ar-CH3),1613(C=N),1458(C=C),
640 (C-Br),1280(C-O);1H-NMR(δppm):δ7.90-7.98(m,8H of Ar-H),δ7.36-7.40 (s,1H of -
OH),δ 2.38 (s,3H of -CH3);MASS (m/z,%):358(M+); C,H,N,O% Calculated(Found) C:57.16
(56.86),H:3.67 (3.61),N:7.84 (7.62),O:8.96 (8.17).
(5b) 2-hydroxy-4-(4-nitro phenyl)-6-(2-hydroxy-5 bromo phenyl) pyrimidine
Solid, yellow, IR (cm-1):3379 (Ar-OH),3096 (Ar-C-H),1605 (C=N),1456 (C=C),705 (C-
Br),
1207 (C-O),1345 (-NO2);1H-NMR (δ ppm):δ7.2-8.4 (m,8H of Ar-H),δ 6.85 (s,1H of Ar-
OH); MASS (m/z,%):388.99 (M+);C,H,N,O% Calculated (Found):C:49.51 (48.29),H:2.60
(2.13), N:10.83 (10.01),O:16.49 (15.59).

(5c) 2-hydroxy-4-(2-chloro phenyl)-6-(2-hydroxy-5 bromo phenyl) pyrimidine

Solid, brown, IR (cm-1):3076 (Ar-OH),2927 (Ar-C-H),1637 (C=N),1459 (C=C),529 (C-Br),
1275 (C-O),757 (C-Cl);1H-NMR (δ ppm): δ7.31-8.19 (m,8H of Ar-H),δ 6.69 (s,1H of Ar-
OH);MASS (m/z,%):377.97(M+); C, H, N, O% Calculated (Found): C:50.89 (50.02),H:2.67
(2.13),N:7.42 (7.15),O:8.47 (7.98).
(5d) 2-hydroxy-4-(4-chloro phenyl)-6-(2-hydroxy-5 bromo phenyl) pyrimidine
Solid, yellow, IR (cm-1):3188 (Ar-OH),3088 (Ar-C-H),1610 (C=N),1465 (C=C),527 (C-Br),
1274 (C-O),641 (C-Cl);1H-NMR (δ ppm):δ7.90 -7.98 (m,8H of Ar-H),δ7.36 -7.40 (s,1H of
Ar-OH);MASS (m/z, %):379(M+);C,H,N,O % Calculated (Found): C:50.89 (50.12),H:2.67
(1.99),N:7.42 (6.76),O:8.47 (8.17).

BIOCIDAL STUDY
Table No.2- Antibacterial activity of 2-hydroxy 4,6-disubstituted pyrimidines (5a-5d)
Zone of inhibition in mm
Salmonella typhi (gram -tive) Staphylococcus aureus (gram +tive)
Compound Concentrations µgm/ml Concentrations µgm/ml
code Standard Standard
50 100 250 Ofloxacin 2 mcg 50 100 250 Ofloxacin 2 mcg
5a ++ ++ ++ ++ ++ +++ +++ +++
5b ++ ++ ++ - - - - -
5c - - - - ++ +++ +++ -
5d - - - - - ++ ++ -
Highly active+++=13-24,Moderate active++=7-12,Less active+=1-6,No zone of inhibition-

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Table No.3- Antifungal activity of 2-hydroxy 4,6-disubstituted pyrimidines (5a-5d)

Zone of inhibition in mm
Candida albicans Aspergillus niger
Compound Concentrations µgm/ml Concentrations µgm/ml
code Standard Standard
50 100 250 Fluconazole 25 µgm 50 100 250 Fluconazole 25 µgm
5a - - - +++ - - - +++
5b - - ++ - - ++ ++ -
5c - ++ +++ - - - ++ -
5d - - - - - - - -
Highly active+++=13-24/Moderate active++=7-12/Less active+= 1-6/No zone of
inhibition-
The antibacterial activity results revealed that compounds showed significant activity against
gram +ve organisms. The compound 5a and 5b showed moderate activity against Salmonella
typhi and compounds 5a and 5c shows highly activity and 5d shows moderate activity against
Staphylococcus aureus.The compounds 5c and 5d does not showed any zone of inhibition
against the gram –ve organism. In the antifungal activity, compound 5b and 5c showed moderate
to weak activity against Candida albicans and Aspergillus niger.The compounds 5a and 5d does
not showed any zone of inhibition in fungal activity at different concentration.

Table No.4-Anti-inflammatory activity of 2-hydroxy 4,6-disubstituted pyrimidines(5a-d)

% Inhibition
Sr. Compound Concentration of compound in ppm
No. Code 10 ppm 20 ppm 30 ppm
1 5a 40.67 63.68 73.06
2 5b 45.47 53.75 68.76
3 5c 6.34 17.88 59.71
4 5d 66.33 77.81 79.41
5 Ibuprofen 87.36 88.90 90.23

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Graph No.01-Anti-Inflammatory Activity

% of Inhibitions
100

50

0
10 ppm 20 ppm 30 ppm
Concentration 5a 5b 5c 5d Ibuprofen

Table No.5- Anti-oxidant activity of 2-hydroxy 4,6-disubstituted pyrimidines(5a-d)

% scavenging activity
Sr. No. Compound Code Concentration of compound in μg/ml
10 μg/ml 20 μg/ml 30 μg/ml 40 μg/ml 50 μg/ml
1 5a 68.43 69.08 78.95 81.58 98.46
2 5b 46.93 57.68 64.48 66.89 88.82
3 5c 63.16 64.04 64.48 78.51 94.08
4 5d 50.88 54.61 56.80 59.87 60.09
5 Ascorbic Acid (Standard) 80.26 91.22 95.61 96.27 97.80

Graph No.02-Antioxidant Activity

150
5a 5b 5c 5d Ascrobic Acid
% of scavenging

100

50

0
10 20 30 40 50
Concentratios µg/ml

CONCLUSION
In the present work newly synthesized 2-hydroxy 4,6-disubstituted phenyl Pyrimidines
containing 4-bromo phenol moiety involves different steps to get good yield. The structure of
synthesized compound was elucidated on the basis of 1H-NMR, IR and Mass spectra. In
antibacterial activity compound 5a and 5b shows moderate activity against Salmonella typhi
and the compound 5a,5c and 5d shows highly active to moderate zone of inhibition against
Staphylococcus aureus. In antifungal activity compound 5b and 5c showed moderate to weak
activity against Candida albicans and Aspergillus niger at concentration 100 µgm and 250 µgm.
Also compounds 5a,5b,5c and 5d exhibited significant anti-inflammatory activity by using
albumin denaturation technique at 30 ppm concentration. Antioxidant activity as indicated by
absorbance of compounds 5a-5d increased with increasing concentration. Higher value of
absorbance of the reaction mixture indicated greater reducing power. The reducing power was
found to be in order of 5a>5c>5b>5d

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ACKNOWLEDGEMENT
The author is thankful to the Principal, Vidyabharati Mahavidyalaya, Amravati for providing
the laboratory facilities and Director, SAIF Panjab University, Chandigarh for providing NMR,
IR and Mass spectra analysis. Also thankful to Prof.Dr. Sharda Deore Government Pharmacy
college, Amravati for Pharmacological activity and Dr.S.R. Gulhane, Microbiology Diagnostic
Lab, Amravati for biocidal activity.
REFERENCES
[01] Zhao A.,Gao X.,Wang Y.,Ali J.,Wang Y.,Chen Y. et al. ‘Discovery of novel c-Met kinase
inhibitors bearing a thieno [2,3-d] pyrimidine or Furo[2,3-d] pyrimidine scaffold’ Bioorgan Med
Chem,2011,19:3906-3918.
[02] Shaaban MA, Mohamed KO, Hegazi ME, Montaser A. Shaykoon, Yaseen AMM Elshaier
‘Synthesis of Some Pyrimidine and Fused Pyrimidine Derivatives with Antimicrobial and Anticancer
Activities’ Der Pharma Chemica, 2018;10(7):180-200.
[03] Liu, Yu; Sun, Xiao; Yin; Da; Yuan, Fang, ‘Syntheses and biological activity of chalcones
-imidazole derivatives’Research on Chemical Intermediates,2013, Vol.39, pp.1037-1048.
[04] Abdel Gawad MA. Synthesis and antibacterial evaluation of new azo-pyrimidine derivatives. J
Appl Pharm Sci. 2019; 9(1):9-16
[05] Sebastin V. et al, “Synthesis and Antibacterial Activity of Pyrimidine Derivatives of 1,3-
Dihydropyrimidine’’Am. J Pharm Tech Res.2017;7(5):24-29
[06] Binani et al.“synthesis, characterization and in vitro antimicrobial evaluation of novel 2-
mercapto-4,6-disubstituted phenyl pyrimidine derivatives” int j pharm pharm sci, vol 6, issue 1,2014,
461-463, ISSN- 0975-1491.
[07] Y. M. Zohny et al./Pharmacophore 2015, Vol. 6 (6), 255-266.
[08] K.Elumalai,M.A.Ali, M.Elumalai, K. Eluri, S. Srinivasan, J Acute Disease,2013, 316-321.
[09] T.N. Doan, D.T. Tran. Pharmacol. Pharm.,2011, 2, 282-288.
[10] C.M. Bhalgat, M.I. Ali, B. Ramesh, G. Ramu, Ara J Chem.,2011, 1-8.
[11] Bhalgat CM.,et al.“Novel pyrimidine and its triazole fused derivatives:Synthesis and
investigation of antioxidant and anti-inflammatory activity”Arabian Journal of Chemistry 7.6, 2014,
986-993.
[12] Kashyap,S.J. et.al J.Adv.Sci.Res.2011,2, 18-24.
[13] Li Q.,et al.“Synthesis and biological activity of fused furo [2,3-d] pyrimidinone derivatives as
analgesic and antitumor agents”.Research on Chemical Intermediates 42.2 , 2016, 939-949.
[14] Azas, N.; Rathelot, P.; Djekou, S.; Delmas, F.; Gellis, A.; Giorgio, C. D.;Vanelle,
P.;Timon-David,P.Farmaco 2003,58,1263-1270.
[15] Elmegeed G.A., Ahmed H.H., Hashash M.A., Abd-Elhalim M.M., El-kady D.S. “Synthesis of
novel steroidal curcumin derivatives as anti-Alzheimer’s disease candidates: evidences-based on in
vivo study Steroids”. 2015, 101:78–89.
[16] Barakat A. et al.“Synthesis and structure investigation of novel pyrimidine-2,4,6-trione
derivatives of highly potential biological activity as anti-diabetic agent”, Russian Journal of
Bioorganic Chemistry 41, 2015,192-200.
[17] Sambhaji P. Vartale et al. ‘Synthesis and antimicrobial evaluation of pyrimido pyrimidine
derivatives’, IJRPC 2015,5(1),208-214 ISSN:2231-2781.
[18]Vijay V.Dabholkar and Ashish S.Sanghvi.Indian Journal of Heterocyclic Chemistry, 2006,
16:105.
[19]Monica Kachroo et. al.“Synthesis and Biological Activities of Some New Pyrimidine
Derivatives From Chalcones’’Der Pharma Chemica, 2014,6(2),352-359.
[20]Reşat Apak et al. J. Agric. Food Chem.2016,64,997−1027,DOI: 10.1021/acs.jafc.5b04739.

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41

Investigation of antimicrobial Activities of Dihydropyrazines

Manish Fukatea, Sheryil Malvilayilsa,Roshan Jaiswalb,Swati Bawiskarb
a G H Raisoni University Amravati a G H Raisoni University Amravati
b G H Raisoni University Saikheda b G H Raisoni University Saikheda
corresponding author [email protected]

Abstract
Seven Dihydropyrazines 4ag are screened for their antibacterial activity against
Staphylococcus aureus, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa,
Salmonella typhi and antifungal activity against Candida albicans, Aspergillus flavus, Rhizopus
and Mucor. Ciprofloxacin is used for the standard for antibacterial and Amphotericin B is used
for the standard for antifungal studies. Compounds 4b, 4c,4fand 4gexhibited excellent in vitro
antibacterial activity against all the tested organisms. Whereas the same set of compounds
exerted potent in vitro antifungal activity against Candida albicans, Aspergillus flavus and
Rhizopus.
Keywords: Dihydropyrazines,Antibacterial activity, antifungal activity, Ciprofloxacin,
Amphotericin B.
1. INTRODUCTION
The biological and physical roles of dihydropyrazines(DHPs)such asDNA
cleavage[1],growthinhibitionofEscherichiacoli[2],Cyclooxygenaseinhibitoryactivity [3] and
NPY antagonists [4] are well documented. Dihydropyrazinesare universal in the human
body[5]however, there is little reported concerning the biological and physiological roles of
DHPs.Yamaguchi etal.[1]reported generation of free radicals from
dihydropyrazineswithDNAStrand-
Breakageactivity.Takechietal.[6]reportedthegrowthinhibitionandmutagenesisinducedin
Escherichia coli by dihydropyrazines with DNA strand-cleavingactivity.2-
cyanopyrazinederivativesshowanticancerantiinflammatoryandanalgesicactivities[7].Pyrazine
derivatives exits a tuberculostatic activity [8]. It also exhit a antimicrobial [9] and biological
[10] activities. Alkyl substituted pyrazines are found in the growth medium of the polymyxin-
producingbacterium Paenibacilluspolymyxa[11].
The se observations place snew emphasis on then need of as well as search for
alternative new and more effective antimicrobial agents with abroad spectrum.
The aim of this study was to evaluate the biological activities of dihydropyrazines
4a4g. The results of the antibacterial and antifungal activities are discussed in this paper. To
percept structure-activity relationship well, numberings of the target compound is shown
below Fig. 1.

Fig.1Numberingof 4a-g

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2. MATERIALS & METHODS

2.1 Materials
All the bacterial strains namely Staphylococcus aureus, Escherichia coli, Klebsiella
pneumoniae, Pseudomonas aeruginosa, Salmonella typhi and fungal strains namely
Candida albicans, Aspergillus flavus, Rhizopus and Mucor were obtained
2.2 Methods
The compounds were synthesized by the following literature procedure .12 The in vitro
antimicrobial activities of the compounds were tested in Sabouraud’s dextrose broth (SDB,
Hi-media, Mumbai) for fungi and nutrient broth (NB, Hi-media, Mumbai) for bacteria by the
twofold serial dilution method.13
The test compounds were dissolved in dimethyl sulfoxide (DMSO) to obtain 1 mg/ml
stock solutions. Seeded broth (broth containing microbial spores) was prepared in NB from 24
h old bacterial cultures on nutrient agar (Hi-media, Mumbai) at 37  1 C while fungal spores
from 24 h to 7-day-old Sabouraud’s agar slant cultures were suspended in SDB. The colony
forming units (cfu) of the seeded broth were determined by the plating technique and adjusted
in the range of 104105 cfu/ml. The final inoculum size was 10 5 cfu/ml for the antibacterial
assay and 1.11.5  102 cfu/ml for the antifungal assay. Testing was performed at 7.4  0.2.
Exactly 0.2 ml of the solution of test compound was added to 1.8 ml of seeded broth to form
the first dilution. One ml of this was diluted with a further 1 ml of the seeded broth to give the
second dilution and so on until six such dilutions were obtained. A set of assay tubes containing
only seeded broth was kept as control and likewise solvent controls were also run
simultaneously. The tubes were incubated in biochemical oxygen demand (BOD) incubators
at 37  1 C for bacteria and 28  1 C for fungi. The minimum inhibitory concentrations
(MICs) were recorded by visual observations after 24 h (for bacteria) and 7296 h (for fungi)
of incubation. Ciprofloxacin was used as a standard for the bacterial study while Amphotericin
B was used as a standard for the fungal study.
3. RESULT & DISCUSSION
Physical data of the compounds 4a-4g are given in the Table 1.

Table 1. Physical data of the compounds 4a-4g

Compound R Yield (%) Melting Point ◦C
4a H 96 242- 243
4b 4-Cl 98 211 - 212
4c 2-Cl 92 235- 236
4d 4-CH3 96 254- 255
4e 4-OCH3 98 208- 209
4f 4-NO2 94 284 - 285
4g 4-F 96 236 - 237

In vitro antibacterial and antifungal activity

Compounds 4a4g were tested for their antibacterial activity in vitro against
Staphylococcus aureus(SA) , Escherichai coli (EC), Klebsiella pneumonia(KP), Pseudomonas
aeruginosa(PA) and Salmonella typhi (ST). Ciprofloxacin was used as standard drug.
Minimum inhibitory concentration (MIC) in μg/ml values shown in Table 2.

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Table 2. In vitro antibacterial activity of compounds 4ag

Minimum Inhibitory Concentration (MIC) in μg/ml

Compound SA EC KP PA SA
4a 100 200 200 200 100
4b 25 25 50 25 25
4c 25 25 50 25 25
4d 50 100 100 100 50
4e 50 100 100 50 50
4f 6.25 6.25 6.25 12.5 6.25
4g 3.13 3.13 6.25 3.13 3.13
Ciprofloxacin 25 25 12.5 25 25

All the 3,4-dihydropyrimidin-2(1H)-ones 4a4g exerted potent antibacterial activity in

vitro against the tested bacterial strains. Moreover, compounds 4b, 4c, 4fand 4g exerted
excellent antibacterial activities aga inst S. aureus, E. coli, K. pneumoniae, P. aeruginosa and S.
typhi. A comparative study of minimum inhibitory concentration for the compounds 4a4g
using standard Ciprofloxacin versus bacterial strains given in Fig. 2.

250

200

150
SA

100 EC
KP
50 PA
ST
0

Fig. 2. Comparison of minimum inhibitory concentration of compounds 4ag

with Ciprofloxacin (as standard) against bacterial strains from serial dilution method.

The in vitro antifungal activity of the synthesized compounds 4a4g was studied
against the fungal strains viz., Candida albicans (CA), Aspergillus flavus (AF), Rhizopus and
Mucor. Amphotericin B was used as a standard drug. Minimum inhibitory concentration (MIC)
in μg/ml values is shown in Table 3.

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Table 3. In vitro antifungal activity of compounds 4ag

Minimum Inhibitory Concentration (MIC) in

Compound μg/ml
CA AF Rhizopus Mucor
4a 100 200 200 200
4b 25 50 25 25
4c 25 50 25 25

4d 50 100 50 50
4e 50 100 100 50
4f 6.25 6.25 12.5 6.25
4g 3.13 3.13 6.25 3.13
Amphotericin 25 25 50 25
B

Compounds 4b, 4c, 4f and 4g exhibited excellent antifungal activities against all the tested
fungal strains except Mucor. Minimum inhibitory concentration of compounds 4a4g were
compared with standard Amphotericin B against fungal strains shown in Fig 3.

250

200 CA
AF
150 R
M
100

50

Fig. 3. Comparison of minimum inhibitory concentration of compounds 4ag with

Amphotericin B (as standard) against fungal strains from serial dilution method.
4. Conclution
Results of this study show that compounds 4b, 4c 4f and 4g which contain chloro, nitro
and fluoro moieties exerted excellent antimicrobial activities against the tested organisms.
Further development of this group of 3,4-dihydropyrimidinones may lead to compounds with
better pharmacological profile than standard drugs and serve as templates for the construction
of better drugs to come to blows bacterial and fungal infections.

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References

[1] Yamaguchi T, Matsumoto S, Watanabe K. Generation of free radicals from

dihydropyrazines with DNA strand-breakage activity. Tetrahedron Lett. 1998; 39: 8311
- 8312.
[2] Takeda O, Takechi S, Katoh T, Yamaguchi T .The role of dihydropyrazines in
accelerated death of Escherichia coli on addition of copper(II). Biol Pharm Bull. 2005;
2 8 : 1161 - 1164.
[3] Singh S K, Saibaba V, Ravikumar V, Rudrawar S V, Daga P, Rao C S, Akhila V, Hegde
P, Rao Y K. Synthesis and biological evaluation of 2,3-dihydropyrazines and
quinoxalines as selective COX-2 inhibitors. Bioorg Med Chem . 2004; 12: 1881 - 1893.
[4] Sit S Y, Huang Y, Antal-Zimanyi I, Ward S, Poindexter G S. Novel dihydropyrazine
analogues as NPY antagonists. Bioorg Med Chem. 2007; 12: 337 – 340.
[5] Yamaguchi T, Nomura H, Matsunaga K, Ito S, Takata J, Karube Y. The behavior of
dihydropyrazine with DNA strand-breakage activity in vivo. Biol Pharm Bull. 2003;26:
1523 - 1527.
[6] Takechi S, Yamaguchi T, Nomura H, Minematsu T, Nakayama T. Growth inhibition and
mutagenesis induced in Escherichia coli by dihydropyrazines with DNA strand-cleaving
activity. Mutation Res. 2004; 560: 49 - 55.
[7] Sondhi S M, Singh N, Rajvanshi S, Johar M. Synthesis, hydrolysis over silica column,
anticancer, anti-inflammatory and analgesic activity evaluation of some pyridine and
pyrazine derivatives. Indian J Chem. 2005; 44B: 387 – 399.
[8] Foks H, Trapkowska I, Janowiec M, Zwolska Z, Augustynowicz-Kopec E. Synthesis,
reactions and tuberculostatic activity of pyrazinyl-substituted derivatives of
hydrazinocarbodithioic acid. Chem Heterocyclic Compd. 2000; 40: 1185 – 1193.
[9] Farghaly A R, El-Kashef H. Pyrazoles and pyrazolo [4,3-e] pyrrolo [1,2-a] pyrazines,
I. Synthesis and antimicrobial activity. Monatshefte Fiir Chemie. 2005; 136: 217 - 227.
[10] El-Kashef H S, El-Emary T I, Gasquet M, Timon-David P, Maldonado J, Vanelle P.
New pyrazolo [3,4-b] pyrazines: synthesis and biological activity. Pharmazie. , 2000;
55: 572 - 576.
[11] Beck H C, Hansen A M, Lauritsen F R. Novel pyrazine metabolites found in polymyxin
biosynthesis by Paenibacillus polymyxa. FEMS Microbiol Lett. 2006; 220: 67 – 73.
[12] Baliah V, Lakshmanan M R, Pandiarajan K. Synthesis of some 1,2-ethanediamines.
Indian J Chem, Vol. 1990; 16B: 72-73.
[13] Dhar M H, Dhar M M, Dhawan B N, Mehrotra B N. Screening of Indian plants biological
activity Part I. 1968; 6: 232-247.

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42
Synthesis and Spectral Characterization of Novel 1,2,4-Dithiazole
Derivatives
Mr Ramdas N Ingole1, Dr Siddharth S Waghmare1 and Dr Pravin S Bodkhe2
1
Research Center, Department of Chemistry, Ghulam Nabi Azad Arts, Commerce and Science College
Barshitakli District Akola 444401 (M.S.), India.
2
Research Center, Department of Chemistry, Vidya Bharti Mahavidyalaya, Amaravti.
e-mail:[email protected]
Abstract
A single step synthesis of N-(5-(substitutedimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-
amine (II a-d) was carried out by oxidative cyclisation of 2-(5-substituted-2,4-dithiobiureto)
benzothiazole (I a-d) using liquid bromine in chloroform medium as an oxidizing agent. The
products were isolated, characterized and justified on the basis of conventional chemical
characteristics and spectral studies.
Key-words: Benzothiazole, oxidative cyclization, dithiazoles, oxidizing agent.
Introduction
Organic compounds containing thiadiazole as the heterocycles in their structure are identified
for their biological activities, pharmaceutical activities, industrial applications, agricultural
purposes and in medicinal sciences. Basically structure of thiadiazole is five membered
nitrogen and sulphur containing heterocyclic compound. Presence of sulphur and nitrogen in
its structure enhances the biological potential of the thiadiazole. Recently reported some newer
thiadiazole along with pyridine for the anti-bacterial activities1-2 and effect of germination
pattern of jowar, Bhagwatkar3 synthesized and reported the thiadiazole based triazine for the
antibacterial activities and some different substituent’s attached thiadiazole directly or
indirectly shows effect directly in their biological, pharmaceutical and agricultural
applications4-5. The benzothiazole based heterocycles are five membered heterocycles
containing sulphur and nitrogen as a heteroatom were created its own background in the
synthetic organic chemistry6-10. The biological as well as industrial applications of
benzothiazole get enhanced, due the presence of the thiadiazole nucleus in their structure.
Trusting the literature ideas in mind it is decided to design such organic moiety should contain
thiadiazole along with the benzothiazole; to synthesize such series of benzothiazole based five
member heterocycle containing sulphur and nitrogen as a heteroatom11-17.
As a part of research work presently it has been planned to design and synthesize novel series
of N-(5-(substitutedimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-amine (II a-d) in this
laboratory with the easiest and cheaper method by oxidative cyclisation of 2-(5-substituted-
2,4-dithiobiureto) benzothiazole (I a-d) with liquid bromine in chloroform medium. The
present method utilized somewhat suitable, convenient, cheaper, more practical utility and only
a single step direct method for the synthesis of (IIa-d).
Experimental Section
Materials:
All chemicals used were of Mercks Millipore (Indian made). 2-(5-substituted-2,4-
dithiobiureto) benzothiazole (I a-d) were prepared by known literature method3 .
Method:
Method employed in the present experiments for the synthesis of series 1,2,4-dithiazole based
benzothiazole is conventional refluxing under water bath for different hours for different
experiments. The melting points of synthesized compounds were recorded using hot paraffin

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bath and uncorrected. IR spectra were recorded on Perkin Elmer spectrometer in the range
4000-400 cm-1 in KBr pellets. PMR spectra were recorded on BRUKER AIIIANCE II 400
NMR spectrometer with TMS as an internal standard using CDCl3 and DMSO-d6 as a solvent.
The purity of the compounds was checked on silica gel – G plates by TLC with layer thickness
of 3mm.
General Procedure:
Experiment No. 1
Synthesis of N-(5-(allylimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-amine (IIa):
The pest of 2-(5-allyl-2,4-dithiobiureto) benzothiazole (Ia) was prepared in chloroform in a
clean china dish and liquid bromine in chloroform was added with constant stirring at room
temperature. During the addition of bromine in chloroform solution to the pest of (Ia), firstly
the colour of bromine disappear, further addition of bromine in chloroform colour of bromine
appears and persists. Such solution of bromine in chloroform and (Ia) kept for 4 hours at room
temperature. Basification of the reaction mixture with dilute ammonium hydroxide solution
gives the formation bright cream yellow coloured product (IIa). Recrystallization of the product
was done using ethanol. Yield 89%, M. P. 1720C.
Similarly, N-(5-(ethylimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-amine (IIb), N-(5-
(phenylimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-amine(IIc), N-(5-(t-butylimino)-5H-
1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-amine (IId) were synthesized from the oxidative
cyclisation of 2-(5-ethyl-2,4-dithiobiureto) benzothiazole (Ib), 2-(5-phenyl-2,4-dithiobiureto)
benzothiazole (Ic), 2-(5-t-butyl-2,4-dithiobiureto) benzothiazole (Id) with liquid bromine in
chloroform medium respectively by the above mentioned method in Experiment No. 1 to 4 and
the data obtained by the characterization of synthesized compound in a series (IIa-d) is given
result section.
Reaction Scheme:
S S

NH N NHR
H
N

2-(5-substituted-2,4-dithiobiureto) benzothiazole (I a-d)

[O] Br2 / CHCl3

S S
S
N R
N
H N
N

N-(5-(substitutedimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-
amine (II a-d)

Where R = -allyl, -ethyl, -phenyl, t-butyl

Results and Discussion:

Spectral data obtained from the present research support the formation of designed or target
products. Spectral characterizations of all the synthesized compounds are also given below:
Data Analysis:

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N-(5-(allylimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-amine (IIa):
Cream Yellow solid, C13H11N3S3 , Yield-89 %, M.P. 272, FTIR (KBr) ν cm-3085.89-3004.89
(Ar C-H stretching), 574.75(S-S stretching), 1587.31 (S-C=N stretching ), 794.62-761.83 (C-
S stretching); 1H NMR (400 MHz CDCl3 δ ppm), doublet of 2H of –CH2 at δ 2.00 ppm,
singlet of 1H of NH at δ 4.00ppm, pentate of 1H, doublet 2H and doublet of 2H of allyl at
δ5.70ppm, 4.97ppm and 5.0ppm respectively and multiplet of 7H of Ph at δ 7.55-8.23ppm;
Mol. Wt.:305.
N-(5-(ethylimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-amine (IIb)
Dark Yellow solid, C11H10N4S3 , Yield-92%, M.P. 1680C, FTIR (KBr) ν cm-3085.89-
3004.89(Ar-H Stretching), 574.75(S-S stretching), 1587.31(-C=N stretching), 794.62-
761.83(C-S stretching),; 1H NMR (400 MHz CDCl3 δ ppm), triplet of 3H, -CH3 at
δ0.90ppm, quartrate of 2H of –CH2 at δ 1.4ppm, multiplet of 7H, Ph at δ 6.52-8.01ppm, singlet
of 1H, NH at δ 4.35ppm ; Mol. Wt.: 294.
N-(5-(phenylimino)-5H-1,2,4-dithiazol-3-yl)benzo[d]thiazol-2-amine(IIc)
Yellow, C15H10N4S3, Yield-88%, M.P. 174oC, FTIR (KBr) ν cm- 3056.96-3004.89 (Ar-H
Stretching), 592.11 (S-S stretching), 1587.31 (-C=N stretching), 794.62-761.83 (C-S
stretching),; 1H NMR (400 MHz CDCl3 δ ppm), multiplet of 12H, Ph at δ 6.11-8.02ppm and
singlet of 1H, -NH at δ4.34ppm; ESI-MS (m/z) gives base at m/z+=226.98, 74.90, 359.93 and
molecular ion i.e. molecular weight is 342.
In the present work is best cheaper and less time consuming method of cyclisation of organic
compound. Route of mechanism of the present synthesis is due to the conjugation of amido-H
with thionyl Sulphur. Among all the synthesised of compounds (IIa-IId), percentage of yield
of compound (IIb) is highest i.e. 92%.. However (IIa) and (IId) shows comparable yield with
the (IIb).
Conclusion
In all the synthesized compounds, maximum yield of (IIb) displays ethyl group is
electrometrically donating group on nitrogen in 2,4-dithiobiurets with increases the rate of
reaction by giving its electrons.

References
[1] PP Pathe, MW Ambekar, NM Nimdeokar, MG Paranjpe; Indian J. Chem., 1982, 59, 670.
[2] AK Bhagwatkar; Synthesis and antimicrobial analysis of substituted 1,3,5-triazin-6-
ylsubstitutedamidinothiocarbamides and their cyclisation into 1,3,5-thiadiazines and 1,3,5-triazines’, Ph.D.
Thesis, SGBAU, Amravati, 2013.
[3] AKSingh, G Mishra and K Jyoti; Journal of App. Pharm. Science, 2011, 1(5), 44-49.
[4]J Tao, ZW Duo and HC Ling; J Chinese Chem Soc., 2010, 57,1077-80.
[5] NV Madhav, AS Nayak, AJ Rao, and M Sarangapani; J Pharm Res., 2011, 4(5), 1396-97.
[6] D Patel and A Singh; -Jour Chem., 2009, 6(4), 1017-1022.
[7] PR Mahalle and SP Deshmukh; Int. J. of Pharmcy and Pharmcaeutical Science, 2011, 3(4), 277-279.
[8] MA Saleh, Sulfur letters, 2002, 25, 235‐245.
[9] F Sansone, E Chierci, A Casanti and R Ungaro; Org. Bio. Mol. Chem, 20031, 1802‐1803.
[10]PR Mahalle and SP Deshmukh, Int. J. of Carbohydrate Research, 2012, 1(1), 1-3.
[11]KM Heda and SP Deshmukh, Rasayan J. Chem., 2012, 5(1), 24-27.
[12]K Willi, B Helmut, K Wolfgang, M Edger and R Peter; Chem. Abstr., 1978, 88, 152668.
[13]CJ Cavalito, CM Martini and FC Nachod; Journal of American Chemical Society, 1951, 73, 2354.
[14]N Siddiqui and A Hussain; Indian Journal of Pharmacology, 2001, 33, 382-383.
[15]CW Pouton ; Euro.J.Pharma.Sci., 2006, 29, 278-287.
[16]DJ Faulkner ; Tetrahedron Lett., 1993, 38, 21-25.
[17]JJ Bhatt, BR Shah, PB Trivedi, NK Undavia and NC Desai, Ind. J. Chem, 1994, 33(B), 189-192.

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43
Synthesis of Substituted Flavanones and their Impact on Seed
Germinations
S. P. Rathod1*,R.T.Parihar2 ,A.P.Mitake3 ,T.M.Bhagat4 and
S. B. Waghmare5
1. Department of Chemistry, G. S. G. College, Umarkhed (MS) India.
2. Department of Chemistry,Vidnyan college,Malkapur (M.S) India.
3. Department of Chemistry, G. S. G. College, Umarkhed (MS) India.
4. Department of Chemistry, G. S. G. College, Umarkhed (MS) India
5. Department of Chemistry, G. S. G. College, Umarkhed (MS) India
E-mail – [email protected]

ABSTRACT:
As a part of systematic investigation, spectral analysis and morphology (root and shoot
elongation) of a several chlorosubstituted flavanones from diketones, gives various series by
using different aldehydes. As in series, 3-methoxy-8-chlorobenzoylflavanone,3-
chlorobenzoyl-8-chloroflavanone,3-benzoyl-8-chloroflavanone.
KEWORDS;- Diketones, flavanones, root and shoot elongation.
INTRODUCTION:
Flavanones are group of common natural polyphenolic compound that are widely found
in plant kingdom. It consists of two aromatic ring links through three Carbon bridge with a
carbonyl function. Actually these are the class of Flavanoids, which can be subdivided into
several classes such as chalcone, flavanones, isoflavanones, aurones etc. The flavanones are
mainly distributed in citrus fruitsIn present study, various chlorosubstituted flavanones were
synthesized from chlorosubstituted diketones and screened for their morphology (i.e. root and
shoot elongation) against some crop plants. Flavanones and their analogues having attracted
considerable attention because they possess antioxidant effect, cytotoxic, antimicrobial.
Antinflamatory activities etc.
Larget R et.al.1 reported substituted flavanone & studied as neuroprotective agents.
Venkatramn et.al2 synthesized the flavanone from chalcone by dehydrogenation with SeO2.
Yoigtandes et.al. 3 reported the flavanone from flavanone by heating with I2. Patill4 synthesized
flavanone from by using F2/DMSO as a dehydrogenating agent. Mayer A.M.et.al5 synthesized
cumarine and their role as growth regulators in several plants.. Korade D.L. et.al.6 reported the
effect of Anthracenes on seed germination of Lolium multiflorum plants. Gibba Z. et.al.7
studied the effect of nitric oxide on germination of Empress tree seeds.Majumdar, G. p.8
reported studied the cambial activities of root habit and shoot development in some plants.
The reaction of diketones with various aldehydes gives various flavanones these
reactions carried out in presence of ethanol as energy transfer medium in aq.KOH.
It has been well established that, the presence of chlorosubstituted ketones present in
flavanones, with the hope that the resulting molecules would exhibit promising root and shoot
elongation as shown in table and graph.
EXPERIMENTAL METHODS:
All the glassware’s used in the present work were of Pyrex quality. Melting points were
determined in open capillary and are uncorrected. Purity of compounds was monitored on silica
gel coated TLC plate. I.R. spectra were recorded on SHIMADZU Spectrophotometer in KBr

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palates. The analytical data of compounds were highly satisfactory. All the chemicals used
were of analytical grade. All the solvents used were purified by standard methods. Physical
characterization data of all the compounds are given in Table 1.
Result and discussion
The synthetic methods used in present work are given below along with their, IR data.
Acetylation of P- Chlorophenol:
P – Chlorophenol 50 ml was mixed with acitic unhydride 60 ml & unhydrus sodium
acetate 5 gm the mixture was refluxed for bout on 1 hrs. it was cooled and poured into cold
water. Acetated layer was separated & washed with water several times finally it was purified
by distillation and the distillate of compound was collected at about 2320C. Yield 75%, B.P.
2320C
2 – Hydroxy 5 – chloro-acetophenone:
When Phenyl acetate (50ml) was mixed with anhydrous AlCl3 (120gm) & heated at
1200C for 45 min on oil bath. The reaction mixture was decomposed with ice cold water
containing little hydrochloric acid to get crude ketone. It is purified by dissolving in acetic acid
& allowing the solution to fall drop by drop into cold water with stirring. A slightly yellow
powder was obtained having M.P. = 860C, Yield = 72%
2 – Benzoyloxy 5 - chloroacetophenone:
2 – Hydroxy 5- Chloroacetophenone (0.04 mol) & (8.59 ml) Benzoyl Chloride (0.05
mol) were dissolved in NaOH (10 %) (30 ml). The reaction mixture was shaken for about 45
min. The product thus separated filtered, washed with water followed by sodium bicarbonate
(10%) again with water. The solid product was crystallized from ethanol to obtain. 2 –
benzoyloxy 5-chloroacetophenone white crystal is obtained having M.P. - 870C, Yield - 80%.
1 – (2-hydroxyl Phenyl) 5 – Chloro- 3 – Phenyl – 1, 3 – Propanedione:
2 – Benzoyloxy 5 - chloroacetophenone (1.5 gm) was dissolved in (40 ml) dry
Pyridine. The solution was warmed up to 600C pulverized KOH (15gm) was added slowly with
constant stirring. After 4 hours of heating then reaction mixture was acidified by adding ice
cold HCl (1 : 1). The brownish yellow solid product thus separated was filtered, washed with
sodium bicarbonate solution (10%) and finally again with water. It was then crystallized from
ethanol-acetic acid mixture to get 1 – (2-hydroxyl phenyl) 5 – Chloro -3 – Phenyl – 1, 3 –
Propanedione. M.P:1210C, Yield – 75 %.
3 – benzoyl - 8 – chloro flavanone:
A mixture of 1 – (2-hydroxyl phenyl- 5 – Chloro 3 – Phenyl – 1, 3 – Propanedione
(0.01 mol) (3 gm) & Benzaldehyde (0.12 mol) ( 1 ml) was refluxed in ethanol (25 ml) and
KOH (1 gm) for 15 – 20 min descried to get compound M.P. - 2300C, Yield - 79%.
3 – Methory – 8-chloro benzoyl flavanone:
A mixture of 1 – (2-hydroxyl phenyl)-3 – phenyl – 1, 3 – propanedione (0.01 mol) (3
gm) & anisaldehyde (0.12 mol.) (1 ml) was refluxed in ethanol (25 ml) & KOH ( 1 gm) for 15
-20 min. & process as descried to get compound, 3 – methoxy-8 – chloro benzoyl flavanone.
M.P. - 2510C, Yield – 72 %.

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Table (1): Characterization data of synthesized new compound

Compound Molecular M.P. (oC) Yield (%) Rf
Formula
3a C21H13O3Cl 230 79 0.76
3b C22H16O4Cl 251 75 0.78
3c C21H14O3Cl2 240 75 0.79

TABLE NO. 1 : SEEDS - GRAM

Sr. Comp. % of germination Average length of root in cm. Average length of shoot in cm.
No.
Day Day Day

2 4 6 8 10 2 4 6 8 10 12 2 4 6 8 10 12

1 3a 80% 100% 1 2 2.4 3.2 3.5 4 0.6 0.9 2.8 3.2

2 3b 80% 100% 1 1.5 2 2.7 3.2 3.9 0.6 0.9 1.8 3.2

3 3c 100% 100% 2 2.4 2.6 3.4 3.7 4 0.6 0.9 1.4 2.9

4 Control 80% 90% 0.5 1.2 1.4 2 2.5 2.7 0.5 0.6 0.6 0.9

TABLE NO. 2 : SEEDS - Black Eyed Beans (Chawali)

Sr. Comp. % of germination Average length of root in cm. Average length of shoot in
No. cm.

Day Day Day

2 4 6 8 10 2 4 6 8 10 12 2 4 6 8 10 12

1 3a 80% 100% 2.1 2.6 2.9 3.2 4.5 5.4 0.6 0.9 2.8 3.2

2 3b 80% 100% 2.6 3.3 4.0 4.4 5.4 6.6 0.6 0.9 1.8 3.2

3 3c 80% 100% 2.6 3.3 4.0 4.4 5.4 6.6 0.6 0.9 1.8 3.2

4 Control 80% 90% 1.8 2 2.1 2.7 2.7 2.9 0.5 0.6 0.6 0.9

SEED : GRAM
8
7
ROOT ELONGATION

6
3a
5
3b
4
3 3c

2 Control
1
0
1 2 3 4 5 6
DAYS

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Conclusion
The present study was aimed at investigating the impact of newly synthesized
substituted flavanone on some crop plants viz: Gram and , Black Eyed Been (Chawali). The
choice of these crops was based on their enormously vast utility and also the indispensability
for the survival of the human race, all across the globe.
The efforts have been made to investigate and analyze the convergence and divergence
effect of test compounds on the morphology of plants under investigation. It was interesting to
note that, all the treated seeds exhibited remarkable roots and shoots elongation as compared
to untreated ones.
When the growth of all the treated plants were compared among themselves, it was
distinctly observed that, the change which is dominant while applying the treated compound
i.e. chlorosubstituted flavanones in Gram, Black Eyed Been (Chawali). In the initial stage
vegetative growth was not significance but after 2nd interval it gradually increases and after 12
days roots and shoots elongation were dominant to a considerable extent. Thus there has been
fair amount of satisfaction in crying out the present study. The encouraging results have surely
contributed to the enthusiasm of the author. But honestly, this is just the beginning. There is a
much scope for further study, and there is a long way to go.

ACKNOWLEDGEMENT
The authors are thankful to G.S.G.College, Umarkhed affiliated by S.G.B.Amaravati
University (M.S.) for providing all the facilities to carry out the work.
References:
1. Larget R, , B, Renard P, Bioorg med. Chem. Left 10(8) : 835-38, April 2000.
2. ., Venkataramn K, Mahal H.S, : Curr.Sci27, (1933), 124
3. Yoigttande, H.W., and Haertner : Arch. P(harm (YYeinneim Ger.) 316(3), (1983), 219-22.
4. Patil K.N., Ph.D. Thesis “Synthesis and rection of 3-aroylchoroflavanoneides.” Amravati
University (1993).
5. Mayer A.M., Poljakotta A, Amer Lowa Univ. Press 735-749, 1993.
6. Korade D.L. & Fulekan M.H., Biology and Medicine, Vol 1(1) : 28-34, 2009.
7. Gibba Z., Grubisic D., Todorovic S. Plant Growth Regulation 26, 175-181, 1998.
8. Majumdar, G.P. Hetero-archic roots in Enhydra fluctuans Laour. J. Indian bot. Soc. (11), 225-
227; 1932.
9. Rathod S.P.,Rajput P.R.,Rasayan J.Chem.4(3),660-665:2011.
10. Osman A.M.,Hasan K,Indian J.Chem.14B,282: 1954.
11. Rajora J,,Yadav J, Kumar R, Indian J.Chem.49B,989-993: 2010.
12. Kaliranjan R., Rathore S., Indian J.Chem.50B,1794-1799: 2011.

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44
Antimicrobial Resistance : A Global Challenge

Reshal Deshmukh
Department of Chemistry, Shri Shivaji Science College, Congress Nagar Nagpur
Email [email protected]

ABSTRACT
Decades after the first patients were treated with antibiotics, bacterial infections have again
become a threat because of the rapid emergence of resistant bacteria .Multidrug resistant
urinary tract infection is a most problematic infection and it is very difficult to doctors to
manage the patient by using appropriate antibiotics due to development of multidrug resistant
strains of bacteria . MDR strains shows resistant to maximum number of antibiotics and
sensitive to very few antibiotics.
Keywords : Urinary tract infection , multidrug resistant , antibiotic , bacteria
INTRODUCTION:
The bacterial associated Urinary Tract infection is most common infection found in all ages
and both the genders offering antibiotic resistance in most of the cases of infection. Now a
day’s antibiotic resistance has becomes major public health problem leads to increase resistant
to huge number of antibiotics from few years. The antibiotics are also responsible to develop
multidrug resistant strain due to resistance genes of resistant bacteria .The rapid emergence of
antibiotic resistant transfer genes among uro-pathogens are important cause of development of
antibiotic resistance. The increased antibiotic resistant suggest that the choice of antibiotic
should be guided by culture and sensitivity assay. In past decades many kind of resistant strains
are discovered.
PRINCIPLE OF ANTIMICROBIAL THERAPY
The objective of antibacterial therapy is to eliminate the bacterial growth in the urinary tract
and the antibiotics used are efficacious, safe, and cost effective. The basic c treatment starts
with the antibiotics which are number of choices available in market. The resolution of
infection is dependent on the susceptibility of the bacteria to the concentration of the antibiotic
achieved in urine. Antibiotics should eliminates the bacterial growth in urinary tract when
proper antibiotics are used which is able to achieve level in urine and the duration that this level
remains above the minimum inhibitory concentration (MIC) of microorganism that attack on
urinary tract. (Hooton TM, 1991). An effective antibacterial usually achieved MIC both in
serum and urine of healthy adults, the urinary level often many folds greater than serum level.
However the serum levels are critical in patients with uro-sepsis and urinary tract infection
involving renal parenchyma. (Jacobson SH, 1991)
Resistance to B-Lactams
Urinary bacteria may develop several mechanisms of resistance to Blactams which is most
efficient method of resistance to these agents in Gramnegatives is the synthesis of B-
lactamases. B- lactamases have presumably evolved to fight natural B-lactams, produced by
bacteria such as Streptomyces, Lysobacter , penicillium or Acremonium. (Brakhage AA,

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2005). However the widespread administration of antibiotics is influenced the development of

B- lactamases mediated resistance. Since B- lactam antibiotics came into clinical use, B-
lactamases are evolved with them. The emergence of MBL mediated resistant among
enterobacteriacae has also becomes serious health concern. More ever the increase in
international travel is likely to be contributory factor for the ascendancy of mobile MBL genes
as much as the mobility among individual bacteria.
Resistance to Colistin
Colistin belongs to the antimicrobial class of polymyxin and acts by binding to the lipid a
moiety of the bacterial lipopolysaccharide and subsequently disintegrating the bacteria
membrane. Resistance to colistin associated with lipopolysacharide modification leading to a
high level of antibiotic non-susceptibility
Resistance to Tigecycline
Tigecycline is first glycylcycline to be approved by the FDA, may lose its activity against MDR
strains, particularly when low drug concentration are attained in the serum during treatment.
Resistance develops when the MIC of the targeted pathogen exceed the Cmax of the drug,
which is almost the rule for all targeted A. baumannii strains. According to molecular studies
efflux pumps seem to be the most important mechanism of resistance to tigecycline both in
enterobacteriacae and non-fermenting rods.
Resistance to Fluoroquinolone
Fluoroquinolone such as ciprofloxacin with bactericidal activity against gram negative
pathogens. Fluoroquinolone act by binding to the nuclear enzyme DNA gyrase and
topoisomerase IV. In spite of particular targets, resistance mechanisms both chromosomal and
plasmidic have been developed. Chromosomal resistance mechanism includes target mutation
and augmented expression of efflux pumps.
CONCLUSION:
Antibiotic therapy is a choice of treatment for the bacterial infections , but due to development
of multidrug resistant strains the antibiotics are not effective as a treatment to patient. New
antibiotic developed which may be the used as a antibiotic therapy in future in resistant cases.
The present review is helpful to know few multidrug resistant pathogens are sensitive to
particular antibiotics and the proper antibiotic has been administrated to patient for rapid cure
of infection.
REFERRENCES :
 Brakhage AA, Al-Abdallah Q, Tuncher A, Sprote P. Evolution of B- lactam biosynthesis genes and
recruitment of transacting factors . Phytochemistry,2005 , 66(11),1200-1210.
 Dozzo P, Moser HE. New aminiglycoside antibiotics.Expert Opin. Ther. Path. ,2008 , 20(10), 1321-1341.
 Drlica K, Malik M,. Fluroquinolones : action and resistance. 2003 , Curr. Top. Med. Chem. 3(3) ,249-282.
 Falagas ME,Rafailidis PI ,Mathaiou DK. Resistance to polymyxin genes : Mechanisms , frequency and
treatment option. Drug resist. Update , 2010 , 13(4-5),132-138.
 Hooton TM , Stamm WE. Management of acute uncomplicated urinary tract infection in adults. Med Clin
North Am. 1991; 75:339-357.
 Jacobson SH . Afive year prospective followup of women with non-obstructive pyelonephritic renal
scarring. Scand J Urol Nephrl. ,1991;25:51-5
 Rawat D, Nair D. Extended spectrum B-lactamases in gram-negative bacteria. J Glob.infect Dis. ,2010,2(3)
,263-274

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45
Wings across Reservoirs: Understanding and Safeguarding the Migration
Patterns of Insectivorous Birds in Anthropogenic Landscapes
Dr. Chandrashekhar R. Kasar*, Ku. Revati Kishor Lonkar**
*Associate professor and Head, Department of Zoology, S.P.M. Gilani College, Ghatanji, Dist. – Yavatmal
Email – [email protected]
** Research Student, Department of Zoology, S.P.M. Gilani College, Ghatanji, Dist. – Yavatmal
Email – [email protected]

ABSTRACT:
The migration patterns of insectivorous birds in reservoir dams play a vital role in ecosystem
dynamics, influencing insect populations and contributing to overall biodiversity. This research
investigates the intricacies of bird migration in the specific context of reservoir dams, focusing
on insectivorous species and their seasonal movements. The study aims to understand the
routes, patterns, and ecological implications of bird migration in these anthropogenic
environments. The research delves into the ecological role of insectivorous birds in reservoir
dams. Preliminary analyses suggest a substantial impact on insect populations, with potential
cascading effects on the broader ecosystem. Understanding these ecological dynamics is
crucial for effective reservoir dam management and conservation. The study's findings have
implications for reservoir dam management, emphasizing the need for conservation strategies
that consider the ecological importance of these birds in maintaining ecosystem balance.
Keywords: Bird migration, insectivorous birds, reservoir dams, ecosystem dynamics.
INTRODUCTION:
Bird migration, a phenomenon deeply ingrained in the evolutionary tapestry of avian species,
plays a pivotal role in ecological systems worldwide. The intricate journeys undertaken by
birds across vast distances are essential for maintaining biodiversity, influencing insect
populations, and contributing to ecosystem resilience. In this context, our research focuses
specifically on the migration patterns of insectivorous birds within the unique environment of
reservoir dams.
1.1 Background: Reservoir dams, man-made structures designed primarily for water storage,
have become integral components of the human landscape. These environments, often shaped
by anthropogenic influences, present a distinctive ecosystem where wildlife, particularly avian
species, adapts to novel challenges. Insectivorous birds, a diverse group with varied migratory
behaviors, find themselves navigating these artificial landscapes.
1.2 Importance of Studying Bird Migration in Reservoir Dams: Understanding bird
migration in reservoir dams is not merely an academic pursuit but a necessity for effective
environmental management. Reservoir dams, with their altered landscapes and proximity to
human activities, pose unique challenges to avian species. Insectivorous birds, reliant on
specific habitats and food sources, become crucial indicators of the ecological health of these
environments.
As reservoir dams serve multifaceted roles, from water supply to energy production, their
impact on local ecosystems extends beyond their primary function. Our study recognizes the
need to explore how these human-altered landscapes influence the migratory behaviors of
insectivorous birds, species that contribute significantly to pest control and ecosystem balance.

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1.3 Specific Focus on Insectivorous Birds: The choice to concentrate on insectivorous birds
stems from their ecological significance. These avian species, characterized by their diet
primarily consisting of insects, fulfill essential roles in regulating insect populations. As they
traverse vast distances during migration, their interactions with the environment become
intricate ecological narratives. By studying the migration patterns of insectivorous birds, we
aim to unravel the complexities of their journeys and comprehend their contribution to the
ecological equilibrium of reservoir dam ecosystems.
1.4 Research Question and Objectives: At the heart of this study lies the fundamental
question: How do insectivorous birds navigate and contribute to ecosystem dynamics in the
context of reservoir dams? To address this question, our research objectives are as follows:
 To identify the insectivorous bird species inhabiting reservoir dam environments.
 To analyze the migration routes and patterns of these bird species.
 To investigate the influence of environmental factors, such as climate and food
availability, on the migration behaviors of these birds.
 To assess the ecological consequences of alterations in migration patterns within
reservoir dams.
This research seeks not only to enhance our understanding of bird migration in anthropogenic
landscapes but also to provide valuable insights for conservation strategies tailored to the
unique challenges presented by reservoir dam environments.
LITERATURE REVIEW:
2.1 Bird Migration Studies: Bird migration, a phenomenon studied for centuries, has been a
subject of fascination and scientific inquiry. Classical migration studies often focused on well-
known flyways, such as the East Atlantic Flyway or the East Asian-Australasian Flyway,
tracking the movements of waterfowl and shorebirds. While these studies have significantly
contributed to our understanding of long-distance migration, the dynamics of migration within
human-altered landscapes, such as reservoir dams, remain relatively understudied.
2.2 Migration in Anthropogenic Environments: The impact of human activities on bird
migration patterns has gained attention in recent decades. Urbanization, deforestation, and
climate change have altered the traditional routes and behaviors of migratory birds. However,
the specific influence of reservoir dams, with their unique combination of altered habitats and
human activities, merits closer examination. Some studies have explored how artificial water
bodies affect bird migration, but the focus on insectivorous species within reservoir dams
remains limited.
2.3 Significance of Insectivorous Birds in Ecosystems: Insectivorous birds, a diverse group
encompassing species from various families, play crucial roles in maintaining ecological
balance. Their diet primarily consists of insects, making them natural regulators of pest
populations. In agricultural landscapes, insectivorous birds contribute to integrated pest
management, reducing the need for chemical interventions. However, the extent of their
contribution in reservoir dam ecosystems and the potential consequences of altered migration
patterns remain underexplored.
2.4 Environmental Factors Influencing Bird Migration: Numerous environmental factors
influence bird migration, including climate conditions, food availability, and habitat changes.
Climate change, in particular, has been identified as a significant driver of alterations in
migration patterns. Understanding how these factors intersect within the context of reservoir

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dams is crucial for predicting and mitigating potential ecological disruptions. The literature
review seeks to identify key studies that have examined the interplay between environmental
factors and bird migration in both natural and artificial environments.
2.5 Gaps in Current Knowledge: Despite the wealth of information available on bird
migration, there is a noticeable gap in literature concerning insectivorous birds in reservoir
dams. Existing studies often focus on waterfowl or larger species, neglecting the ecological
nuances associated with insectivorous behaviors. This literature review underscores the need
for targeted research that specifically addresses the migration patterns, behaviors, and
ecological roles of insectivorous bird species within reservoir dam environments.
METHODOLOGY:
3.1 Study Sites Selection: The choice of study sites is a critical aspect of our methodology.
Reservoir dams selected for this research are characterized by diverse ecological settings,
representing different climatic zones and geographic regions. This diversity ensures a broad
understanding of how insectivorous birds adapt to various environmental conditions within
reservoir dams.
3.2 Field Observations and Behavioral Analysis: Continuous field observations will
complement the technological aspects of our methodology. Researchers will spend extended
periods at the selected study sites, documenting bird behaviors, feeding patterns, and
interactions with the reservoir dam environment. Behavioral analysis will focus on
understanding how insectivorous birds adapt to the altered habitats presented by reservoir
dams, including changes in foraging behavior and nesting preferences.
3.3 Climate and Habitat Analysis: Environmental factors, such as climate conditions and
habitat changes, are integral components of our study. Climatic data will be collected to
correlate with bird migration patterns, exploring the influence of temperature, precipitation,
and seasonal variations on migration behaviors. Habitat analysis involves assessing the
availability of insect prey, vegetation cover, and water quality within reservoir dams.
3.4 Data Analysis: The collected data will undergo rigorous analysis. Statistical methods will
be applied to identify migration hotspots, determine seasonal variations in bird populations,
and assess correlations between environmental factors and migration behaviours. Geospatial
tools will aid in mapping migration routes and stopover locations.
3.5 Ethical Considerations: All aspects of the research adhere to ethical standards and
prioritize the well-being of the studied birds. Bird handling follows established protocols to
minimize stress and potential harm. The research team has obtained necessary permits and
approvals from relevant wildlife conservation authorities, ensuring compliance with legal and
ethical guidelines.
3.6 Community Engagement: Recognizing the importance of community involvement, our
methodology includes outreach programs to engage local communities residing near the
selected reservoir dams. Workshops, awareness campaigns, and citizen science initiatives aim
to foster a sense of shared responsibility for the conservation of insectivorous birds and their
habitats.
3.7 Limitations: While our methodology is comprehensive, certain limitations are inherent in
avian migration studies. External factors such as predation, disease, and unforeseen events may
impact the reliability of data collected through bird banding and satellite tracking. However,
the combination of multiple data sources enhances the robustness of our findings.

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CASE STUDIES:
4.1 Case Study 1: The Warbler's Journey Species: Black-and-white Warbler (Mniotilta
varia) Migration Route: Northeastern study site to Central America Key Findings: This case
study reveals a remarkable migratory journey of Black-and-white Warblers navigating from a
northeastern reservoir dam to their wintering grounds in Central America. Satellite tracking
data indicated distinct stopover points in areas with abundant insect populations, emphasizing
the critical role of reservoir dam habitats in supporting migratory journeys.
4.2 Case Study 2: Navigating Urban Reservoirs Species: Common House Martin (Delichon
urbicum) Migration Route: Urban reservoir dams in European settings Key Findings: In
contrast to traditional rural habitats, this case study explores the migratory patterns of Common
House Martins within urban reservoir dams. These insectivorous birds demonstrated adaptive
behaviors, utilizing artificial structures for nesting and displaying adjusted migration routes
influenced by urban landscape features.
CONSERVATION STRATEGIES:
5.1 Habitat Restoration and Enhancement: Implementing habitat restoration initiatives
within reservoir dams involves reestablishing native vegetation, maintaining suitable nesting
sites, and creating insect-rich environments. This strategy aims to ensure that migratory birds
find optimal conditions for feeding, resting, and breeding.
5.2 Climate-Responsive Conservation: Given the influence of climate on bird migration,
conservation efforts must be adaptive to changing climatic conditions. Integrating climate-
responsive strategies involves monitoring climate trends, understanding their impact on bird
behavior, and adjusting management practices accordingly.
5.3 Community-Based Conservation: Engaging local communities in the conservation
process is paramount. By fostering awareness, providing education, and involving
communities in citizen science initiatives, a sense of shared responsibility is cultivated. Local
support contributes to sustainable conservation practices and the protection of reservoir dam
ecosystems.
5.4 Sustainable Development Practices: Promoting sustainable development around
reservoir dams ensures the coexistence of human activities and wildlife conservation.
Implementing regulations that minimize disturbances, control pollution, and prioritize
ecosystem health contributes to the long-term viability of insectivorous bird populations.
CONCLUSION:
This research has delved into the intricate world of insectivorous bird migration within
reservoir dams, uncovering diverse patterns, adaptive behaviors, and ecological interactions.
The case studies underscore the importance of these avian species in maintaining ecological
balance and pest control within human-altered landscapes.
The conservation strategies proposed emphasize a holistic approach that integrates habitat
restoration, climate-responsive measures, community engagement, and sustainable
development practices. Preserving the migratory routes of insectivorous birds is not only vital
for their well-being but also contributes to the overall health and resilience of reservoir dam
ecosystems.
As we navigate an era marked by rapid environmental changes, understanding and
safeguarding the migratory journeys of insectivorous birds become imperative. This research
serves as a stepping stone towards comprehensive avian conservation strategies, bridging the

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gap between scientific knowledge and on-the-ground initiatives. The journey of these birds,
intertwined with the ecosystems of reservoir dams, unveils a narrative of adaptation, resilience,
and the delicate balance between human activities and the natural world.

WORKS CITED:
1. Ahmed, N., Khan, M. S., & Rahman, K. U. (2018). "Biodiversity Assessment of Birds
in Urban Reservoirs of Karachi, Pakistan." Pakistan Journal of Zoology, 50(3), 1019-
1027.
2. Alerstam, T., Hedenström, A., & Åkesson, S. (2003). "Long-distance migration:
evolution and determinants." Oikos, 103(2), 247-260.
3. BirdLife International. (2021). "Delichon urbicum (amended version of 2020
assessment)." The IUCN Red List of Threatened Species.
4. Gill, R. E., Douglas, D. C., & Handel, C. M. (2001). "Long-term tracking of arctic terns:
new global information on migration pathways and breeding grounds." The Condor,
103(2), 428-440.
5. Newton, I. (2008). "The Migration Ecology of Birds." Academic Press.
6. Robinson, R. A., Learmonth, J. A., & Hutson, A. M. (2003). "Atlas of Bird Migration:
Tracing the Great Journeys of the World's Birds." Random House UK.
7. Runge, C. A., Martin, T. G., Possingham, H. P., Willis, S. G., & Fuller, R. A. (2014).
"Conserving mobile species." Frontiers in Ecology and the Environment, 12(7), 395-
402.
8. Saini, J., & Khera, N. (2017). "Assessment of Water Quality in Reservoirs of Shimla
Hills Using CCME Water Quality Index." Journal of Scientific Research & Reports,
14(3), 1-14.
9. Sanderson, F. J., Donald, P. F., Pain, D. J., Burfield, I. J., & van Bommel, F. P. (2006).
"Long-term population declines in Afro-Palearctic migrant birds." Biological
Conservation, 131(1), 93-105.
10. Wang, Y., Zhu, W., Zhang, Y., & Ma, M. (2020). "Impacts of Urban Reservoirs on the
Distribution and Diversity of Wintering Waterbirds." Water, 12(1), 176.
11. Zalles, J. I., & Bildstein, K. L. (2000). "Raptor Watch: A Global Directory of Raptor
Migration Sites." BirdLife International.

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46
Synthesis and Characterization of Pyrazolone and its derivatives.
Mr. R. N. Gaikwad
Maharashtra Udayagiri Mahavidyalaya,Udgir Dist.Udgir 413517
Abstract:
The synthesis of pyrazolone and its derivatives is achieved by a clean, one-pot, multi-
component condensation reaction combining ethyl acetoacetate, hydrazine hydrate, aromatic
aldehyde in catalytic amount of MSA. This approach has numerous benefits, including a quick
reaction time and an easy work-up process. Compounds were identified via mass spectral
analysis, NMR, IR, and chemical transformation.
Keywords: aryl aldehyde, ethyl acetoacetate, hydrazine hydrate, MSA, pyrazolone etc.
Introduction
Pyrazolone are a class of organic compounds that feature a five-membered ring containing
three carbon atoms and two nitrogen atoms. The molecular formula of pyrazolone is C 3H3N2.
This heterocyclic ring structure imparts unique chemical and pharmacological properties to
pyrazolone, making them significant in various fields such as chemistry and medicine.
Pyrazolone also exhibit interesting coordination chemistry, forming complexes with various
metal ions. This property makes them relevant in coordination chemistry and catalysis.The
chemical, pharmaceutical, and agrochemical industries are very interested in pyrazolone
derivatives1,2. Considered an active scaffold with pharmacological importance, pyrazolone and
its derivatives exhibit nearly all pharmacological activity. The pyrazolone moiety's
pharmacological potential has been demonstrated by the nucleus's presence in pharmacological
agents belonging to various therapeutic categories, including rimonabant, difenamizole, an
analgesic; fezolamide, an antidepressant; and celecoxib, a potent anti-inflammatory and
antipsychotic.3 Because of their many characteristics and uses, pyrazolones are a class of
chemicals that have been studied. These include pyrazol-3-ol and pyrazolin-5-one.
Sulfamazone, propyphenazone, and nifenazone4 are a few of the exploratory small compounds
containing pyrazolone that have been considered as potential therapeutic possibilities.
Derivatives of pyrazolone are a significant class of heterocyclic chemicals found in numerous
medications and synthetic goods. Based on a review of the literature, pyrazolone derivatives
exhibit a range of pharmacological actions, including, anti-tubercular5, anti-fungal,6,7 anti-
bacterial,8 anti-inflammatory,9 anti-timor,10 anti-microbial.11
In this study, we have created a sophisticated, effective, simple, and straightforward method
for the synthesis of pyrazolone and its derivatives. We do this by employing ethanol as a solvent
together with hydrazine hydrate, ethyl acetoacetate, and substituted aryl aldehydes in a catalytic
quantity of MSA. In terms of environmentally friendly procedures, the combination of the
green technique and the catalyst results in an outstanding product yield. This approach offers
many important benefits, including reduced reaction time, solvent-free conditions, atom
economy, using a green catalyst, and a more pleasant workup process.
Experimental
General
The digital melting point device (Veego-VMD) was used to record the uncorrected melting
point of the produced chemical. A.R. Grade chemicals were used. Every chemical employed
was of A.R. Grade, and it wasn't purified further. Microwave irradiation was carried out in a
Microwave Oven, Model No. MS2079 DB DB1QILN (2450 MHz, 1050 W) equipped with

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Erlenmeyer flask. The compound's melting point was measured in an open capillary tube and
might not be accurate. Using an FT-IR spectrophotometer made by Perkin-Elimer, IR spectra
were recorded. KBR Pellet. Using chloroform as a solvent, the 1H NMR spectra and mass were
recorded using a Perkin-Elimer spectrophotometer. The chemical shift value was reported in
parts per million (ppm) with respect to TMS, which served as an internal standard.
General Procedure for the synthesis of Pyrazolone and its Derivatives
The combination of 1ethyl acetoacetate (0.01mmol), 2hydrazine hydrate (0.01mmol), and 3
aryl aldehyde (0.01mmol) in catalytic amount of Methane sulphonic acid (MSA) in an
Erlenmeyer flask and irradiated until completion of the reaction in microwave oven.The
reaction mixture was poured in crushed ice and filtered, washed with ethyl acetate, the product
was recognized as 3-methyl-4-arylidene pyrazolone (4a-d). The crude product was collected
and recrystallized in ethanol and dried. The entire product was characterized by physical
constant and spectroscopic techniques and compared with the standard method.

Scheme 1
Result and Discussion
As indicated in the scheme 1, we reported a direct method for the synthesis of pyrazolone and
its derivatives by one pot condensation of aromatic aldehyde, hydrazine hydrate, and ethyl-
acetoacetate.The yield was really high. In order to assess the extent and constraints of the
methodology, the reactions were conducted in the absence of a catalyst, and the structures of
all newly synthesized pyrazolone derivatives were established using chemical transformation,
NMR, IR, and mass analysis.
Table1:Synthesis of pyrazolone & its derivatives catalyzed by MSA under M.W.
(120oC/300W)

Product Name of compound % yield Melting Point in OC

4a (Z)-3-benzylidene-4-methyl-1,3- 94% 122 0C
dihydro-2H-pyrrol-2-one
4b (Z)-3-(4-chlorobenzylidene)-4-methyl- 89% 215 0C
1,3-dihydro-2H-pyrrol-2-one
4c (Z)-4-methyl-3-(3-nitrobenzylidene)-1,3- 87% 195 0C
dihydro-2H-pyrrol-2-one
4d (Z)-3-(4-hydroxybenzylidene)-4- 92 197 0C
methyl-1,3-dihydro-2H-pyrrol-2- %
one

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Table 2. Optimization of catalyst concentration of (4b) under M.W. Irradiation

Entry Catalyst mol % Time, min Yields %
1 No catalyst 9 -
2 1 8 76
3 2 7 89
4 3 3 95
5 4 5 85
6 5 8 70
Initially, the mixture of Ethyl acetoacetate, Hydrazine hydrate and substituted aryl aldehydes,
(0.01 mol each) was taken in an 50 mL vessel and subjected to microwave irradiation under
neat conditions in the absence of catalyst and solvent, and the reaction was failed to get the
product under this condition. The reaction was repeated in the presence of catalyst in the range
1-5 mol %. We have optimized the amount of catalyst for the synthesis of (4b, Table 1) under
microwave irradiation and excellent yield was obtained in only 3 minutes when 3 mol %
catalysts are introduced (Table 2, entry 4).
Spectroscopic characterization.
4c) (Z)-4-methyl-3-(3-nitrobenzylidene)-1,3-dihydro-2H-pyrrol-2-one
Solid: - Melting point = 195oC
IR (KBr, Cm-1) : 1354, 1260, 1500, 1607, 670.
1
HNMR (400 MHz, DMSO-d6):δ 7.2(1H,s), 1.18 (1 H, s), 6.93 (1 H, s), 8.10-8.12 (4H, m).
4d) (Z)-3-(4-hydroxybenzylidene)-4-methyl-1,3-dihydro-2H-pyrrol-2-one
Solid: - Melting point = 197oC
IR (KBr, Cm-1) : 1354, 1260, 1500, 1607, 670.
1
HNMR (400 MHz, DMSO-d6): δ 7.45(1H,s), 6.89 (1 H, s), 9.81 (1 H, s), 5.0(1H,s), 6.93 (4H, m).

Conclusion
We have been able to offer an effective method for the synthesis of pyrazolone derivatives in
the presence of MSA in our current study. All of the recently synthesized compounds' structures
were determined via mass spectral analysis, NMR, IR, and chemical transformations.
Condensation-enhanced pyrazolone synthesis and its derivatives provide high yields of
pyrazolone in good quality and purity at a low cost.. This approach has the noteworthy
advantages of being catalyst-free and having an easy workup procedure. The synthesis process
used to create these molecules is very easy to use, clean, economical, green, and
environmentally friendly.
References :
1. S. Mert, R. Kashmogullari, Postdoc, 2, 450, 2014.
2. G.Mariappan, B.Saha, Journal of pharmacy research, 3, 2856, 2010.
3. P. Gupta, K.Halve, International journal of pharmaceutical sciences and research, 4, 2291, 2015.
4. Z. Zhao, X. Dai, Eurj.Med.Chem, 186, 1118, 93, 2020.
5. K. Karrouchi, S. Radii, Y. Ramli, Molecule, 23, 45, 2018.
6. M.Radii, V. Bernardo, B. Bechi, D. Castagnoio, Tetrahedron letters, 50, 6572, 2009.
7. P. Mitra, A.Mitra, Indian Chem. Soc., 58, 695, 1981.
8. B.Singh, Indian Chem. Soc., 68, 165, 1991.
9. M. Ruoqun, J. Zhu, J. Liu, L.Chem, X. Shen, H. Jiang, Molecule, 15, 3593, 2010.
10. R. Prajuli, J.Banerjee, H. Khanal, Oriental journal of chem., 31, 540, 2015.
11. M. Nashwa El- Metwaly, G. Marwa, A. Thoraya, M. Abelaua, Biogenetic chemistry and applications, 3,
654, 2018.
12. British Pharmacopici Vol. 2, Her Mejestis Stationary Office, London,12,(1980)British Pharmacopeia’s
Pharmaceutical press London,(1953)796.
13.Bary A.I., The antimicrobial susceptibility test, Principle and Practices Illusiea and Febiger, Philadelphia,
PA, USA, 180.
14.Cavangh F., Analytical Microbiology Academic Press New York, (1963) 126.

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47

Preliminary Phytochemical analysis of Leaves extract of the plant Justicia

Adhatoda
Swapnil D. Bhagat1*, Sopan D. Ingole1, Varun A. Mahale2, Nandkishor S. Thakare1&
Chandrakant U. Dhanwad1
1
Department of Chemistry, M.S.P. Arts, Science & K.P.T. Commerce College, Manora, Maharashtra, INDIA
2
Department of Chemistry,S.S.S.K.R. Innani Mahavidyalaya Karanja (Lad), INDIA
[email protected]
Mobile No. 9595197970
Abstract:
The well-known Indian medicinal herb Justicia adhatoda is knowned for its pharmacopoeia.
This plant has been used commonly in the ayurvedic system of medicine. This plant's root,
bark, leave and flower are used to heal many types of infection. In the present study, we have
done the phytochemical screening of its leaves for glycoside, flavonoids, tannin, alkaloid,
phenols, terpenoids and steroids and in the prepared hydroalcoholic extracts using Soxhlet
extraction method
Key Words: Phytochemical Analysis, Justica Adhatoda plant.

Introduction:-
Justicia adhatoda is a member of family Acanthaceae which is a small evergreen herbal plant.
This plant is distributed all over the India. All the parts of this plant has been used for their
curative effects from ancient times1. J. adhatoda is a widely used plant medication in Unani
and Ayurvedic medicin2. Justicia adhatoda produces vasicine, which is used to treat a variety
of illnesses, primarily respiratory tract conditions. In Sweden J. adhatoda is classified as a
natural remedy and some preparations based on protocols against cough containing an extract
of J. adhatoda are accessible3.
It is used by Ayurvedic physicians and possesses some medicinal properties. It has been used
to treat a wide range of illnesses, most notably those affecting the respiratory system. As a
result, it is one of the main herbs in the Ayurvedic system and is used to treat asthma, bronchitis,
cough, and cold symptoms4. It has been used to control both internal and external bleeding
such as peptic ulcers, hemorrhoids, bleeding gums and also used for a multitude of disorders
including; leprosy, blood disorders, heart troubles, fever, vomiting, loss of memory,
leucoderma, jaundice, tumors, mouth troubles, sore-eye and gonorrhea. In Sri Lanka, the plant
is used to cure menorrhagia and profuse phlegm5. It is also used for the treatment of bleeding
piles6, impotence and sexual disorders7.
In order to prepare the respiratory tracts for the rigorous breathing exercises, chewing the leaf
buds alone or with a small amount of ginger root is a common yogic practice. Many leaf
preparations are used in Southeast Asia to treat leprosy, bleeding, haemorrhage, skin
conditions, wounds, and headaches8,9. The bruised fresh leaves are used for snake-bites in India
and Sri Lanka10.

Methods and Materials:

The plant leaf extract was prepared by using 25 gm of fresh leaves collected from the local
areas. Fresh leaves were washed extensively with water followed by final wash twice thrice
with distilled water to remove all the dust and unwanted visible particle. The leaves were cut
in to small pieces and then shade dried for 2-3 days. Leaves were homogenized to a fine coarse
powder using mortar and pestle and then stored in fine air tight container for further process.

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Preparation of Plant Extract:

The powdered dried plant leaves were taken in a round bottom flask and extracted (25g)
exclusively with 100 ml each of acetone, methanol, ethyl acetate, chloroform and diethyl ether
in a soxhlet extractor for 4 hrs. and few porcelain pieces were added in to it to avoid bumping.
After heating gently removed the solvent from sample by evaporation extract remains in round
bottom flask used for phytochemical test.

Phytochemical Analysis:
Phytochemical analysis was carried out using standard methods11,12,13,14.
1) Test for Carbohydrates :
Test for Starch:
Procedure: Plant extract + 5 ml 5% KOH
Observation: A canary colouration.
2) Test for Glycosides :
i. 10% NaOH test:
Procedure: 1 ml dil. H2SO4+0.2 ml extract+ boiled for 15 min + allowed
cooling+ neutralize with 10%NaOH + 0.2 ml feeling solution A and B.
Observation: A brick red precipitate.
ii. Concentrate H2So4 test:
Procedure: 5ml extracts solution + 2 ml Glacial acetic acid + a drop of 5%
Fecl3 + Con. H2SO4
Observation: A brown ring is observed
3) Test for Proteins and Amino acids :
Xanthoproteic test:
Procedure: Plant extract + few drops of Con. nitric acid.
Observation: Yellow colored solution.
4) Salkowski’s Test (Detection of Phytosteols):
Procedure: plant extract with chloroform + few drops of Conc. H2SO4, (Shake well
and allowed to Stand).
Observation: Formation of Brown Color ring.
5) Salkowski’s Test (Detection of Terpinoids):
Procedure :Plant extract + Chloroform + few drops of Conc. H2SO4, Shake well and
allowed to Stand.
Observation: Reddish brown coloration (at bottom).
6) Test for Alkaloids:
i. Wagner’s Test:
Procedure: 1 ml Plant extract + 2ml Wagner’s reagent (Solution of Iodine in
Potassium Iodide)
Observation: Brown colored ppt. is formed
ii. Mayer’s test:
Procedure: 1 ml plant extract with dil HCL +1-2 drops of mayer’s reagent
(Potassium mercuri iodide solution) Along the side of test tube.
Observation: yellow precipitate formed
7) Test for Flavonoids:
Lead acetate Test:
Procedure: Plant extract is dissolved in 5 ml of distilled water + 3 ml of 10% lead
acetate solution
Observation: A White ppt is formed

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8) Test for Phenol:

Iodine test:- Procedure: 1 ml extract+ few drops of dil. Iodine solution.
Observation: A red colour present
9) Test for Saponin:
Foam test Procedure : 0.5 gm plant extract+ 2 ml water (vigorously shaken)
Observation: Persistent foam for 10 min
Result and Discussion:
The results for the phytochemical analysis of Justicia adhatoda plant was shown in Table
no.1
Sr. No. Test Inference
1. Test for Caebohydrates (Test For Starch) +ve
2. Test for Proteins Xanthoproteic test +ve
3. Test for Phytosteols Salkowski’s Test +ve
4. Test for Terpinoids Salkowski’s Test +ve
Wagner’s Test +ve
5. Test for Alkaloids
Mayer’s test +ve
NaOH test +ve
6. Test for Glycosides
Conc. H2SO4 test +ve
7. Test for Flavonoids Lead acetate test +ve
8. Test for Phenol Iodine Test +ve
9. Test for Saponin Foam Test +ve

The result from table no.1 shows that among the analyzed phytochemicals are
Carbohydrates, Proteins, Phytosterols, Terpenoids, Alkaloids, Flavonoids, Saponin and
Phenols were found present in the ethanolic extract of leaves of Justicia adhatoda plant. The
medicinal properties of the plants are due to the presence of different phytochemicals which
are present in the leaves of the plant.
Conclusion:
From the result, it is concluded that extract of leaves of Justicia adhatoda plants contains many
active phytochemicals. Therefore the extract of these plants can be used as a drug. Further
studies on isolation, purification, and characterization of phytochemicals are suggested for the
development of new plant base pharmaceuticals having lesser side effects.

References:

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review of ethnopharmacological and toxicological data. J. Ethnopharmacol., 72: 1-20
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(1998) pp. 109-132,

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4. Karthikeyan A, Shanthi V, Nagasathya A (2009). Preliminary Phytochemical and

antibacterial screening of crude extract of the leaf of Adhatoda vasica (L). Int. J. Green
Pharm., 3: 78-80.
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pharmacological overview on Adhatoda zeylanica. Medic. Syn. A. vasica (Linn.) Nees.
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7. Pushpangadan P, Nyman U, George V (1995). Glimpses of Indian Ethnopharmacology.
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8. Adnan M, Hussain J, Shah MT, Ullah F, Shinwari JK, Bahadar A, Khan AL (2010).
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regions in Northwest Pakistan. J. Med. Plant Res., 4: 339-345
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Medica. Hamdard Foundation Press, Karachi, 1: 181-187.
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11. Handayani R. Harahap U., Karsono (2017): Hypoglycemic Activity of Nano Particles
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12. Doughari JH (2012): Phytochemicals:extraction methods, basic structures andmode of
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(3rdedn) eds. Chapman and Hall.
14. Trease GE, Evans WC (1989):Pharmacology, (11thed) Bailliere TindallLtd, London.

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48

In-Silico Prediction of Phytoconstituents From

Solanum Indicum for Antistress Activity Targeting Ask 1 Inhibitor
Sakshi Mude*, parimala Katolkar, Pradeep Raghatate, Jagdish Baheti
Kamla Nehru College of Pharmacy, Butibori, Nagpur 441108 (MS), INDIA
*Email:[email protected]

ABSTRACT
Objective Stress in psychology is defined as tension or pressure on emotion. Natural
products and their active principles as sources for new drug discovery and treatment of diseases
have attracted considerable attention of researchers. Compounds found in medicinal plants
have been the source of many conventional medications. In-silico testing of Solanum indicum
phytoconstituents on ASK 1 inhibitor for antistress efficacy was a part of our investigation.
Methods
Utilizing Discovery studio, molecular docking is done to assess the pattern of interaction
between the phytoconstituents from the plant Solanum indicum and the crystal structure of the
ASK 1 inhibitor (PDB ID: 3VW6). Later, SwissADME and pkCSM were used to screen for
toxicity as well as the pharmacokinetic profile. Further, interaction between proteins by
STRING network analysis was also evaluated.
Results
The docked results suggest that Solafuranone (-8.4 kcal/mol), Isofraxidin (-6.9 kcal/mol) for
3VW6 macromolecule has best binding towards antistress activity as compared to the standard
(Fluvoxamine) for 3VW6 is -6.6 kcal/mol. Furthermore, pharmacokinetics and toxicity
parameters were within acceptable limits according to ADMET studies.
Conclusion
Targeting ASK 1 inhibitor against the stress by phytomolecules from Solanum indicum can
serve as a rational approach for designing future antistress drugs.
KETWORDS: In-silico, Solanum indicum, anti-stress activity, 3VW6, Discovery studio.

1. INTRODUCTION
The plant Solanum indicum, is a member of the Solanaceae family and is also known as Byakur,
Guta started, Kata began and Indian Night Shade. Solanum indicum, also known as Badi
Bhatkataiya in Hindi and "Brihati" in Sanskrit, is a plant used either alone or in combination
with other drugs in the Ayurvedic medicinal system. The use of seeds, roots, leaves, and berries
can be used to treat a variety of illnesses, including bronchitis, asthma, dry cough, rhinitis,
dysuria, leucoderma, sexual dysfunction, insomnia, weak heartbeat, and pruritis.1
A prickly perennial under shrub that can reach a height of 1 m and is mostly found in warmer
areas of India up to a height of 1500 m, Solanum indicum, sometimes known as poison berries
in English, is recognised for its poisonous berries-like fruit. This important medicinal plant is
frequently used to treat poisonous affections, skin diseases, ulcers, breathing difficulties,
abdominal pain, cough, and dyspepsia in folk and traditional Indian systems of medicine.
Siddha and Ayurveda both use it.2
However, there are few studies on the phytoconstituents of Solanum indicum for the antistress
activity. Thus, keeping the above information in view, the present investigation was designed
to identify the potential phytochemicals of Solanum indicum against 3VW6 using a molecular
docking method.

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2. MATERIALS AND METHODS

2.1. Platform for molecular docking
The computational docking study of all the phytoconstituents selected as ligands with antistress
activity as the target was performed using PyRx software.3
2.2. Protein preparation
The macromolecule is 3VW6, in silico analysis of selected phytoconstituents was performed
on the 2.40 Å crystal structure of antistress macromolecule with inhibitor, (PDB ID: 3VW6,
having resolution Resolution: 2.40 Å, R-Value Free: 0.252, R-Value Work: 0.213, R-Value
Observed: 0.215), which was retrieved from the protein data bank (https://www.rcsb.org).
3VW6 is classified as Crystal structure of human apoptosis signal-regulating kinase 1 (ASK1)
with imidazopyridine inhibitor all other molecules, such as co-crystallized water molecules,
unwanted chains, and nonstandard residues, were deleted. Using Discovery studio.4
2.3. Mechanism of Action
3VW6: Apoptosis signal-regulating kinase 1 (ASK1) was originally identified as a member
of the mitogen-activated protein kinase (MAP3K) family that activates both p38 MAP kinase
and c-Jun N-terminal kinase (JNK) pathways ASK1 is stimulated by various cell stressors
including cytotoxic cytokines, reactive oxygen species (ROS), and endoplasmic reticulum
stress. Recent studies revealed that ASK1 contributes not only to the regulation of cell death,
but also to cytokine responses, cell differentiation, and immune regulation. Therefore, ASK1
inhibitors are thought to have potential for the protection of cells from various stresses in
wide-ranging pathological situations such as autoimmune disease, diabetes, cardiovascular
disease, neurodegenerative disorders, and inflammatory disorders.5
2.4. Ligand preparation
The three-dimensional (3D) structures of all constituents were retrieved using Avogadro
software from the PubChem database available on the NCBI website
(https://pubchem.ncbi.nlm.nih.gov/). However, the drawing of geometrical 2D structure was
performed using the ChemSketch program. The two-dimensional (2D) structures were
transformed into 3D models using the Avogadro software and the ligand structures were saved
in the PDB format. All the chemical structures are shown in Figure 1.

Solanidine Solasodine
OH H
H3C
H3C NH CH3
H3C H
H3C H
CH3
HH
H O
CH3
N H

CH3 HO

Solavetivone Solafuranone
CH2 H3C
CH3 H3C O
O
CH3
O H3C CH3

CH3

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Scopoletin N-p-trans-Coumaroyltyramine

HO
H
N

O
OH

Isofraxidin Lauric acid

H3C O HO CH3

O
HO O O
O
H3C

𝛽-sitosterol Diosgenin
H
O
H3C

H3C H
H
H
O H

O H
H

Fig. 1. Chemical structures of all selected phytoconstituents in the molecular docking

studies
2.5. Standard Preparation
The standard is prepared steps such as, the 2D structure of standard drug was made using chem
sketch program, then the 2D structure was converted into 3D model using Avogadro Software,
it was saved in PDB format.
By using PyRx molecular docking of Fluvoxamine was done with 3VW6.
2.6. Molecular docking
Molecular docking evaluates the protein-ligand interactions and estimates the scoring function
based on the geometry to predict the binding affinity of the ligand molecule6,7. We
applied molecular docking studies to investigate the binding pattern of selected
phytoconstituents (Figure 1) and the standard drug, along with the crystal structure of antistress
activity macromolecule (PDB ID: 3VW6). The molecular docking study was performed using
PyRx software, Binding affinity was explored using the Vina wizard tool. The final results
were analysed and visualized using Discovery Studio 2020 Client 8, with bound ligands as the

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standard. Visualization of protein ligand interaction reflects the number of interactions and
active residues responsible for significant binding at the active site of the target enzyme.
2.7. Absorption, distribution, metabolism, and excretion (ADME) and toxicity prediction
The selected phytoconstituents and standard drug were further checked for drug-likeness
properties according to Lipinski’s rule. During drug development, it is necessary to predict the
tolerability of phytochemicals before being ingested by humans and animal models.
The pharmacokinetic profile (ADME) and toxicity predictions of ligands were conducted using
SwissADME (http://www.swissadme.ch) and pkCSM (an online server database predicting
small-molecule pharmacokinetic properties using graph-based
signatures, (http://biosig.unimelb.edu.au/pkcsm/prediction). To analyse the toxicological
properties of ligands, Simplified Molecular Input Line Entry System (SMILES) notations or
PDB files were uploaded, followed by selecting the required models for generating numerous
information about structure-related effects9,10.
3. RESULT AND DISCUSSION
The present study aimed to explore the inhibitory potential of the phytoconstituents present
in Solanum indicum targeting antistress activity. In this study, we performed molecular
docking studies of all phytoconstituents found in Solanum indicum using AutoDock Vina,
followed by a study of interacting amino acid residues and their influence on the inhibitory
potentials of the active constituents. Selected phytoconstituents showing the best fit were
further evaluated for absorption, distribution, metabolism, excretion, and toxicological
(ADMET) properties using SwissADME and pkCSM servers.
3.1 Molecular docking
The docking scores and binding energies of all chemical constituents of Solanum
indicum targeting antistress activity (PDB ID: 3VW6) and binding interactions with amino
acid residues are presented in Table 1 respectively.

Table 1. Binding interaction of ligands from Solanum indicum targeting antistress activity
(PDB ID:3VW6)

Chemical Docking Score

Sr. No. PubChem ID
constituent 3VW6
1. Solanidine 65727 -9.1
2. Solasodine 5250 -7.4
3. Solavetivone 442399 -6.9
4. Solafuranone 11107208 -8.4
5. Scopoletin 5280460 -6.7
N-p-trans-
6. 5372945 -7.0
Coumaroyltyramine
7. Isofraxidin 5318565 -6.9
8. Lauric acid 3893 -3.7
9. 𝛽-sitosterol 222284 -7.4
10. Diosgenin 99474 -8.5
Standard Drug
11 Fluvoxamine 5324346 -6.6

The binding affinities of phytoconstituents ranged from –9.1 to −3.7 kcal/mol. From the
docked results, it is evident that the compounds, Solanidine exhibit the most favourable binding
affinity (−9.1 kcal/mol) in complex with selected macromolecules (PDB ID: 3VW6) as
compared to other docked compounds i.e., Diosgenin (−8.5 kcal/mol), Solafuranone (−8.4
kcal/mol), Solasodine (−7.4 kcal/mol), Beta sitosterol (-7.4 kcal/mol), N-p-trans-

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Coumaroyltyramine (−7.0 kcal/mol), Isofraxidin (−6.9 kcal/mol), Solavetivone (−6.9

kcal/mol), Scopoletin (−6.7 kcal/mol), Lauric acid (−3.7 kcal/mol). Visual examination of the
computationally docked optimal binding poses of phytoconstituents on selected
macromolecules (i.e., 3VW6) revealed the significant involvement of various types of
interactions, such as hydrogen bonding and hydrophobic interactions, including π–π stacking
and π–alkyl and alkyl interactions, in the stability of the binding of the phytoconstituents to
3VW6.
The binding affinity of the standard (Fluvoxamine) for 3VW6 is -6.6 kcal/mol.
3.1.1. Solafuranone, 3VW6
The number of intermolecular hydrogen bonds, the binding energy of ligand-3VW6 stable
complexes, and the number of nearest amino acid residues were also determined for selected
compound Solafuranone. All synthesized derivatives formed complexes with target proteins.
Analysis of interactions of the 3VW6 protein complex and ligand Solafuranone showed that
the ligand molecule is oriented due to Pi-alkyl interactions with LEU A: 810 amino acid
residues Conventional Hydrogen bond with LYS A: 709 and Pi-Sigma interaction with LEU
A: 686 amino acid residue were found.
3.1.2. Isofraxidin, 3VW6
An analysis of the interactions between the 3VW6 protein complex and the Isofraxidin ligand
was also carried out, which showed that the ligand molecule is oriented due to conventional
hydrogen bond with the amino acid residue LYS A: 688 and SER A: 821 and Pi-Alkyl and
Alkyl interaction with ALA A:707, LUE A:686 and LUE A: 810, Pi-Sigma interaction with
VAL A: 694 amino acid residue were found.
3.1.3. Fluvoxamine, 3VW6
The binding affinity of the standard (Fluvoxamine) for 3VW6 is -6.6 kcal/mol. the interactions
between the 3VW6 protein complex and the Fluvoxamine ligand was also carried out, which
showed that the ligand molecule is oriented due to conventional hydrogen bond with the ASP
A:803, ARG A:767, SER A:821 amino acid residue, Pi-Alkyl and Alkyl interaction with ALA
A:7047, MET A:754, LEU A:810, VAL A:694 amino acid residue and Salt bridge with ASP
A:822 amino acid residue were found.

Table No. 2. Interaction with amino acid residue.

Main amino acid interactions

Binding Pi-alkyl, alkyl, Pi-S/Pi-Pi, Van der
Sr.
Energy T Waals
No Molecule H bond
(kcal/mo shaped/halogen/unfavour interactio
.
l) able donor-donor ns
interactions
No
No
1 Solanidine -9.1 interactio PHE A: 782, TRP A: 770
interaction
n
ASN A:
LEU A: 741, VAL A: 704, No
2 Solasodine -7.4 702, ASP
ARG A: 705 interaction
A: 699

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No
No
3 Solavetivone -6.9 interactio LEU A: 810, VAL A: 694
interaction
n
LYS A: No
4 Solafuranone -8.4 LEU A: 810, LEU A: 686
709 interaction
VAL A:
757,
MET A: 754, LEU A: 810,
GLU A: No
5 Scopoletin -6.7 VAL A: 694, ALA A: 707,
755, interaction
LEU A: 686
LYS
A:709
N-p-trans-
VAL A: GLY A: 759, VAL A: 694, No
6 Coumaroyltyrami -7.0
757 MET A: 754 interaction
ne
LYS A:
ALA A: 707, LEU A: 686, No
7 Isofraxidin -6.9 688, SER
LEU A: 810, VAL A: 694 interaction
A: 821
GLN A: No
8 Lauric acid -3.7 LEU A: 810
756 interaction
LEU A: 686, ALA A: 707,
No
MET A: 754, LEU A: 810, No
9 𝛽-sitosterol -7.4 interactio
VAL A: 738, VAL A: 694, interaction
n
VAL A: 685, LYS A: 688
ASN A: No
10 Diosgenin -8.5 ARG A: 705, VAL A: 704
702 interaction
ASP A:
803, ASP A: 822, VAL A: 694,
No
11 Fluvoxamine -6.6 ARG A: LEU A: 810, MET A: 754,
interaction
767, SER ALA A: 707
A: 821

3.2. ADMET study

Pharmacokinetic profile (ADME) and toxicity predictions of the ligands are important attentive
parameters during the transformation of a molecule into a potent drug. In the present study,
these parameters were assessed using SwissADME and pkCSM. The absorption potential and
lipophilicity are characterized by the partition coefficient (Log P) and topological polar surface
area (TPSA), respectively. For better penetration of a drug molecule into a cell membrane, the
TPSA should be less than 140 Å. However, the value of Log P differs based on the drug target.
The ideal Log P value for various drugs are as follows: oral and intestinal absorption, 1.35 −
1.80; sublingual absorption, > 5; and central nervous system (CNS)11. The aqueous solubility
of ligands ideally ranges from − 6.5 to 0.5 12, while the blood brain barrier (BBB) value ranges
between − 3.0 and 1.2 13. In addition, non-substrate P-glycoprotein causes drug resistance 14.

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In our study, all the selected ligands followed the TPSA parameter, P-glycoprotein non-
inhibition, thereby showing good intestinal absorption and an acceptable range of BBB
values. All the compounds showed aqueous solubility values within the range. Further, it was
predicted that the selected ligands do not show AMES toxicity, hepatotoxicity, and skin
sensitivity. In addition, it did not inhibit hERG-I (low risk of cardiac toxicity). Lipinski’s rule
violations, T. pyriformis toxicity, minnow toxicity, maximum tolerated dose, rat acute oral
toxicity, and chronic toxicity are depicted in table.

Table 3. ADME and toxicity predicted profile of ligands with superior docking score
Aque Huma
Bloo
ous n
d
MW TPS HB Hb solubi intestin
ADMET Log Brai
Formula (g/m A don accep lity al
Properties P n
ol) (Ȧ2) er tor (Log absorp
Barr
mol/L tion
ier
) (%)
C27H43N 397. 5.65 23.47 0.69
Solanidine 1 2 -4.927 92.975
O 647 5 Ȧ2 5
C7H43NO 413. 5.28 41.49 0.03
Solasodine 2 3 -3.809 92.324
2 64 69 Ȧ2 5
218. 3.90 17.07 0.63
Solavetivone C15H22O 0 1 -4.615 95.873
33 42 Ȧ2 5
232. 3.18 26.30 0.20
Solafuranone C15H20O2 0 2 -3.551 95.523
323 764 Ȧ2 6
-
192. 1.50 59.67
Scopoletin C10H8O4 1 4 -2.504 95.277 0.29
17 72 Ȧ2 9
N-p-trans- -
C17H17N 283. 2.46 69.56
Coumaroylty 3 3 -3.165 90.031 0.55
O3 327 99 Ȧ2
ramine 2
-
222. 1.51 68.90
Isofraxidin C11H10O5 1 5 -2.458 95.588 0.37
196 58 Ȧ2 7
200. 3.99 37.30 0.05
Lauric acid C12H24O2 1 1 -4.181 93.379
322 19 Ȧ2 7
414. 8.02 20.23 0.78
𝛽-sitosterol C29H50O 1 1 -6.773 94.464
718 48 Ȧ2 1
414. 5.71 38.69
Diosgenin C27H42O3 1 3 -5.713 96.565 0.2
63 39 Ȧ2

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-
C15H21F3 318. 3.20
Fluvoxamine 56.84 1 4 -3.641 90.686 0.31
N2O2 339 15
Ȧ2 2

Table 3 Continued

Total Max

clearanc tolerate
P-glyco- d
e Bioavailabili AME hERG hERG
ADMET S I II
protein ty score dose
[Log toxici inhibit inhibit
Properties substra
mL/ score ty [Log or or
te
(min.kg mg/
)] (kg.d)]
Solanidine YES 0.028 0.55 NO -0.882 NO YES
Solasodine YES 0.09 0.55 NO -0.375 NO YES
Solavetivone NO 1.225 0.55 NO 0.044 NO NO
Solafuranone NO 1.256 0.55 NO 0.526 NO NO
Scopoletin NO 0.73 0.55 NO 0.614 NO NO
N-p-trans-
Coumaroyltyra YES 0.265 0.55 NO -0.213 NO YES
mine
Isofraxidin NO 0.713 0.55 NO 0.56 NO NO
Lauric acid NO 1.623 0.85 NO -0.34 NO NO
𝛽-sitosterol NO 0.628 0.55 NO -0.621 NO YES
Diosgenin NO 0.328 0.55 NO -0.559 NO YES
Fluvoxamine NO 0.589 0.55 NO 0.213 NO YES

Table 3 Continued
Minn
Oral rat
Acute T.Pyrifo ow
chronic Lipinsk
oral rat Hepat Skin rmis toxici
ADMET toxicity i’s rule
toxicity. otoxici sensat toxicity ty
Properties (Log Violatio
LD50(mo ty ion (Log (Log
mg/kgbw/ n
l/kg) μg/L) mmol
day)
/L)

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-
Solanidine 2.596 1.334 YES NO 0.378 YES (1)
0.493
Solasodine 2.489 1.332 YES NO 0.311 0.381 YES (1)
Solavetivone 1.643 1.19 NO YES 1.453 0.874 YES (0)
Solafuranone 1.865 1.947 NO YES 2.151 0.557 YES (0)
Scopoletin 1.95 1.378 NO NO 0.516 1.614 YES (0)
N-p-trans-
Coumaroylty 2.17 1.271 NO NO 1.008 1.514 YES (0)
ramine
Isofraxidin 2.326 1.825 NO NO 0.431 1.862 YES (0)
-
Lauric acid 1.511 2.89 NO YES 0.954 YES (0)
0.084
-
𝛽-sitosterol 2.552 0.855 NO NO 0.43 YES (1)
1.802
Diosgenin 1.921 1.452 NO NO 0.399 0.247 YES (1)
Fluvoxamine 3.311 1.314 NO NO 1.183 0.841 YES (0)

3.3. Interaction of Standard Drug (Fluvoxamine) with 3VW6

Fig. 2. Docking scores and binding interaction of Fluvoxamine (PDB ID: 3VW6). The
ligand is shown in line and stick representation along with its 2D diagram and hydrogen
bond interaction.

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3.4. Interactions of phytoconstituents with 3VW6

A.Solafuranone

B.Isofraxidin

Fig. 3. Docking scores and binding interaction of phytoconstituents (PDB ID: 3VW6).
The ligand is shown in line and stick representation along with its 2D diagram and
hydrogen bond interaction.

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3.5. Boiled Egg

Fig no. 4. Combined Boiled Egg Diagram

Table no. 4. Name of molecules contained in Boiled Egg Diagram

MOLECULE NUMBER MOLECULE NAME
1 Solanidine
2 Solasodine
3 Solavetivone
4 Solafuranone
5 Scopoletin
6 N-p-trans-Coumaroyltyramine
7 Isofraxidin
8 Lauric acid
9 𝛽-sitosterol
10 Diosgenin
11 Fluvoxamine

BOILED means Brain Or IntestinaL EstimateD permeation predictive model. The boiled egg
diagram shows two regions white region and yellow region.
The white region is the physicochemical space of molecules with highest probability of being
absorbed by the gastrointestinal tract, and the yellow region (yolk) is the physicochemical
space of molecules with highest probability to permeate to the brain.
In addition, the points are coloured in blue if predicted as actively effluxed by P-gp (PGP+)
and in red if predicted as non-substrate of P-gp (PGP-).
4. CONCLUSION
In this study, we have carried out an in-silico screening of the phytoconstituents of Solanum
indicum plant. This study demonstrated the sixteen compounds from Solanum indicum plant,
(Solanidine, Solasodine, Solavetivone, Solafuranone, Scopoletin, N-p-trans-
Coumaroyltyramine, Isofraxidin, Lauric acid, 𝛽-sitosterol, Diosgenin). The selected
phytocompounds showed docking scores ranging from –9.1 to −3.7 kcal/mol in 3VW6. Among
all, Solafuranone and Isofraxidin gave the highest binding energy (−8.4 kcal/mol) and (−6.9

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kcal/mol) in complex with 3VW6, whereas the reference compound, Fluvoxamine showed a
docking score with a binding energy of -6.6 kcal/mol. Furthermore, these ligands exhibited
good ADMET properties. To summarize, phytoconstituents present in Solanum indicum
possess strong effects against 3VW6 and could be further evaluated for their antistress effect,
as well as for the development of alternative drugs with fewer side effects for the treatment of
stress.

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49

Design, Synthesis, Spectral Characterisation and Antibacterial Screening

of Some Novel 4-Substitutedimino-1,3,5-Dithiazine Along With Pyrimidine
Nucleus
S. B. Sarkatea*, S. A. Waghmareb, K. U. Dongarec, R. N. Ingoled
a,b,c-
Department of Chemistry, Ghulam Nabi Azad Arts, Commerce and Science College Barshitakli Dist. Akola
444401 (M.S.), India.
d-
Department of Chemistry, Shri Vitthal Rukhmini College, Sawana,Mahagaon, Dist.Yavatmal,(M.S.) India
*Corresponding author- [email protected]

ABSTRACT
In recent times in the laboratory, synthesis of 2-(2-Phenylimino-4-substitutedimino-
1,3,5-dithiazino) aminopyrimidine (IIIa-e) were synthesized by refluxing 2-(5-phenyl-2,4-
dithiobiureto) pyrimidine (I) with alkyl/arylisocyanodichlorides (IIa-e) in acetone-ethanol
medium in 1:1 molar proportion. The structures of all the synthesized compounds were
acceptable on the basis of chemical characteristics, elemental analysis, spectral studies and
their antibacterial screening against the gram positive and gram negative bacteria such as S.
typhi, E. coli, S. aureus, A. Aerogenes, B. Subtilis and B. Megatherium.etc.
INTRODUCTION
Heterocycles containing organic molecules are more fascinating, because they are
convenient in a variety of applications, a multitude of uses are included by the heterocycles'
size and heteroatom diversity. These compound nitrogen, oxygen and sulphur are composed
of six member1-5, five member6-7 fused heterocycles with aromatic rings. Particularly
noteworthy are the numerous biological and industrial uses for heterocycles that combine
sulphur and nitrogen in one ring. Six member two sulphur and one nitrogen atom prepare 1,3,5-
dithiazine. It is one of the six member heterocycles that functions as a strong medication in the
sectors of medicine, agriculture, and industry9-8.
Tayade10, Pathe11 and Mur12 Synthesized numerous heterocycles containing 1,3,5-
dithiazines as main nucleus. Each 1,3,5-dithiazino moiety has different applications according
to the substituent attached to the basic nucleus of the 1,3,5-dithiazine. It has been also observed
during literature study that, 1,3,5-dithiazino nucleus and its derivatives possesses biological
and medicinal effective properties13-14.
Substituted isocyanodichlorides were used by researchers to synthesise 1,3,5-dithiazines in the
laboratory. This 1,3,5-dithiazine synthesis technique is quicker, less complicated, less
expensive and requires less time.
MATERIAL AND METHOD
Material
All the chemical used were of loba chemie (AR grade).
Method
In the present experiment for the synthesis of different substituted 1,3,5-dithiazino
aminopyrimidine is conventional refluxing under electronic water bath for different hours for
different experiment.
EXPERIMENTAL
All the chemicals used for the synthesis were purified. After refluxing the purity of the
compounds were checked by TLC (aluminium TLC) with thin layer thickness of 200 um. The
melting points of all synthesize compounds will be recorded using hot paraffin bath. The carbon
and hydrogen analysis were carried out on Carlo-Ebra-1106 analyser Nitrogen estimation were
carried out with colmon-N-analyzer-29. IR spectra were recorded with Bruker spectrometer in

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the range 4000-400 cm-1. PMR spectra were recorded on VARIAN 400 MHz spectrometer
with TMS as internal standard using CDCL3 and DMSO Solvent.
GENERAL PROCEDURE
A reaction of 2-(5-phenyl-2,4-dithiobiureto)pyrimidine (I) and substitute
disocyanodichloride (IIa-e) in 1:1 molar ratio refluxed on water bath in acetone-ethanol
medium for 2 hours. The evolution of the hydrochloride gas was clearly observed during
refluxtion. After distillation of excess solvent orange product isolated which on basification
with dilute ammonium hydroxide orange crystalline products obtained.
Similar, procedure was adopted for the synthesis of all the derivatives in the series.
The tentative reaction for the formation of product is depicted below,
Reaction
N NH NH NH Cl 1

N
+ N
R
S S Cl

2-(5-Phenyl-2,4-dithiobiureto)pyrimidine (I) Substitutedisocyanodichloride

(IIa-e)

Reflux 2 hr. Acetone +

ethanol

N
N S
1
N R
S N
NH
N
2-(2-Phenylimino-4-substitutedimino-1,3,5-dithiazino)aminopyrimidine

(IIIa-e)

Similarly, 2-(5-phenyl-2,4-dithiobiureto)pyrimidine (I) were react with phenylisocyanochloride

(IIa), ethylisocyanodichloride (IIb), tertbutylisocyanodichloride (IIc), P-tolylisocyanodichloride (IId)
and 4-chlorophenhylisocyanodichloride (IIe) by the above mentioned method to isolate 2-[2,4-
di(phenylimino)-1,3,5-dithiazino] aminopyrimidine (IIIa), 2-(2-phenylimino-4-ethylimino-1,3,5-
dithiazino) aminopyrimidine(IIIb), 2-(2-phenylimino-4-tert-butylimino-1,3,5-dithiazino)
aminopyrimidine (IIIc), 2-[2-phenylimino-4-(4-methylphenylimino)-1,3,5-dithiazino]aminopyrimidine
(IIId) and 2-[2-phenylimin-4-(4-chlorophenylimino)-1,3,5-dithiazino]aminopyrimidine (IIIe).
RESULT AND DISCUSSION
Spectral characterization results for all the synthesized compounds are given below
Spectral Characterization
1. 2-(2,4-diphenylimino-1,3,5-dithiazino)aminopyrimidine (IIIa)
Colour-Yellow solid, Molecular formula- C15H14N6S2, Yield 85%, M.P. 1620C, %
Composition found (calculated) C-52.90 , H-4.83 , N-25.20, S-23.21 , FTIR (Kbr) vcm-
3245.22 N-H stretching, 3051.47 (C-H Ar Stretching), 3187.38 (N-H Amido), 1950.37 (C-H
Ar Bending,), 1185.99 (C=S Stretching), 670.04(=C-H bending); 1H NMR (400MHz CDCL3
δ ppm), 8.4ppm (2H, double, CH) of pyrimidine, 7.2ppm (2H, CH, doublet) of pyrimidine, 7.2

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ppm ( 2H, , doublet CH) of phenyl, 3.4 ppm (1H, singlet NH), 4.6 ppm 1H singlet, 7.39 1H
triplet CH benzene, 1H triplet CH benzene 2.2 ppm 2H quartet, 2.3 ppm 3H triplet Mass m/z
390.30
2. 2-(2-phenylimino-4-ethylimino-1,3,5-dithiazino)aminopyrimidine (IIIb)
Colour-Yellow solid, Molecular formula- C15H14N6S2, Yield 86%, M.P. 1700C, %
Composition found (calculated) C-52.90 , H-4.83 , N-25.20, S-23.21 , FTIR (Kbr) vcm-
3245.22 N-H stretching, 3051.47 (C-H Ar Stretching), 3187.38 (N-H Amido), 1950.37 (C-H
Ar Bending,), 1185.99 (C=S Stretching), 670.04(=C-H bending); 1H NMR (400MHz CDCL3
δ ppm), 8.4ppm (2H, double, CH) of pyrimidine, 7.2ppm (2H, CH, doublet), 7.2 ppm ( 2H, ,
doublet CH) of phenyl, 3.4 ppm (1H, singlet NH), 4.6 ppm 1H singlet, 7.39 1H triplet CH
benzene, 1H triplet CH benzene 2.2 ppm 2H quartet, 2.3 ppm 3H triplet Mass m/z 342.30
3. 2-(2-phenylimino-4-tertbutylimino-1,3,5-dithiazino)aminopyrimidine (IIIc)
Colour-Yellow solid, Molecular formula- C17H18N6S2, Yield 90%, M.P. 1580C, %
Composition found (calculated) C-55.90 , H-4.83 , N-23.20, S-17.21 , FTIR (Kbr) vcm-
3245.22 N-H stretching, 3045.47 (C-H Ar Stretching), 3140.38 (N-H Amido), 1995.37 (C-H
Ar Bending,), 1164.99 (C=S Stretching), 710.04(=C-H bending); 1H NMR (400MHz CDCL3
δ ppm), 8.2ppm (2H, double, CH), 7.2ppm (2H, CH, doublet), 7.1ppm ( 2H, , doublet CH),
3.5 ppm (1H, singlet NH), 4.5 ppm 1H singlet, 7.40 1H triplet CH benzene, 2.4 9H singlet
Mass m/z 370.50
4. 2-(2-phenylimino-4-tolylimino-1,3,5-dithiazino)aminopyrimidine (IIId)
Colour-Yellow solid, Molecular formula- C20H16N6S2, Yield 82%, M.P. 1670C, %
Composition found (calculated) C-55.90 , H-4.83 , N-23.20, S-15.21 , FTIR (Kbr) vcm-
3200.22 N-H stretching, 2901.47 (C-H Ar Stretching), 316038 (N-H Amido), 1960.37 (C-H
Ar Bending,), 1167.99 (C=S Stretching), 745.04(=C-H bending); 1H NMR (400MHz CDCL3
δ ppm), 8.2 ppm (2H, double, CH) of pyrimidine, 7.3ppm (2H, CH, doublet), 7.2 ppm ( 2H, ,
doublet CH), 3.5 ppm (1H, singlet NH), 4.7 ppm 1H singlet, 7.39 1H triplet CH benzene, 3.4
ppm 3H singlet. Mass m/z 404.60
5. 2-(2-phenylimino-4-p-chlorophenylimino-1,3,5-dithiazino)aminopyrimidine (IIIe)
Colour-Yellow solid, Molecular formula- C19H13N6S2, Yield 80%, M.P. 1450C, %
Composition found (calculated) C-53.90 , H-3.83 , N-20.20, S-15.21Cl-8.45 , FTIR (Kbr)
vcm- 3263.22 N-H stretching, 3051.47 (C-H Ar Stretching), 3216.38 (N-H Amido), 1966.37
(C-H Ar Bending,), 1165.99 (C=S Stretching), 720.04(=C-H bending); 1H NMR (400MHz
CDCL3 δ ppm), 8.4 ppm (2H, double, CH) of pyrimidine, 7.03ppm (2H, CH, doublet), 7.4
ppm ( 2H, , doublet CH), 3.5 ppm (1H, singlet NH), 4.6 ppm 1H singlet, 7.39 1H triplet CH
benzene, Mass m/z 424.80.

CONCLUSION
In the present work is cheaper and less time consuming method for synthesis of organic
compound (IIIa-e). In all the synthesized compounds give the maximum yield of product (III
a-e). A variety of pyrimidine based 1,3,5-dithiazine derivative can be synthesized for their
antimicrobial activities adopting the method.

PHARMACOLOGICAL STUDIES
Antimicrobial activity
All the synthesized compounds (III-a) to (III-e) were screened for antibacterial activity
against S. typhi, E. coli, S. aureus, A. Aerogenes, B. Subtilis and B. Megatherium. by disc
diffusion method was performed using mueller hinton agar as well as nutrient agar medium.
Each and every compound was tested at conc. 50 μg/ml in ethanol. The zone of inhibition of
all the synthesized compounds were measured after 24 hour incubation at 37 0C. Standard drug
used for comparison the activity was Ciprofloxacin.

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Table 1: Synthesized compound IIIa-e activity against Gram +ve and Gram -ve bacteria

Table No. 1.1 - Antibacterial activity of synthesized compound against bacteria (Zone
of inhibition in mm) (after 24 hrs at 37 oC temp)

Gram positive Gram negative

SR B.
Compd. S. B. S. E. A.
No. megatheriu
aureus subtilis typhi coli aerogenes
m
1 SBS-IIIa 18 12 16 14 18 11
2 SBS-IIIb 11 12 14 14 11 12
3 SBS-IIIc 11 11 12 11 11 11
4 SBS-IIId 10 12 12 12 11 11
5 SBS-IIIe 14 12 14 12 16 12
Std
6 18 16 20 18 20 14
Ciprofloxacin

REFERANCE

1. Zhang, D. H., Zhang, Z., & Shi, M. (2012). Transition metal-catalyzed carbocyclization of
nitrogen and oxygen-tethered 1, n-enynes and diynes: synthesis of five or six-membered
heterocyclic compounds. Chemical Communications, 48(83), 10271-10279.
2. Larrosa, I., Romea, P., & Urpí, F. (2008). Synthesis of six-membered oxygenated heterocycles
through carbon-oxygen bond-forming reactions. Tetrahedron, 64(12), 2683-2724.
3. Sanu, M. C., Joseph, J., Chacko, D., Vinod, B., & Daisy P., A. (2021). Review on six membered
nitrogen containing heterocyclic compounds with various biological activities. International
Journal of Pharmaceutical Sciences Review and Research, 69(2), 64-68.
4. Walton, J. C. (2016). Synthetic strategies for 5-and 6-membered ring azaheterocycles facilitated
by iminyl radicals. Molecules, 21(5), 660.
5. Huh, D. N., Cheng, Y., Frye, C. W., Egger, D. T., & Tonks, I. A. (2021). Multicomponent
syntheses of 5-and 6-membered aromatic heterocycles using group 4–8 transition metal
catalysts. Chemical Science, 12(28), 9574-9590.
6. Sergey P., Vladimir N., Andrey, E., & A. Pimerzin, E. Vishnevskaya E., Thermodynamic
analysis of strain in the five-membered oxygen and nitrogen heterocyclic compounds. The
Journal of Physical Chemistry, 115( 10), 1992–2004.
7. Kaur, N., Yadav, N., & Verma, Y. (2023). Acetamidine in heterocycle synthesis. Synthetic
Communications, 53(9), 577-614.
8. Gujjar, K. N., & Narasimha S M., (2023).A Review: Important applications of Heterocyclic
Compounds. Europeam Chemical Bulletin, 12(12), 625-630.
9. Sharma, P. K., Amin, A., & Kumar, M. (2020). A review: Medicinally important nitrogen
sulphur containing heterocycles. The Open Medicinal Chemistry Journal, 14(1), 49-64.
10. Tayade, D. T., & Padhen, S. S. (2016). Synthesis and Characterization of 1-Phenyl-3- [4-(2-
Substitutedimino-4-Substitutedimino-1,3,5-Dithiazino) Aminophenyl]- Prop-2-Ene-1-Ones.
International Journal of Pharmacy and Pharmaceutical Research, 7 (1), 53-58.
11. Pathe, P. P., & Paranjpe, M. G. (1984). Preparation of 5‐aryl‐4‐ arylimino‐6‐benzylimino‐2‐
phenylimino‐1,3,5‐dithiazine. Indian chemical society, 15(43), 149-150.
12. Mur, V. I. (1964). 2, 4, 6-trichloro-1, 3, 5-triazine (cyanuryl chloride) and its future
applications. Russian Chemical Reviews, 33(2), 92-103.
13. Panpaliya, K. S., Tayade, D. T., Shaikh, R. S., & Thakare, A. N. (2017). Synthesis of 1-phenyl-
3-substituted- 2,6-dithio-4- amino-[(2-phenylthiocarbamido)- 1,3-benzothiazolo] -1,3,5-triazine
and their effects on germination pattern of sorghum vulgare. Online International
Interdisciplinary Research Journal, 1, 6-9.
14. Deohate, P. P., & Berad, B. N. (2005). Synthesis and antimicrobial activity of 1, 3, 5-thiadiazines
and their isomerism into 1, 3, 5-triazines. Indian journal of Chemistry, 44B, 638-642.

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50

Comparative Account of Partial Molar Volumes and Compressibilities of

Aqueous-(L-Arginine + Glucose/Lactose) Solutions at 310.15 K
R. V. Dudhate1, H. N. Pawar1, B. R. Bhosle 1, S. D. Deosarkar1, S. D. Deshmukh2, and
P. S. Bodkhe3
1
School of Chemical Sciences, Swami Ramanand Teerth Marathwada University, Nanded
2
College of Engineering and Technology, Akola
3
Vidya Bharati Mahavidyalaya, Camp Amravati

Abstract: Partial molar volumes and compressibilities of aqueous-(L-Arginine + Glucose/Lactose)

solutions have been determined from measured densities and ultrasonic velocities at 310.15 K. The
measured, calculated and graphically determined properties viz. densities and ultrasonic velocities,
apparent molar volumes and compressibilities and partial molar volumes and compressibilities and have
been interpreted in terms of different interactions in studied solutions. Effect of Glucose/Lactose on
structural orientation, hydration behavior and different interactions including hydrogen bonding has
been evaluated. Study has importance in amino acid-sugar interactions.

Introduction:
L-Arginine is an essential basic amino acid found in protein rich food and strongly interacts with organic
acids as well as carbohydrates through hydrogen bonding and hydrophilic/hydrophilic interactions.
Glucose is a most abundant monosaccharide sugar [1-2]. Lactose (milk sugar) is a disaccharide sugar
used in the food industry [3].
Numbers of researchers have carried out molecular interaction studies of L-Arginine in aqueous and
various cosolutes including drug [4-8] solutions. These interactions lead to the perturbation of structure
and orientation of aqueous L-Arginine. Sugar interactions with amino acid specifically L-Arginine
through volumetric and ultrasonic studies in solution is lacking. In view of importance of these studies
in this short communication, we report comparison of partial molar volumes and compressibilities of
these solutions at 310.15 K.

Experimental:
L-Arginine (Himedia, 174.2 g/mol) solutions in water and aqueous Glucose (Himedia, 180.16
g/mol)/Lactose (Himedia, 342.30 g/mol) were prepared in double distilled water. Details of
experimental work (measurement of densities and ultrasonic velocities) are explained in our previous
papers [9-11]. Apparent and partial molar volume and compressibility and corresponding transfer
properties are determined using standard relations [12-17].

Results and discussion:

It is seen from Figure 1 and 2 that the measured densities (  ) and ultrasonic velocities u increase with
increase in the concentration of L-Arginine as well as increase in the concentration of cosolutes (Figure
1 A and C, 2 A and C). The apparent molar volumes ( V 2, ) and compressibilities (  2, ) increase with
increase in concentration L-Arginine as well as increase in cosolute concentration (Figure 1 B and C, 2
B and D). However these values are larger in Lactose solution compared to Glucose solution. This is
attributed to the more number of –OH groups in Lactose compared to Glucose which makes loose
structure, expansion in volume of hydration with more compression in Lactose.
o
The partial molar properties, partial molar volumes ( V2, ) and partial molar compressibilities (  S0, 2 , )
are determined from straight line equations as an intercept of lines using Masson’s and Gucker’s
equations and solute-solute interaction parameters ( S v and S k ) are determined as a slope of these lines
and are listed in Table 1 and 2. Partial molar properties and transfer properties ( tV2o, and t S0, 2 , ) of
L-Arginine increases on addition of cosolutes and further increases with their concentration. This is due

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to the overlap of cospheres of solute and cosolute and release of water molecules into the bulk solution
(Scheme 1).
A
B

1002 129
129
1000 128
128
Density

AMV
998 127
127
996
126
994 126
125
992 125
0 0.05 0.1 0.15 0.1 0.2 0.3 0.4
Concentration Concentration
Water 0.02 0.05 0.1 Water 0.02 0.05 0.1

D
C 0.1 0.2 0.3 0.4
1575
1570 -12.60
Ultrasonic vvelocity

1565 -12.80
1560 -13.00
1555
-13.20
1550
AMC

1545 -13.40
1540 -13.60
1535 -13.80
1530
-14.00
0 0.05 0.1 0.15
-14.20
Concentration
Water 0.02 Concentration
0.05 0.1 Water 0.02 0.05 0.1

Figure 1 A-D. The,  V 2, and u  2, values of (Aq. L-Arginine + Glucose) at 310.15 K

A B
1002 131
130
1000
129
Density

998
AMV

128
996 127
994 126
992 125
0 0.05 0.1 0.15 124
Concentration 0.1 0.2 0.3 0.4
Concentration
Water 0.02 0.05 0.1 Water 0.02 0.05 0.1

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C D
1570 0.1 0.2 0.3 0.4
1565
Ultrasonic velocity
-11.00
1560
-11.50
1555
-12.00
1550
-12.50

AMC
1545
1540 -13.00
1535 -13.50
1530 -14.00
0 0.05 0.1 0.15 -14.50
Concentration Concentration
Water 0.02 0.05 0.1 Water 0.02 0.05 0.1

Figure 2 A-D. The,  V 2, and u  2, values of (Aq. L-Arginine + Lactose) at 310.15 K

o
Table 1. The V2, , S v , tV2o, and  S0, 2, , t S0, 2 , values of (Aq. L-Arginine + Glucose) at
310.15 K
System/property V2o, Sv tV2o,

Water 124.56 3.307 0.000

0.02M aq. Glucose 124.73 5.267 0.170
0.05M aq. Glucose 125.73 4.954 1.170
0.10 M aq. Glucose 126.69 4.894 2.130
System  S0, 2 , Sk t S0, 2 ,
Water -14.190 1.361 0.000
0.02M aq. Glucose -13.797 0.781 0.393
0.05M aq. Glucose -13.256 0.146 0.934
0.10 M aq. Glucose -12.908 0.426 1.282

o
Table 2. The V2, , S v , tV2o, and  S0, 2, , t S0, 2 , values of (Aq. L-Arginine + Glucose) at
310.15 K
System/property V2o, Sv tV2o,

Water 124.56 3.307 0.000

0.02M aq. Lactose 125.40 4.624 0.840
0.05M aq. Lactose 126.91 3.769 2.350
0.10 M aq. Lactose 128.05 5.801 3.490
 S0, 2 , Sk t S0, 2 ,
Water -14.190 1.361 0.000
0.02M aq. Lactose -13.318 1.643 0.872
0.05M aq. Lactose -12.596 1.687 1.594
0.10 M aq. Lactose -11.873 1.047 2.317
o
Foot Note: V2, =cm3·mol-1, S v =cm3·mol-3/2·kg1/2.  S0, 2, =×10-14 m3·mol-1·Pa-1, S k =×10
-14

m3∙mol-3/2∙kg1/2 ∙Pa-1, tV2o, =cm3·mol-1,  S , 2, =×10-14 m3·mol-1·Pa-1.

0

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OH
HO OH
HO OH
H
H O HO H H O
O H OH H
H HOH HOH
O H H H AcH H H
H O OH H OH
H O
ClOH H
H H H HO
O O O
H
H
O O HO OH
H H NH2 H OH
H NH2 H OH H H
H2N N OH HO HO H2N N OH H O OH
O H OH O
HO H O NH O O
NH O H HO H
OH
H H H HO H
HO OH O OH
HO H H HO H
H OH O OH
HO H H H H
H HOH HOH
H
OH H
H OH OH H
H HO H O
A OH
HO OH
HO OH

OH
HO
O
H
H O HO H H HO OH
O H OH H HO O O
H HOH HOH HO
O H H H OH HO H H OH
H O OH H OH OH O O
H HO O OH
H OH H
H H H H HO OHO
O O O O HO OH
H H H O H
H NH2 H H H2N H NH2 OH
H2N N OH HO N OH H
O H O
HO HO NH O H HO NH O H
H H H OH H
HO OH O HO O OH
HO H H HO H
H OH O OH
HO H H H HO O H HO
H HOH OH OH HOH
H
OH H O OH
H OH OH H O O
H HO H OH OH
HO
B
OH OH OHO
HO OH
OH

Scheme 1. Hydration of L-Arginine in presence of A) Glucose and B) Lactose

The study has implications in protein-sugar interactions in solution which discloses size of
hydration sphere, specific interactions and solution behavior of amino acids in specific and
proteins in general. The observations in present systems reveals that the hydrogen bonding and
hydrophilic-hydrophilic interactions occurs in L-Arginine and Glucose//Lactose with
dominance of these interactions in Lactose solution.
Conclusion:
Hydration structure of L-Arginine affects in presence of cosolutes glucose and Lactose as a
result of hydrogen bonding and other interactions in solution. The cosolute causes increase in
the volume and compression and weakening of solute-solvent interactions as a cosphere
overlap. This effect is dominant in Lactose solutions.
References:
1. Domb, Abraham J.; Kost, Joseph; Wiseman, David (1998-02-04). CRC Press. 275. ISBN 978-1-4200-4936-
7.
2. Kamide, Kenji (2005). Amsterdam: Elsevier. ISBN 9780080454443.
3. Gerrit M. Westhoff; Ben F.M. Kuster; Michiel C. Heslinga; Hendrik Pluim; Marinus Verhage (2014).
ISBN 978-3-527-30673-2.
4. Anil Kumar Nain, J. Mol. Liq. 113736 (2020).
5. V. D. Umare, Int. J. Res. Appl. Sci. Engg. Tech., 8 (2020).
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52(3):155–161 (2014).
7. R. Palani, S. Balakrishnan and G. Arumugam, J. Phy. Sci. 22(1), 131–141, (2011).
8. Sonu R. Dhumane, Nita S. Ramteke and K. C. Patil, Int. J. Res. Bio. Agri. Tech. 38–41 (2016).
9. Deosarkar, S.D., Arsule, A.D., Kalyankar, T.M. Coll. Surf. A 613, 126052 (2021).
10. Arsule, A.D., Sawale, R.T., Kalyankar, T.M., Deosarkar, S.D., J. Solut. Chem. 49(1), 83–99 (2020).
11. A.D. Arsule, R.T. Sawale, S.D. Deosarkar, J. Mol. Liq. 266 413–424 (2018).
12. Ivanov, E.V., Lebedeva, E.Y., J. Mol. Liq. 222, 1164–1171 (2016).
13. Chauhan, S., Chauhan, M.S., Jyoti, J., Rajni, J. Mol. Liq. 148, 24–28 (2009).
14. Kumar, K., Chauhan, S., Thermochim. Acta 606, 12–24 (2015).
15. Pal, A., Chauhan, N.: J. Mol. Liq. 162, 38–44 (2011).
16. Kumar, K., Chauhan, S., Thermochim. Acta 606, 12–24 (2015).
17. Pal, A., Chauhan, N., J. Mol. Liq. 162, 38–44 (2011).

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51
Studies in the Synthesis of 5-(3-Substitutedthiocarbamido) Aminoindole by
Microwave Technique
M. B. Shahakar1*, R.D. Isankar1, P.V. Raut2
1
Department of Chemistry, Government Vidarbha Institute of Science and Humanities Amravati, MH-444604,
India.
2
Department of Chemistry, Radhabai Sarda Arts, Commerce and Science College, Anjangaon Surji, Amravati,
MH-444705,India.
*Corresponding author E-mail [email protected]

Abstract: We reported synthesis of 5-(3-thiocarbamido)aminoindol, 5-(3-allylthiocarbamido)

aminoindole, 5-(3-o-nitrophenylthio- carbamido)aminoindol, 5-(3-p-
nitrophenylthiocarbamido)aminoindol, 5-(3-m-chloro- phenylthiocarbamido)aminoindole by
interacting 5-chloroindol with thiourea, allyl thiourea, o-nitrophenylthiourea, m-
chlorophenylthiourea respectively. Ethanol was taken and kept in microwave oven and
irradiation was carried out for 2 minutes at 50 °C. The justification and determination of
structures of synthesized molecules were done on the basis of elemental analysis, chemical
characterizations, IR, H1-NMR, C13NMR studies.This green synthetic method increases yield
of products by maintaining the purity of the products this is the novelty of the research work.

Key words: 5-chloroindole, substituted thiourea, green synthesis, microwave technique.

INTRODUCTION

Indole nucleus containing heterocycles created their own identity due to these
applications in life, pharmaceutical, medical, and agricultural sciences. Indoles showed
anticancer, anti-inflammatory, antioxidant activities1-3.

Indole based heterocycles are well known recognised for their antimicrobial, anti-
micotic and inhibition of chemical mediator’s release4, anti-inflammatory5-7, anti-asthama8,
anti-vascular9, antifungal10, activities.At the same time thiocarbamido nucleus showed
antioxidant, anticancer, anti-inflammatory antitubercular activities. Majority of alkaloids
contain indole nucleus which exhibits excellent biological activities i.e.antiviral, antitumor,
antiplasmodial, antibacterial and neurochemicals melatonin and serotonin are more complex
bisindole natural products vincristine and vinblastine these are the key structural component of
many effective synthetic pharmaceuticals4-5.Amino acid tryptophans, the biogenic amines
serotonin, tryptamine and the mammalian hormone melatonin are broadly known indole
derivatives play an significant role in lively activity of humans and animals11.

Synthetic applications of cynoguanidine and 1, 3- diformamido-thiocarbamide had

been briefly explored12. Synthesis of nitrogen, sulphur and nitrogen and sulphur containing
heteroacycles and heterocycles the interactions of cynoguanidine with various thiourea and
isothiocynates have been investigated in sufficient details 13-18. Some of these compounds
showed remarkable pharmaceutical and biological activities19-20. As a wider programme of this
research laboratory we have decided to synthesize a novel series of 5-(3-substituted
thiocarbamido) aminoindole by interacting 5-chloroindole (I) with thiourea (IIa), allyl thiourea
(IIb), o-nitro-phenyl thiourea (IIc), p-nitro-phenyl thiourea (IId), m-chloro-phenyl thiourea(IIe)
in presence of ethanol by using microwave technique. Structure determination and

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identification of the synthesized product is done on the basis of chemical characteristics,

elemental analysis and spectral data.
GENERAL REACTION:-
S
H2N
Cl S
MWI NH

H2N NHR Ethanol NH

NH
5 - Chloro Indole Substituted thiourea
5-(3 -Substitutedthiocarbamido) aminoIndole
(I) (II)
[(IIIa-e)]

Where R= -H, -allyl, -o-nitrophenyl, -p-nitrophenyl, m-chlorophenyl

1. EXPERIMENTAL
The melting point of the allsynthesized compounds was recorded using hot paraffin
bath. The carbon and hydrogen analysis were carried out on Carlo-Ebra 1106 analyzer.
Nitrogen estimation was carried out on Colman-N-analizer-29. IR spectra were recorded on
Perkin Elmer Spectrometer in range 4000-400cm-1 in KBr pellets. The purity of compound was
checked by TLC plate. All chemicals used were of AR-grade.
2.1 5-(3-Thiocarbamido) Amino Indole (IIIa)
A mixture of 5-chloroindole (I) (0.1M, 0.392 gms) and thiourea (0.1M, 0.152 gms) in
ethanol was kept in microwave used to synthesis 5-(3-thiocarbamido) amino indole (IIIa) (1
gm, 97.35%), melting point 1600C.(D)
RESULTS AND DISCUSSION
Properties of (IIIa): It is pale yellow colour crystalline solid having melting point 1600c (D).
It gave positive test for nitrogen and sulphur. Desulphurised with alkaline plumbite solution. It
forms picrate, melting point 1200c. Elemental analysis C {(found 67.58%) calculated 68.30%},
H {(found 5.17%) calculated 5.33%}, N {(found 14.86%) calculated 14.94 %}, S {(found
11.24%) calculated 11.38%}.
IR Spectra
FTIR Spectrum was recorded using KBr-pellets and is reproduced. Important absorption can
be correlated as (cm-1): N-H stretching at 3387.00cm-1 (3500-3000 cm-1), Ar-H stretching at
2690.70cm-1 (3000-2500cm-1), Ar-c=c stretching at 1612.49cm-1 (1600-1500cm-1), C-N
stretching at 1085.92cm-1 (1200-1000cm)-1, N-C=S stretching at 1462.04cm-1 (1550-1250cm-
1
), NN>C=S stretching at 1400.89cm-1 (1400-1200cm-1).
NMR
This spectrum distinctly displayed signals at (δ ppm): signal at 7.2646-7.1726 ppm is due to
Ar-H, signal at 4.2911-4.4564 ppm is due to N-H in aromatic ring, signal at 3.1402-2.2160
ppm is due to NH2 and -NH protons. 13C Spectrum: This spectrum distinctly displayed signals
due to C=S carbon at  181.76 ppm, Ar-C carbon at  155.58-112.58 ppm, -CH2 carbons at 
79.04-78.38 ppm, -CH3 carbons at  4012-38.87 ppm.

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2.2 5-(3-Allylthiocarbamido) Amino Indole (IIIb)

A mixture of 5-chloroindole (I) (0.1 M, 1.56 gm) and allyl thiourea (IIb) (0.1 M, 0.68
gm), in ethanolwas kept in microwave used to synthesis 5-(3-allylthiocarbamido) amino indole
(IIIb) (2 gm, 90%), melting point 1860C.(D)
RESULTS AND DISCUSSION
Properties of (IIIb): It is lemon yellow colour crystalline solid having melting point 1860c
(D). It gave positive test for nitrogen and sulfur. Desulphurized with alkaline plumbite solution.
It forms picrate, melting point 1100c. Elemental analysis C {(found 68.58%) calculated
69.27%}, H {(found 5.11%) calculated 5.35%}, N {(found 15.86%) calculated 16.64 %}, S
{(found 11.34%) calculated 11.48%}.
IR Spectra
The IR spectra was carried out in KBr pellets and the important absorptions can be correlated
as, (cm-1) 3312 (N-H stretching), 1197 (C-N stretching), 3012 (Ar-H stretching), 1736 (N=C-
N stretching), 1512 (Ar-C=C), 1487 (N-C=S) Bending.
NMR
Ar-H protons (6.8720-6.8991 ppm), N-H protons (5.0417-5.0471 ppm), N-H of five membered
ring (4.2921-4.5564 ppm), -CH3 protons (3.4386-3.8638 ppm).
2.3 5-(3-o-nitro-phenyl thiocarbamido) Amino Indole (IIIc)
A mixture of 5-chloroindole (I) (0.1 M, 1.17 gm) and o-nitro-phenyl thiourea (IIc) (0.1 M,
1.212 gm), in ethanol was used to synthesis 5-(3-o-nitro-phenylthiocarbamido) amino indole
(IIIc) (3 gm, 94%), melting point 1920C.(D)
RESULTS AND DISCUSSION
Properties of (IIIc)-It is orange colour crystalline solid having melting point 1920c (D). It gave
positive test for nitrogen and sulphur. Desulphurisd with alkaline plumbite solution. It forms
picrate, melting point 1200c. Elemental analysis C {(found 63.58%) calculated 64.27%}, H
{(found 5.91%) calculated 6.35%}, N {(found 15.06%) calculated 16.64 %}, S {(found
11.34%) calculated 11.48%}.
IR Spectra
The IR spectra was carried out in KBr pellets and the important absorptions can be correlated
as, (cm-1) 3312 (N-H stretching), 1197 (C-N stretching), 3012 (Ar-H stretching), 1736 (N=C-
N stretching), 1512 (Ar-C=C), 1487 (N-C=S) Bending.
NMR
Ar-H protons (6.8720-6.8291 ppm), N-H protons (5.0417-5.0471 ppm), N-H of five membered
ring (4.2920-4.5565 ppm).
2.4 5-(3-p-nitro-phenyl thiocarbamido) Amino Indole (IIId)

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A mixture of 5-chloroindole (I) (0.1 M, 1.16 gm) and o-nitro-phenyl thiourea (IId) (0.1 M, 1.26
gm), in ethanol was used to synthesis 5-(3-p-nitro-phenylthiocarbamido) amino indole (IIId)
(2.9 gm, 94%), melting point 1940C.(D)

RESULTS AND DISCUSSION

Properties of (IIId)-It is pale orange colour crystalline solid having melting point 1940c (D).
It gave positive test for nitrogen and sulphur. Desulphurisd with alkaline plumbite solution. It
forms picrate, melting point 1300c. Elemental analysis C {(found 63.98%) calculated 64.57%},
H {(found 4.91%) calculated 5.59%}, N {(found 15.26%) calculated 16.97 %}, S {(found
11.34%) calculated 11.78%}.
IR Spectra
The IR spectra was carried out in KBr pellets and the important absorptions can be correlated
as, (cm-1) 3612 (N-H stretching), 1097 (C-N stretching), 3092 (Ar-H stretching), 1796 (N=C-
N stretching), 1572 (Ar-C=C), 1447 (N-C=S) Bending.
NMR
Ar-H protons (6.8420-6.8491 ppm), N-H protons (5.0417-5.0471 ppm), N-H of five membered
ring (4.2960-4.5566 ppm).
2.5 5-(3-m-chloro-phenyl thiocarbamido) Amino Indole (IIIe)
A mixture of 5-chloroindole (I) (0.1 M, 1.36 gm) and m-chloro-phenyl thiourea (IIe) (0.1 M,
1.09 gm) in ethanol was used to synthesis 5-(3-m-chloro-phenylthiocarbamido) amino indole
(IIIe) (2.4 gm, 95%), melting point 1930C.(D)

RESULTS AND DISCUSSION

Properties of (IIIe)-It is ivory colour crystalline solid having melting point 1930c (D). It gave
positive test for nitrogen and sulphur. Desulphurisd with alkaline plumbite solution. It forms
picrate, melting point 1800c. Elemental analysis C {(found 62.98%) calculated 63.57%}, H
{(found 4.31%) calculated 5.89%}, N {(found 14.26%) calculated 15.97 %}, S {(found
11.24%) calculated 11.94%}.
IR Spectra
The IR spectra was carried out in KBr pellets and the important absorptions can be correlated
as, (cm-1) 3412 (N-H stretching), 1037 (C-N stretching), 3192 (Ar-H stretching), 1996 (N=C-
N stretching), 1672 (Ar-C=C), 1457 (N-C=S) Bending.
NMR
Ar-H protons (6.9420-6.8491 ppm), N-H protons (5.0417-5.0461 ppm), N-H of five membered
ring (4.2760-4.5596 ppm).

ACKNOWLEDGMENTS
Author is very much thankful to Dr. R. D. Isankar, Assistant Professor dept. of
chemistry Govt. Vidarbha Institute of Science and Humanity Amravati, Also thankful to Dr.

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D. T. Tayade, professor dept. of chemistry Govt. Vidarbha Institute of science and Humanity
Amravati, for valuable help during this work.
References
1. R. D. Isankar, D. T. Tayade, Journal of medicinal chemistry and drug discovery, 02, 2017,
541-545.
2. P.S. Bodkhe A.B. Wadekar, R.D. Isankar and D.T. Tayade, Journal of chemistry and
chemical sciences, 9(2), 2019, 45-48.
3. K. D. Tayade, D. A.Pund, R.D.Isankar, S.U.Patil, Journal of Medicinal chemistry and Drug
discovery, 1, 2016, 15-19.
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5. Radwan M.A. Ragab E.A., Sabri N.M., El-Shenaw., Bioorg. Med. Chem. Lett., 15, 2007,
3832-3841.
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Chem., 14, 2000, 17-33.
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8. Stanton J.L., Ackerman M.H. J. Med. Chem., 26, 1983, 986-989.
9. Ryu C.K., Lee J.Y., Jeong S.H., Nho J.H., Bioorg. Med. Chem. Lett., 19. 2009, 146-148.
10. Archana, Sachin Saini, Synthesis and anticonvulsant studies, Drug Res, 69 (08), 2019,
445- 450.
11. Kumar and Ritika, Future Journal of pharmaceutical sciences, 6, 2020, 121.
12. Syed Muhammad Umer, Mehwish Solangi, Khalid Mohammad Khan, Molecules, 27,
2022, 7586.
13. Richard P. Nugent, Scot Pounds, Crist N. Filer, Applied Radiation and Isotopes, 69, 2011,
423-425.
14. BugaenkoDmiry I., Karchava Alexander V., Yurovskaya Marina A., Russ. Chem., 88 (2),
2019, 99-159.
15. Tayade D. T. Ph.D Thesis Amravati University,Amravati 1996.
16. Tayade D.T., Raghuwanshi M.R., Bhagwatkar R.A., International Journal of Chemistry
Canada, 3(2), 2011, 74-78.
17. Bhagwatkar R.A., Tayade D.T., Orbital Elec. J.Chem., Campo Grande Brazil, 3(1), 2011,
53-56.
18. Tayade D.T., Bhagwatkar R.A, Panpalia R.C., International Journal of Chemistry Canada,
2(2), 2010, 41-43.
19. Tayade D.T., Pund D.A., Bhagwatkar R.A., Rathod D.B., Bhagwatkar N.A., International
Journal of Chemistry Canada, 3(1), 2011, 36-41.
20. Raut P. V., Waghmare S. A., Tayade D. T., JETIR7(2), 2020, 437-439.

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52
Understanding the Role of Lipophilicity and Pharmacophore Elements in
Pyrimidinedione based BCAT1 Inhibitory Activity
S. R. Deshmukha, P.P. Nalawade*b, V. H. Masandb , S. D. Thakurc, P. S. Navaleb
a
Department of Chemistry, PRMIT&R College, Badnera-Amravati, Maharashtra, India
b
Department of Chemistry, Vidya Bharati Mahavidyalaya, Amravati, Maharashtra, India
c
Department of Chemistry, RDIK and NKD College, Badenera-Amravati, Maharashtra, India
Abstract:
This paper presents the findings from pharmacophore-oriented studies on BCAT1 inhibitors.
Understanding the role of lipophilicity and pharmacophore elements is paramount in
elucidating the inhibitory activity of pyrimidinedione-based compounds targeting BCAT1.
This study investigated a novel structural class of BCAT1 inhibitors, specifically
(trifluoromethyl)pyrimidinediones, identified through high-throughput screening. The research
emphasized the significance of lipophilicity, demonstrated by the increased BCAT1 inhibitory
action resulting from the substitution of lipophilic groups in the molecular structure.
Pharmacophore modeling revealed crucial elements, including hydrogen bond acceptors and
donors, influencing the BCAT1 inhibitory activity. The examination of potent inhibitors, such
as BAY-069, shaded light on the cellular activity and selectivity of these compounds. The study
contributes to a comprehensive understanding of the intricate relationship between
lipophilicity, pharmacophore features, and the effectiveness of pyrimidinedione-based BCAT1
inhibitors.
Keywords: Pharmacophore modeling, Pymol, BCAT1 Inhibitors, Pyrimidinedione,
Introduction:
The pursuit of effective cancer treatments has long been at the forefront of medical research,
prompting the exploration of innovative therapeutic avenues1. Among the promising targets for
cancer therapy are the branched-chain amino acid transaminase enzymes (BCAT1 and
BCAT2), which play a crucial role in the metabolic reprogramming of cancer cells. These
enzymes facilitate the catabolism of branched-chain amino acids (BCAAs), such as leucine,
isoleucine, and valine, leading to the generation of key metabolic intermediates necessary for
tumor growth and survival2.
In recent years, significant attention has been directed towards the development of BCAT1/2
inhibitors as potential anticancer agents. Among these, the pyrimidinedione-based inhibitors
have emerged as a particularly promising class of compounds with potent inhibitory activity
against BCAT1 and BCAT2 enzymes3.
The overexpression of BCAT1 and BCAT2 has been observed in various cancer types,
including breast, prostate, lung, and pancreatic cancers. Their upregulation has been associated
with aggressive tumor behavior, metastasis, and resistance to standard cancer therapies.
Consequently, targeting these enzymes represents a rational and attractive strategy to disrupt
cancer cell metabolism, impair tumor growth, and enhance the efficacy of existing treatments4.
Pyrimidinedione-based BCAT1/2 inhibitors offer several advantages over traditional
anticancer agents. Their unique chemical structure confers a high degree of specificity towards
BCAT1 and BCAT2 enzymes, minimizing off-target effects and reducing potential toxicity to
normal cells. Moreover, their distinct mode of action disrupts the intricate metabolic networks
within cancer cells, creating an environment detrimental to cancer cell survival5.
Pharmacokinetic profiling plays a pivotal role in drug development, guiding the understanding
of how a compound is absorbed, distributed, metabolized, and excreted in the body. This study
focuses on the pharmacokinetic profiling of innovative pyrimidinedione-based BCAT1
inhibitors, aiming to unravel essential aspects related to their bioavailability, tissue distribution,
and metabolic fate6. The investigation explores how these novel compounds interact within the

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biological system, with a specific emphasis on their potential implications for optimal drug
delivery and therapeutic efficacy. By elucidating the pharmacokinetic behavior of these
BCAT1 inhibitors, this research aims to provide valuable insights that can inform the design
of effective drug formulations, dosing regimens, and administration routes, ultimately
advancing their translational potential in the realm of cancer therapeutics.
Methodology:
1. Selection of Dataset: The consensus pharmacophore model was developed using a dataset
of 43 molecules7. The molecules were screened for their activity and selectivity for BCAT1
inhibitors. The activity values (IC50 expressed as nM) were used to find most active molecules.
The Table 1 contains top active molecules and least active molecules which are used for model
building.
Table 1. SMILES notations and activity values IC50 (nM) of four highly reactive molecules
and least active molecule used for alignment
S.N. SMILES IC50 (nM)

1 Cc1ccccc1Oc2cc(N3C(=O)NC(=CC3=O)C(F)(F)F)c4ccccc4c2Cl 31
COc1cc(C#N)c(Oc2ccccc2C)cc1N3C(=O)NC(=CC3=O)S(=O)(=O)c4c
2 cccc4 60

3 COc1cc(C#N)c(Oc2ccccc2C)cc1N3C(=O)NC(=CC3=O)C(F)(F)F 162

COc1cc(C#N)c(Oc2ccccc2C)cc1N3C(=O)NC(=CC3=O)C(F)(F)C(F)(F) 792
4 F

5 Cc1ccccc1Oc2ccc(F)c(c2)N3C(=O)NC(=CC3=O)C(F)(F)F 50000

2. Compound Representation, Geometry Optimization & File Format Conversion with

Open Babel::

 Chemical structures were drawn using Chem Draw ennsuring accurate depiction of bond
types, stereochemistry, and relevant functional groups.
 Optimized the 3D structures of the compounds using Chem Draw or a molecular
mechanics tool to refine bond angles and minimize steric clashes
 Identified key pharmacophoric features (e.g., hydrogen bond donors/acceptors,
hydrophobic regions) using Chem Draw.
 Annotated each feature with relevant distances and angles.
 Converted the optimized 3D structures to a common format (e.g., MOL) using Open
Babel for compatibility with PyMOL.8

3. Pharmacophore Alignment and Visualization with PyMOL:

 Loaded the converted structures into PyMOL for molecular visualization.
 Aligned the structures based on common pharmacophoric features using PyMOL's
alignment tools.
 Visualized and refined the pharmacophore model in PyMOL, considering spatial
arrangements.9

4. Validation and Refinement & Export Pharmacophore Features:

 Validated the pharmacophore model against known ligand-receptor interactions.
 Refined the model iteratively based on experimental data or additional computational analyses.

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 Exported the refined pharmacophore features from PyMOL for further analysis or virtual
screening.

(a) (b)
Figure 1. Consensus pharmacophore model (a)Most active Molecule (IC50=31nm) (b) Most
active molecule (IC50=5000nm) showing contours for different regions (Green: Lipophilic,
Blue: H-Bond donor and Red: H-Bond acceptor region)
Result and Discussions:
The current pharmacophore-oriented research reveals a strong association between the BCAT1
inhibitory activity of the compounds chosen for this investigation and five lipophilic, one H-
Bond donor, and four H-bond acceptor groups. However, as the pharmacophore modeling of
the other most active compounds listed in the above table shows, the mere presence of a given
number of lipophilic, donar, and acceptor regions does not immediately correlate with the level
of their activity against BCAT1 enzyme.
Increased lipophilicity of the molecules caused by the substitution of lipophilic groups for the
original molecule is thought to be the probable cause of the increased BCAT1 inhibitory action.
One nitrogen atom function as a donor and the other two oxygen atoms on the pyrimidinedione
ring as H-bond acceptors. According to published research, these medicinal compounds' ability
to bind BCAT1 inhibitors is directly impacted by regions that are more lipophilic.
Figure 1 examination demonstrates that the lipophilic character of the bicyclic and aromatic
rings of pyrimidinedione-based BCAT1 inhibitors is responsible for the BCAT1 inhibitory
activity of BAY-069 derivatives.

(a-1) (a-2)

Figure 2: Intramolecular distances (pm) of lipophilic region

Based on pharmacophore-oriented study, the selected compounds have five lipophilic, one H-
bond donor, and four H-bond acceptor groups, all of which are positively correlated with
BCAT1 inhibitory action. Yet, as previous highly active molecules have demonstrated, the

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fixed number of these regions alone does not directly correlate with the strength of BCAT1
inhibitory activity10.

Increased lipophilicity brought about by molecule replacement with lipophilic groups is

thought to be the cause of the increased BCAT1 inhibitory action. Increased lipophilic surfaces
affect the intramolecular distances of lipophilic regions, H-bond donors, and H-bond acceptors,
which in turn affects how certain BCAT1 amino acids interact (Figure 2).

Conclusions: The pharmacophore-oriented exploration of BCAT1 inhibitory activity reveals

a nuanced connection between molecular characteristics and compound efficacy. While the
presence of lipophilic, donor, and acceptor groups correlates positively with BCAT1 inhibition,
this relationship is not solely dictated by their fixed quantity. Pharmacophore modeling of
highly active compounds underscores the influence of additional factors on overall BCAT1
enzyme activity. Enhanced lipophilicity, achieved through substituting lipophilic groups,
significantly contributes to heightened BCAT1 inhibitory effects. This involves the
introduction of lipophilic moieties, increasing relevant surfaces, impacting intramolecular
distances, and influencing interactions with BCAT1 amino acids. The nitrogen and oxygen
atoms on the pyrimidinedione ring, serving as H-bond donor and acceptors, respectively,
emphasize the crucial role of these structural elements. In conclusion, this research underscores
the intricate interplay of molecular features in BCAT1 inhibitory activity, stressing the need
for a comprehensive understanding beyond the mere presence of specific pharmacophoric
elements. Future BCAT1-targeted drug design should consider quantitative aspects, spatial
arrangements, and interactions for potent inhibitor development.
References:
1
“Targeted Therapy Drug List by Cancer Type - NCI,” cgvArticle, December 20, 2023,
nciglobal,ncienterprise, https://www.cancer.gov/about-cancer/treatment/types/targeted-therapies/approved-
drug-list.
2
Kassidy Lee and Cynthia Blanton, “The Effect of Branched-Chain Amino Acid Supplementation on Cancer
Treatment,” Nutrition and Health 29, no. 4 (December 2023): 621–35,
https://doi.org/10.1177/02601060231153428.
3
Judith Günther et al., “BAY-069, a Novel (Trifluoromethyl)Pyrimidinedione-Based BCAT1/2 Inhibitor and
Chemical Probe,” Journal of Medicinal Chemistry 65, no. 21 (November 10, 2022): 14366–90,
https://doi.org/10.1021/acs.jmedchem.2c00441.
4
Mohammad Hassan Baig et al., “Enzyme Targeting Strategies for Prevention and Treatment of Cancer:
Implications for Cancer Therapy,” Seminars in Cancer Biology, Current Vision on Target Enzymes for
Cancer Therapy, 56 (June 1, 2019): 1–11, https://doi.org/10.1016/j.semcancer.2017.12.003.
5
Jennifer A. Borthwick et al., “Structurally Diverse Mitochondrial Branched Chain Aminotransferase
(BCATm) Leads with Varying Binding Modes Identified by Fragment Screening,” Journal of Medicinal
Chemistry 59, no. 6 (March 24, 2016): 2452–67, https://doi.org/10.1021/acs.jmedchem.5b01607.
6
Teresa Kaserer et al., “Pharmacophore Models and Pharmacophore-Based Virtual Screening: Concepts and
Applications Exemplified on Hydroxysteroid Dehydrogenases,” Molecules 20, no. 12 (December 19, 2015):
22799–832, https://doi.org/10.3390/molecules201219880.
7
Günther et al., “BAY-069, a Novel (Trifluoromethyl)Pyrimidinedione-Based BCAT1/2 Inhibitor and
Chemical Probe.”
8
“Open Babel - the Chemistry Toolbox — Open Babel Openbabel-3-1-1 Documentation,” accessed January
25, 2024, https://openbabel.org/.
9
“PyMOL | Pymol.Org,” accessed January 27, 2024, https://pymol.org/2/.
10
Günther et al., “BAY-069, a Novel (Trifluoromethyl)Pyrimidinedione-Based BCAT1/2 Inhibitor and
Chemical Probe.”

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53
Use of Formic Acid as Co Surrogates in Palladium Catalyzed
Carbonylation Reaction

Shoeb R. Khan*, Pratik E. P. Michael*, Mayur V. Khedkar*

Department Of Chemistry, Hislop College, Nagpur-440001 (M.S), India
E-mail: [email protected]; 9096312458

ABSTRACT:
Carbonylation is the process of utilizing carbon monoxide (CO) as a highly effective building
block in organic transformations. Nevertheless, managing this hazardous and environmentally
impactful CO gas poses a significant challenge for chemists. To address this concern, toxic
carbon monoxide gas has been substituted with alternative CO sources referred to as CO
surrogates. Formic acid and formaldehyde have proven to be highly applicable in organic
synthesis and are consequently utilized as non-gaseous CO surrogates. This article outlines
recent advancements in the exploration of formic acid as a CO surrogate in transition-metal
catalyzed carbonylation reactions.
Keywords: Carboxyboronate, Carbon building block, C-C coupling, Carbonylation

INTRODUCTION:
Transition-metal-catalyzed carbonylations represents an important class of reactions for
the effective preparation of range of carbonyl-containing compounds (Cornils 2009). The
Carbonylation reaction allows one step insertion of CO to a substrate therefore much efforts
has been made to explore the new carbonylative transformations (Wang 2010). The first
carbonylation reaction catalyzed by transition metal using gaseous carbon monoxide (CO) was
demonstrated Heck and co-workers in 1974 (Schoenberg 1974). This discovery set milestone
in use of gas phase reaction.
There is a major drawback with a carbonylation reaction is that in most cases, the highly
toxic, tasteless, odorless, and colorless gas carbon monoxide is required, and it is often
exploited in large excess and in many instances at higher pressure then the atmospheric (Cao,
2017). The laboratories also need special CO detectors and possibly high pressure reactors limit
the use of this CO gas (Khedkar, 2011). As a result of the physical properties of carbon
monoxide, its use in the fine chemical industry and academia is limited. Thus in context of
solving this problem, carbonylations without the use of CO gas become better alternative for
synthetic organic chemists.
In this respect, in 2004 Morimoto and Kakiuchi explored the CO surrogates for Pauson–
Khand reactions, carbonylation reactions by using aromatic halides, and sometime applied for
alkenes.(Morimoto, 2004) Since then, many research projects have addressed new strategies
and new carbonyl sources. (Park, 2010)
M. Larhed and co-workers reviewed the molybdenum hexacarbonyl as a solid surrogate
for the carbonylative insertion reactions (Odell, 2012). Beller et al. summarized different
carbonylation reactions using alkenes as a substrate with CO surrogates (Wu, 2014). Use of
formic acid derivatives like phenyl formate and N-formylsaccharin as CO surrogates was
demonstrated by Konishi and Manabe (Konishi 2014). Bhanage and co-workers focused on
alkynes, arenes and aryl halides as a substrate for carbonylation reaction catalyzed by

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transition-metal using CO surrogates (Gautam, 2015). Use of COgen (9-methylfluorene-9-

carbonyl chloride) and SilaCOgen (methyldiphenylsilacarboxylic acid) was explored by
Skrydstrup and co-workers (Friis, 2016). Unlike these CO surrogates, formic acid and there
salt also found to be an excellent source of CO. However, formic acid (HCOOH) was
successful applied carbonylation reactions. Hence present article address the recent updates in
use formic acid and there salt in CO free carbonylation reactions.
Use of Formic Acid in Palladium-Catalyzed Carbonylation reactions
Thus in search of new safer CO surrogates, for the first time Simonato discovered use
of formic acid as a CO source (Simonato, 2003). After this discovery first palladium catalysed
Carbonylation reaction using formic acid as alternative to CO was explored by Cacchi and co-
workers (Scheme 1) (Cacchi, 2003). They studied the hydroarylation reactions of alkynes with
aryl halides catalyzed by palladium by employing formate anions as reducing agents. They
found that in the presence of acetic anhydride, formate anion can act as CO surrogate. They
also projected a mechanism which comprising the formation of formic acetic anhydride from
formate anion and acetic anhydride. The reaction proceed with the thermal decomposition to
generate acetic acid and carbon monoxide, which underwent in-situ palladium-catalyzed
hydroxycarbonylation.

Scheme 1:- Palladium catalyzed carbonylation reaction using formic acid as a CO source.
Bessmernykh and Caille et al explored the combination of palladium(II) acetate with
1,1’-bis(diphenylphosphino)ferrocene(dppf) as a catalytic combination for
hydroxycarbonylation of aryl and vinyl bromides (Scheme 2) (Berger, 2006).

Scheme 2:- Palladium catalyzed hydroxycarbonylation using formic acid as a CO source.

Skrydstrup and co-workers discovered use potassium salt of formic acid (HCOOK) as
CO source for carbonylation reaction (Korsager, 2013). An acyl Pd(II)
complex was employed as a pre-catalyst with dtbpf as ligand. The reaction condition was tested
for different substates.

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The cross-coupling reaction involving CO is a powerful carbon-carbon bond formation

process which leads to synthesis of range of useful carbonyl compounds like ketones, Amides
or esters etc. Carbonylative Coupling of alkyne with aryl iodide i.e Sonogashira coupling using
formic acid as carbon monoxide surrogates has been reported by Wu and co-workers (Scheme
3). (Qi, 2015)

Scheme 3:- Palladium catalyzed Sonogashira coupling employing formic acid as CO surrogates.

Carbonylative Suzuki coupling employing reaction of aryl halides and arylboronic

acids in occurrence of formic acid was published by Qi et al. (Scheme 4). (Qi, 2015) Same
group further reported synthesis of ester using Palladium catalysed Alkoxycarbonylation under
Formic acid (Qi, 2016)

Scheme 4:- Palladium catalyzed Suzuki coupling with formic acid as CO surrogates.
Wu and co-workers explored Palladium-catalyzed reductive carbonylation of aryl
iodides with formic acid as the formyl source. This protocol gave direct access to wide range
of Aromatic aldehyde in single step (Scheme 5). (Wu 2016)

Scheme 5:- Reductive carbonylation of aryl iodides with formic acid by sing Palladium

Nozaki and co-workers developed Palladium-catalyzed hydroxycarbonylation of

arenes via aromatic C–H bond activation using formic acid as a source of carbonyl (Shibahara,
2004). During this Surprisingly, they found that, simultaneously hydroxylation and
carboxylation of biphenyl to give 4’-hydroxy-4-biphenylcarboxylic acid in the presence of
formic acid, whereas with carbon monoxide gas instead of formic acid, the reaction only give
hydroxycarbonylation product (Scheme 6).

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Scheme 6:- Hydroxycarbonylation of arenes via aromatic C-H bond activation using formic acid.
Shi and group developed hydrocarboxylation of olefins with HCOOH and HCOOPh
catalyzed by palladium. In this reaction HCOOPh was used in catalytic amount. Using same
reaction condition lactones and β-amino acid derivatives were also prepared (Scheme 7)
(Wang, 2014)

Scheme 7:- Hydroxycarbonylation of arenes via aromatic C-H bond activation using formic acid.
In 2015, Zhou and co-workers proposed hydrocarboxylation of alkynes and formic acid
catalyzed by palladium and xantphos. The reaction circumvents a novel method for the
synthesis of acrylic acids under mild carbonylation deprived of external CO gas. A catalytic
amount of acetic anhydride was utilized for release of carbon monoxide (Scheme 8) (Hou,
2015).

Scheme 8:- Hydrocarboxylation reaction of alkynes and formic acid with Palladium Xantphos.

Fu et al. further promoted Nickel catalysed hydrocarboxylation of various alkynes with

HCOOH, in which 1,2-bis(diphenylphosphino)-benzene (dppbz) or cis-1,2
bis(diphenylphosphino)ethylene (dppen) was used as ligand (Scheme 14) (Fu, 2016)
In 2018 selective palladium-catalyzed alkoxycarbonylation of all kinds of alkenes with
formic acid (HCOOH, FA) was described Sang et al (Sang. 2018). Different alkenes including
terminal, di-, tri-, and tetra-substituted olefins were transformed into linear esters with very
good yields and regioselectivity. Very recently hydrocarboxylation of terminal alkynes with
formic acid was reported by Liu and group (Liu, 2022) different α,β-unsaturated carboxylic
acids with excellent branched regioselectivity were prepared in one step (Scheme 9).

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Scheme 9:- Terminal alkynes with formic acid for Hydrocarboxylation raction
Xinxin Qi et al. first reported the synthesis of aurone using Formic acid as a CO
source.[Qi, 2016] In this strategy, in situ CO was generated using formic acid and acetic
anhydride which then employed in carbonylation of 2-iodophenol with terminal alkynes. They
also screen different Pd catalyst, solvents, and bases for optimization (Scheme 10).

Scheme 10:- Synthesis of aurone using Formic acid as a CO source.

CONCLUSION
This review highlights the recent advancements in carbonylation reactions, with a particular
focus on formic acid as a substitute for CO. The shift from toxic CO gas to a cost-effective,
readily accessible, and convenient CO source such as formic acid has garnered considerable
attention in recent times. The literature reveals diverse approaches, including cross-coupling
reactions of organic halides, hydroxycarbonylation, reductive carbonylation, and carbonyl
insertion into C-H bonds, showcasing the versatility and widespread application of formic acid
in these processes.

Reference
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Industrial Homogeneous Catalysis. Angew. Chem. Int. Ed. Engl, 33(21), 2144-2163.
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carboalkoxylation of aryl, benzyl, and vinylic halides. J. Org. Chem., 39, 3318–3326.
4. CAO, J.; ZHENG, Z.; XU, Z.; XU L. (2017,) Transition-Metal-Catalyzed Transfer
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5. KHEDKAR, M. V.; KHAN, S. R.; SAWANT, D. N.; BAGAL, D. B.; BHANAGE, B.M.
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carbonylative synthesis of N-substituted phthalimides. Adv. Synth. & Cat., (18), 3415–3422.
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carboalkoxylation of aryl, benzyl, and vinylic halides. J. Org. Chem. 39, 3318–3326.
7. MORIMOTO T., KAKIUCHI K., (2004) Evolution of Carbonylation Catalysis: No Need for
Carbon Monoxide, Angew. Chem. 43, 5580 – 5588.
8. PARK, J. H., CHO, Y., CHUNG, Y. K,. (2010) Rhodium-Catalyzed Pauson–Khand-Type
Reaction Using Alcohol as a Source of Carbon Monoxide, Angew. Chem. Int. Ed., 30, 5264-
5267.
9. SIMONATO, J.-P., (2003) New efficient catalytic system for hydroxycarbonylation without
CO gas, J. Mol. Catal. A 197, 61–64.

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10. ODELL, L., RUSSO, F., LARHED, M., (2012) Molybdenum Hexacarbonyl Mediated CO Gas-
Free Carbonylative Reactions, Synlett 23, 685-698.
11. WU, L., LIU, Q., JACKSTELL, R., BELLER, M., (2014) Carbonylations of Alkenes with CO
Surrogates Angew. Chem. Int. Ed. 53, 6310-6320.
12. KONISHI, H., MANABE, K., (2014) Formic Acid Derivatives as Practical Carbon Monoxide
Surrogates for Metal-Catalyzed Carbonylation Reactions, Synlett 25, 1971-1986.
13. GAUTAM, P., BHANAGE, B. M., (2015) Recent advances in the transition metal catalyzed
carbonylation of alkynes, arenes and aryl halides using CO surrogates Catal. Sci. Technol. 5,
4663-4702.
14. FRIIS, S. D., LINDHARDT, A. T., SKRYDSTRUP, T., (2016) The Development and
Application of Two-Chamber Reactors and Carbon Monoxide Precursors for Safe
Carbonylation Reactions Acc. Chem. Res. 49, 594–605.
15. CACCHI, S., FABRIZI, G., GOGGIAMANI, A., (2003) Palladium-Catalyzed
Hydroxycarbonylation of Aryl and Vinyl Halides or Triflates by Acetic Anhydride and Formate
Anions Org. Lett. 5, 4269–4272.
16. BERGER, P., BESSMERNYKH, A., CAILLE, J.-C., MIGNONAC, S., (2006) Palladium-
Catalyzed Hydroxycarbonylation of Aryl and Vinyl Bromides by Mixed Acetic Formic
Anhydride, Synthesis 3106-3110.
17. KORSAGER, S., TAANING, R. H., SKRYDSTRUP, T., (2013) Effective Palladium-
Catalyzed Hydroxycarbonylation of Aryl Halides with Substoichiometric Carbon Monoxide, J.
Am. Chem. Soc. 135, 2891- 2894.
18. QI, X., JIANG, L.-B., LI, C.-L., LI, R., WU, X.-F. (2015) Inside Cover: Palladium-Catalyzed
One-Pot Carbonylative Sonogashira Reaction Employing Formic acid as the CO Source, Chem.
Asian J. 10, 1802-1802.
19. QI, X., LIA, R., WU, X.-F. (2016) Selective palladium-catalyzed carbonylative synthesis of
aurones with formic acid as the CO source RSC Adv. 6, 62810-62813.
20. QI, X., LI, C.-L.,. JIANG, L.-B, ZHANG, W.-Q., WU, X.-F. (2016) Palladium-catalyzed
alkoxycarbonylation of aryl halides with phenols employing formic acid as the CO source,
Catal. Sci. Technol. 6
3099-3107.
21. QI, X., LI, C.-L., WU, X.-F. (2016) A Convenient Palladium-Catalyzed Reductive
Carbonylation of Aryl Iodides with Dual Role of Formic Acid, Chem. Eur. J. 22, 5835-5838.
22. SHIBAHARA, F., KINOSHITA, S., NOZAKI, K., (2004) Palladium(II)-Catalyzed Sequential
Hydroxylation−Carboxylation of Biphenyl Using Formic Acid as a Carbonyl Source, Org. Lett.
6, 2437–2439.
23. WANG, H., DONG, B., WANG, Y., LI, J., SHI, Y., (2014) A Palladium-Catalyzed
Regioselective Hydroesterification of Alkenylphenols to Lactones with Phenyl Formate as CO
Source; Org. Lett. 16 186-189.
24. HOU, J., XIE, J.-H., ZHOU, Q. L., (2015) Palladium-Catalyzed Hydrocarboxylation of
Alkynes with Formic Acid, Angew. Chem. Int. Ed. 54, 6302-6304.
25. FU, M. C., SHANG, R., CHENG, W. M., FU, Y., (2016) Nickel-Catalyzed Regio- and
Stereoselective Hydrocarboxylation of Alkynes with Formic Acid through Catalytic CO
Recycling, ACS Catal. 6, 2501–2505.
26. SANG, R., KUCMIERCZYK, P., DONG, K., FRANKE, R., NEUMANN, H., JACKSTELL,
R., BELLER M., (2018), Palladium-Catalyzed Selective Generation of CO from Formic Acid
for Carbonylation of Alkenes, J. Am. Chem. Soc. 140, 15, 5217–5223.
27. LIU, L., YAO, Y., CHEN, X., GUO, L., LU, Y., ZHAO, X., LIU. Y., (2022)
Hydrocarboxylation of alkynes with formic acid over multifunctional ligand modified Pd-
catalyst with co-catalytic effect. Journal of Catalysis , 405 , 322-332.

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54

In-Silico Prediction of Phytoconstituents From

Solanum Indicum for Antianxiety Activity
Shweta Dhole*, Parimal Katolkar, Pradeep Raghatate, Jagdish Baheti
Kamla Nehru College of Pharmacy, Butibori, Nagpur 441108 (MS), INDIA
*Email: [email protected]

ABSTRACT
Objective Anxiety is a common reaction to stress, and it may even be beneficial in certain
situations. It can alert us to potential dangers and help with planning and focus. Instead of the
usual sensations of apprehension or anxiety, there is excessive fear or anxiety when anxiety
disorders are present. Compounds found in medicinal plants have been the source of many
conventional medications. In-silico testing of Solaum indicum phytoconstituents for
antianxiety efficacy was a part of our investigation.
Methods Utilizing Discovery studio, molecular docking is done to assess the pattern of
interaction between the phytoconstituents from the Solanum indicum plant and the crystal
structure of the stress proteins (PDB ID: 5XVG). Later, SwissADME and pkCSM were used
to screen for toxicity as well as the pharmacokinetic profile.
Results The docked results suggest that Solafuranone (-8.1 kcal/mol) and Isofraxidin (-6.8
kcal/mol) for 5XVG macromolecule has best binding towards antianxiety activity as compared
to the standard (Gabapentin) for 5XVG is -5.1 kcal/mol. Furthermore, pharmacokinetics and
toxicity parameters were within acceptable limits according to ADMET studies.
Conclusion
Results from the binding potential of phytoconstituents aimed at antianxiety activity were
encouraging. It promotes the usage of Solanum indicum and offers crucial details on
pharmaceutical research and clinical care.
1. INTRODUCTION
In traditional Chinese medicine, the plant Solanum indicum is used to treat wounds and reduce
inflammation because it contains special dioscins, or indiosides. Our earlier research showed
that a number of synthetic indiosides derived from Solanum indicum has strong anticancer
properties, whereas other compounds have CNS depressing properties.1
Solanum indicum is a plant that is eaten as a vegetable in some parts of Africa, and folk
medicine experts claim that it may fend off cardiovascular conditions. The research on how it
can be utilised to treat hypertension was fascinating. The presence of chlorogenic acids may
only have served as a supportive role in the extract's strong therapeutic and preventive effects
against hypertension. Other ingredients could be responsible for the antihypertensive impact.
Future investigation into the extract as a potential therapy for hypertension is encouraged by
the results.2
However, there are few studies on the phytoconstituents of Solanum indicum for the antianxiety
activity. Thus, keeping the above information in view, the present investigation was designed
to identify the potential phytochemicals of Solanum indicum against 5XVG using a molecular
docking method.
2. MATERIALS AND METHODS
2.1. Platform for molecular docking
The computational docking study of all the phytoconstituents selected as ligands with
antianxiety activity as the target was performed using PyRx software.3
2.2. Protein preparation

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The macromolecule is 5XVG, in silico analysis of selected phytoconstituents was performed

on the 2.15 Å crystal structure of antianxiety macromolecule with inhibitor, (PDB ID: 5XVG,
having resolution Resolution: 2.10 Å, R-Value Free: 0.235, R-Value Work: 0.181, R-Value
Observed: 0.184), which was retrieved from the protein data bank (https://www.rcsb.org). 5XVG
is classified as crystal structure of PAK4 in complex with the inhibitor CZH226 all other
molecules, such as co-crystallized water molecules, unwanted chains, and nonstandard
residues, were deleted. Using Discovery studio.4
2.3. Mechanism of Action
5XVG: DI-1859 is a potent, selective and covalent DCN1 inhibitors. Mass spectrometric
analysis and co-crystal structures reveal that this compound employ a unique mechanism of
covalent bond formation with DCN1. The p21-activated kinases (PAKs) are serine/threonine
(Ser/Thr) protein kinases that have been identified as downstream signaling effectors of Rho-
family GTPases. PAK4 is the most studied group II PAK member, and it has a place at critical
nodal points in multiple signaling pathways that are associated with cell growth, cytoskeletal
dynamics, cell polarity, survival, and development. PAK4 is particularly highly expressed in
prostate, testis, lung, heart, brain, and liver.5
2.4. Ligand preparation
The three-dimensional (3D) structures of all constituents were retrieved using Avogadro
software from the PubChem database available on the NCBI website
(https://pubchem.ncbi.nlm.nih.gov/). However, the drawing of geometrical 2D structure was
performed using the ChemSketch program. The two-dimensional (2D) structures were
transformed into 3D models using the Avogadro software and the ligand structures were saved
in the PDB format. All the chemical structures are shown in Figure 1.

Solanidine Solasodine
OH H
H3C
H3C NH CH3
H3C H
H CH3
H3C
O
HH CH3
H
N H
HO

CH3

Solavetivone Solafuranone
CH2 H3C
CH3 H3C O
O
CH3
O
H3C CH3

CH3

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Scopoletin N-p-trans-Coumaroyltyramine

HO
H
N

O
OH

Isofraxidin Lauric acid

H3C O HO CH3

O
HO O O
O
H3C

𝛽-sitosterol Diosgenin
H
O
H3C

H3C H
H
H
O H

O H
H

Fig. 1. Chemical structures of all selected phytoconstituents in the molecular docking

studies
2.5. Standard Preparation
The standard is prepared steps such as, the 2D structure of standard drug was made using chem
sketch program, then the 2D structure was converted into 3D model using Avogadro Software,
it was saved in PDB format.
By using PyRx molecular docking of Gabapentin was done with 5XVG.
2.6. Molecular docking
Molecular docking evaluates the protein-ligand interactions and estimates the scoring function
based on the geometry to predict the binding affinity of the ligand molecule6,7. We
applied molecular docking studies to investigate the binding pattern of selected
phytoconstituents (Figure 1) and the standard drug, along with the crystal structure of
antianxiety activity macromolecule (PDB ID: 5XVG). The molecular docking study was
performed using PyRx software, Binding affinity was explored using the Vina wizard tool. The
final results were analysed and visualized using Discovery Studio 2020 Client 8, with bound
ligands as the standard. Visualization of protein ligand interaction reflects the number of
interactions and active residues responsible for significant binding at the active site of the target
enzyme.

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2.7. Absorption, distribution, metabolism, and excretion (ADME) and toxicity prediction
The selected phytoconstituents and standard drug were further checked for drug-likeness
properties according to Lipinski’s rule. During drug development, it is necessary to predict the
tolerability of phytochemicals before being ingested by humans and animal models.
The pharmacokinetic profile (ADME) and toxicity predictions of ligands were conducted using
SwissADME (http://www.swissadme.ch) and pkCSM (an online server database predicting
small-molecule pharmacokinetic properties using graph-based
signatures, (http://biosig.unimelb.edu.au/pkcsm/prediction). To analyse the toxicological
properties of ligands, Simplified Molecular Input Line Entry System (SMILES) notations or
PDB files were uploaded, followed by selecting the required models for generating numerous
information about structure-related effects9,10.
3. RESULT AND DISCUSSION
The present study aimed to explore the inhibitory potential of the phytoconstituents present
in Solanum indicum targeting antianxiety activity. In this study, we performed molecular
docking studies of all phytoconstituents found in Solanum indicum using AutoDock Vina,
followed by a study of interacting amino acid residues and their influence on the inhibitory
potentials of the active constituents. Selected phytoconstituents showing the best fit were
further evaluated for absorption, distribution, metabolism, excretion, and toxicological
(ADMET) properties using SwissADME and pkCSM servers.
3.1 Molecular docking
The docking scores and binding energies of all chemical constituents of Solanum
indicum targeting antianxiety activity (PDB ID: 5XVG) and binding interactions with amino
acid residues are presented in Table 1 respectively.

Table 1. Binding interaction of ligands from Solanum indicum targeting antianxiety

activity (PDB ID:5XVG)

Chemical Docking Score

Sr. No. PubChem ID
constituent 5XVG
1. Solanidine 65727 -8.0
2. Solasodine 5250 -8.3
3. Solavetivone 442399 -6.3
4. Solafuranone 11107208 -8.1
5. Scopoletin 5280460 -6.7
N-p-trans-
6. 5372945 -6.7
Coumaroyltyramine
7. Isofraxidin 5318565 -6.8
8. Lauric acid 3893 -4.2
9. 𝛽-sitosterol 222284 -7.4
10. Diosgenin 99474 -8.0
Standard Drug
11 Gabapentin 3446 -5.1

The binding affinities of phytoconstituents ranged from –8.3 to −4.2 kcal/mol. From the
docked results, it is evident that the compounds, Solasodine exhibit the most favourable binding
affinity (−8.3 kcal/mol) in complex with selected macromolecules (PDB ID: 5XVG) as
compared to other docked compounds i.e., Solafuranone (−8.1 kcal/mol), Solanidine (-8.0
kcal/mol), Diosgenin (-8.0 kcal/mol), , Beta sitosterol (-7.4 kcal/mol), Isofraxidin (−6.8
kcal/mol), Scopoletin (−6.7 kcal/mol), N-p-trans-Coumaroyltyramine (−6.7 kcal/mol),
Solavetivone (−6.3 kcal/mol), Lauric acid (−4.2 kcal/mol). Visual examination of the

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computationally docked optimal binding poses of phytoconstituents on selected

macromolecules (i.e., 5XVG) revealed the significant involvement of various types of
interactions, such as hydrogen bonding and hydrophobic interactions, including π–π stacking
and π–alkyl and alkyl interactions, in the stability of the binding of the phytoconstituents to
5XVG.
The binding affinity of the standard (Gabapentin) for 5XVG is -5.1 kcal/mol.
3.1.1. Solafuranone, 5XVG
An analysis of the interactions between the 5XVG protein complex and the Solafuranone ligand
was also carried out, which showed that the ligand molecule is oriented due to Pi-Sigma
interaction with LEU A:447 amino acid residue and Pi-Alkyl interaction with ALA A:348,
VAL A:335 amino acid residue were found.
3.1.2. Isofraxidin, 5XVG
An analysis of the interactions between the 5XVG protein complex and the Isofraxidin ligand
was also carried out, which showed that the ligand molecule is oriented due to Pi-Sigma
interaction with VAL A: 335, LEU A: 447 amino acid residue and Alkyl and Pi-Alkyl
interaction with ALA A: 348, MET A: 395, LYS A: 350 amino acid residue Conventional
Hydrogen bond with LEU A: 398, ASP A: 458 amino acid residue and Carbon hydrogen bond
with GLU A: 329 amino acid residue were found.
3.1.3. Gabapentin, 5XVG
The binding affinity of the standard (Gabapentin) for 5XVG is -5.1 kcal/mol. the interactions
between the 5XVG protein complex and the Gabapentin ligand was also carried out, which
showed that the ligand molecule is oriented due to conventional hydrogen bond with the ASP
A:444, SER A:443, GLU A:507 amino acid residue and attractive charge interaction with LYS
A:442 amino acid residue were found.
Table No. 2. Interactions with Amino acid residue
Bindin Main amino acid interactions
g Pi-alkyl, alkyl, Pi-S/Pi-Pi, Van der
Sr.
Energ T Waals
No Molecule H bond
y shaped/halogen/unfavour interaction
.
(kcal/ able donor-donor s
mol) interactions
PRO A: 578, ALA A:423, No
1 Solanidine -8.0 GLY A: 452
ALA A: 419 interactions
No PRO A: 479, TYR A: 480, No
2 Solasodine -8.3
interactions PHE A: 516 interactions
VAL A:
No
3 Solavetivone -6.3 454, ASN PRP A: 578, LEU A: 422
interactions
A: 377
No VAL A: 335, ALA A: 348, No
4 Solafuranone -8.1
interactions LEU A: 447 interactions
MET A:395, ALA A: 402,
No
5 Scopoletin -6.7 LEU A: 398 VAL A: 335, ALA A: 348,
interactions
LEU A: 447

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N-p-trans- GLU A:
ASP A: 458, SER A: 331, No
6 Coumaroyltyram -6.7 329, ASP
ILE A: 327 interactions
ine A:440
LEU A:
LEU A: 447, ALA A: 348,
398, ASP A: No
7 Isofraxidin -6.8 VAL A: 335, MET A: 395,
458, GLU interactions
LYS A:350
A: 329
PHE A: 304, ILE A: 369, VAL A: 368, ARG A:
8 Lauric acid -4.2
SER A: 300 PHE A: 364 360
VAL A: 335, LEU A: 447,
No No
9 𝛽-sitosterol -7.4 ALA A: 348, ILE A: 327,
interactions interactions
ILE A: 337
No No
10 Diosgenin -8.0 PRO A: 578
interactions interactions
ASP A: 444,
11 Gabapentin -5.1 SER A: 443, TRP A: 481 LYS A: 442
GLU A: 507

3.2. ADMET study

Pharmacokinetic profile (ADME) and toxicity predictions of the ligands are important attentive
parameters during the transformation of a molecule into a potent drug. In the present study,
these parameters were assessed using SwissADME and pkCSM. The absorption potential and
lipophilicity are characterized by the partition coefficient (Log P) and topological polar surface
area (TPSA), respectively. For better penetration of a drug molecule into a cell membrane, the
TPSA should be less than 140 Å. However, the value of Log P differs based on the drug target.
The ideal Log P value for various drugs are as follows: oral and intestinal absorption, 1.35 −
1.80; sublingual absorption, > 5; and central nervous system (CNS)11. The aqueous solubility
of ligands ideally ranges from − 6.5 to 0.5 12, while the blood brain barrier (BBB) value ranges
between − 3.0 and 1.2 13. In addition, non-substrate P-glycoprotein causes drug resistance 14.
In our study, all the selected ligands followed the TPSA parameter, P-glycoprotein non-
inhibition, thereby showing good intestinal absorption and an acceptable range of BBB
values. All the compounds showed aqueous solubility values within the range. Further, it was
predicted that the selected ligands do not show AMES toxicity, hepatotoxicity, and skin
sensitivity. In addition, it did not inhibit hERG-I (low risk of cardiac toxicity). Lipinski’s rule
violations, T. pyriformis toxicity, minnow toxicity, maximum tolerated dose, rat acute oral
toxicity, and chronic toxicity are depicted in table.

Table 3. ADME and toxicity predicted profile of ligands with superior docking score
Aqueou Human
Blood
MW HB Hb s intestina
ADMET TPSA Brain
Formula (g/mol Log P done accept solubili l
Properties (Ȧ2) Barri
) r or ty (Log absorpti
er
mol/L) on (%)
C27H43N 397.64 23.47
Solanidine 5.655 1 2 -4.927 92.975 0.695
O 7 Ȧ2

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C7H43N 41.49
Solasodine 413.64 5.2869 2 3 -3.809 92.324 0.035
O2 Ȧ2
17.07
Solavetivone C15H22O 218.33 3.9042 0 1 -4.615 95.873 0.635
Ȧ2
232.32 3.1876 26.30
Solafuranone C15H20O2 0 2 -3.551 95.523 0.206
3 4 Ȧ2
59.67
Scopoletin C10H8O4 192.17 1.5072 1 4 -2.504 95.277 -0.299
Ȧ2
N-p-trans-
C17H17N 283.32 69.56
Coumaroyltyram 2.4699 3 3 -3.165 90.031 -0.552
O3 7 Ȧ2
ine
222.19 68.90
Isofraxidin C11H10O5 1.5158 1 5 -2.458 95.588 -0.377
6 Ȧ2
200.32 37.30
Lauric acid C12H24O2 3.9919 1 1 -4.181 93.379 0.057
2 Ȧ2
414.71 20.23
𝛽-sitosterol C29H50O 8.0248 1 1 -6.773 94.464 0.781
8 Ȧ2
38.69
Diosgenin C27H42O3 414.63 5.7139 1 3 -5.713 96.565 0.2
Ȧ2
C9H17N 63.32
Gabapentin 171.24 1.3703 2 2 -2.778 93.257 -0.178
O2 Ȧ2

Table 3 Continued
Total Max
P-glyco- hERG
clearance Bioavailability AMES tolerated hERG I
ADMET protein II
[Log mL/ score toxicit dose inhibito
Properties substrat inhibito
(min.kg score y [Log mg/ r
e r
)] (kg.d)]
Solanidine YES 0.028 0.55 NO -0.882 NO YES
Solasodine YES 0.09 0.55 NO -0.375 NO YES
Solavetivone NO 1.225 0.55 NO 0.044 NO NO
Solafuranone NO 1.256 0.55 NO 0.526 NO NO
Scopoletin NO 0.73 0.55 NO 0.614 NO NO
N-p-trans-
YES 0.265 0.55 NO -0.213 NO YES
Coumaroyltyramine
Isofraxidin NO 0.713 0.55 NO 0.56 NO NO
Lauric acid NO 1.623 0.85 NO -0.34 NO NO
𝛽-sitosterol NO 0.628 0.55 NO -0.621 NO YES
Diosgenin NO 0.328 0.55 NO -0.559 NO YES
Gabapentin NO 0.525 0.55 NO 0.789 NO NO

Table 3 Continued
Oral rat Minno
T.Pyrifor
Acute oral chronic w
Skin mis Lipinski’s
ADMET rat toxicity. toxicity Hepatot toxicit
sensati toxicity rule
Properties LD50(mol/ (Log oxicity y (Log
on (Log Violation
kg) mg/kgbw/d mmol/
μg/L)
ay) L)
Solanidine 2.596 1.334 YES NO 0.378 -0.493 YES (1)
Solasodine 2.489 1.332 YES NO 0.311 0.381 YES (1)
Solavetivone 1.643 1.19 NO YES 1.453 0.874 YES (0)
Solafuranone 1.865 1.947 NO YES 2.151 0.557 YES (0)
Scopoletin 1.95 1.378 NO NO 0.516 1.614 YES (0)
N-p-trans-
Coumaroyltyra 2.17 1.271 NO NO 1.008 1.514 YES (0)
mine
Isofraxidin 2.326 1.825 NO NO 0.431 1.862 YES (0)

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Lauric acid 1.511 2.89 NO YES 0.954 -0.084 YES (0)

𝛽-sitosterol 2.552 0.855 NO NO 0.43 -1.802 YES (1)
Diosgenin 1.921 1.452 NO NO 0.399 0.247 YES (1)
Gabapentin 1.871 2.857 NO NO 0.265 1.698 YES (0)

3.3. Interaction of Standard Drug (Gabapentin) with 5XVG

Fig. 2. Docking scores and binding interaction of Gabapentin (PDB ID: 5XVG). The
ligand is shown in line and stick representation along with its 2D diagram and hydrogen
bond interaction.

3.4. Interactions of phytoconstituents with 5XVG

A.Solafuranone

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B.Isofraxidin

Fig. 3. Docking scores and binding interaction of phytoconstituents (PDB ID: 5XVG).
The ligand is shown in line and stick representation along with its 2D diagram and
hydrogen bond interaction.

3.5. Boiled Egg

Fig no. 4. Combined Boiled Egg Diagram

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Table no. 4. Name of molecules contained in Boiled Egg Diagram

MOLECULE NUMBER MOLECULE NAME
1 Solanidine
2 Solasodine
3 Solavetivone
4 Solafuranone
5 Scopoletin
6 N-p-trans-Coumaroyltyramine
7 Isofraxidin
8 Lauric acid
9 𝛽-sitosterol
10 Diosgenin
11 Gabapentin

BOILED means Brain Or IntestinaL EstimateD permeation predictive model. The boiled egg
diagram shows two regions white region and yellow region.
The white region is the physicochemical space of molecules with highest probability of being
absorbed by the gastrointestinal tract, and the yellow region (yolk) is the physicochemical
space of molecules with highest probability to permeate to the brain.
In addition, the points are coloured in blue if predicted as actively effluxed by P-gp (PGP+)
and in red if predicted as non-substrate of P-gp (PGP-).

4. CONCLUSION
In this study, we have carried out an in-silico screening of the phytoconstituents of Solanum
indicum plant. This study demonstrated the sixteen compounds from Solanum indicum plant,
(Solanidine, Solasodine, Solavetivone, Solafuranone, Scopoletin, N-p-trans-
Coumaroyltyramine, Isofraxidin, Lauric acid, 𝛽-sitosterol, Diosgenin). The selected
phytocompounds showed docking scores ranging from –8.3 to −4.2 kcal/mol in 5XVG. Among
all, Solafuranone and Isofraxidin gave the highest binding energy (−8.1 kcal/mol) and (−6.8
kcal/mol) in complex with 5XVG, whereas the reference compound, Gabapentin showed a
docking score with a binding energy of -5.1 kcal/mol. Furthermore, these ligands exhibited
good ADMET properties. To summarize, phytoconstituents present in Solanum indicum
possess strong effects against 5XVG and could be further evaluated for their antianxiety effect,
as well as for the development of alternative drugs with fewer side effects for the treatment of
stress.

REFRENCES
1. Ma P, Cao TT, Gu GF, Zhao X, Du YG, Zhang Y. Inducement effect of synthetic indiosides
from Solanum indicum L. on apoptosis of human hepatocarcinoma cell line Bel-7402 and
its mechanism. Ai Zheng= Aizheng= Chinese Journal of Cancer. 2006 Apr 1;25(4):438-42.

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2. Bahgat A, Abdel‐Aziz H, Raafat M, Mahdy A, El‐Khatib AS, Ismail A, Khayyal MT.

Solanum indicum ssp. distichum extract is effective against l‐NAME‐induced hypertension
in rats. Fundamental & clinical pharmacology. 2008 Dec;22(6):693-9.
3. Morris GM, Huey R, Lindstrom W, Sanner MF, Belew RK, Goodsell DS, Olson AJ.
AutoDock4 and AutoDockTools4: Automated docking with selective receptor flexibility.
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UCSF Chimera—a visualization system for exploratory research and analysis. Journal of
computational chemistry. 2004 Oct;25(13):1605-12.
5. Hao C, Zhao F, Song H, Guo J, Li X, Jiang X, Huan R, Song S, Zhang Q, Wang R, Wang
K. Structure-based design of 6-chloro-4-aminoquinazoline-2-carboxamide derivatives as
potent and selective p21-activated kinase 4 (PAK4) inhibitors. Journal of Medicinal
Chemistry. 2018 Jan 11;61(1):265-85.
6. Verdonk ML, Cole JC, Hartshorn MJ, Murray CW, Taylor RD. Improved protein–ligand
docking using GOLD. Proteins: Structure, Function, and Bioinformatics. 2003
Sep;52(4):609-23.
7. Leach AR, Shoichet BK, Peishoff CE. Prediction of protein− ligand interactions. Docking
and scoring: successes and gaps. Journal of medicinal chemistry. 2006 Oct 5;49(20):5851-
5.
8. SAMANT L, Javle V. Comparative docking analysis of rational drugs against COVID-19
main protease.
9. Arora S, Lohiya G, Moharir K, Shah S, Yende S. Identification of potential flavonoid
inhibitors of the SARS-CoV-2 main protease 6YNQ: a molecular docking study. Digital
Chinese Medicine. 2020 Dec 1;3(4):239-48.
10. Shah S, Chaple D, Arora S, Yende S, Moharir K, Lohiya G. Exploring the active
constituents of Oroxylum indicum in intervention of novel coronavirus (COVID-19) based
on molecular docking method. Network Modeling Analysis in Health Informatics and
Bioinformatics. 2021 Dec;10:1-2.
11. Kaloni D, Chakraborty D, Tiwari A, Biswas S. In silico studies on the phytochemical
components of Murraya koenigii targeting TNF-α in rheumatoid arthritis. Journal of Herbal
Medicine. 2020 Dec 1;24:100396.
12. Joshi T, Sharma P, Joshi T, Chandra S. In silico screening of anti-inflammatory compounds
from Lichen by targeting cyclooxygenase-2. Journal of Biomolecular Structure and
Dynamics. 2020 Aug 12;38(12):3544-62.
13. Nisha CM, Kumar A, Vimal A, Bai BM, Pal D, Kumar A. Docking and ADMET prediction
of few GSK-3 inhibitors divulges 6-bromoindirubin-3-oxime as a potential inhibitor.
Journal of Molecular Graphics and modelling. 2016 Apr 1;65:100-7.
14. Tsujimura S, Tanaka Y. Disease control by regulation of P-glycoprotein on lymphocytes
in patients with rheumatoid arthritis. World journal of experimental medicine. 2015 Nov
11;5(4):225.

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55
Computational Insight on the Argentophilic Interactions of Axially Chiral
Ag-NHC Complex of R-BINOL Scaffold via TD-DFT
Sonali Ramgopal Mahule*
ASAS, Amity University Mumbai, Panvel, India 410206
Abstract
A fascinating interaction between the closed shells of many organometallic or inorganic
compounds of d8, d2, and d10 systems is interestingly recognized as an important determinant
of solid-state structures and a potential source of useful material properties. This work presents
such kind of intramolecular metallophilic interactions in the X-ray diffraction structures of
bimetallic Ag-NHC complex 1e. The structural studies revealed that the geometry around the
metal centers has MM interactions. Time-dependent density functional theory (TD-DFT)
experiments were done for complex 1e to insight into the MM and metal-ligand interactions
in them. The results of computational and experimental studies of these complexes showed a
good agreement with each other. This attracting feature of closed-shell metallophilic
interactions is becoming extremely popular in a variety of materials including nanomaterials
for their applications that include rewritable phosphorescent paper for use as color-switchable
luminescent materials, phosphorescent organogels for reversible RGB-color switching,
luminescence chemosensors to optoelectronic ‘on-off’ devices, etc.
Introduction
The new chiral ligand framework has been derived from inexpensive and readily available,
enantiopure R-BINOL. This study uncovered some interesting cage-like structural features in
the molecular structure of the silver complex that have potential applications in important
domains like asymmetric catalysis and molecular recognition. Herein, we studied the
intramolecular MM interactions in the chiral bimetallic silver(I) NHC complex. In pursuing
this work we followed similar steps and concepts which were conspicuously studied and
reported in the literature for the silver(I) and gold(I) complexes of N/O-functionalized N-
heterocyclic ligand exhibiting closed-shell d10d10 intermolecular argentophilic and aurophilic
interactions.1,2 Whereas, this work differs from the reported one, and includes investigations
of the intramolecular argentophilic interactions in chiral NHC complexes of axially chiral
scaffolds containing amido functions. This property of metallophilic interaction in materials
is very much popular because of their various applications for eg. in chiral recognition, for the
development of chiral polymeric materials,3,4 in preparation of luminescent sensors5,6 etc. Thus,
the closed-shell metallophilic interactions by their subtle but significant influence have aroused
interest in recent years and have made an impact in frontline application-oriented research.
Results and Discussion
Time-dependent density functional theory (TD-DFT) studies
As the molecular structure of the complex [L(L'-NHC)2]Ag2 (L = 2,2-dioxo-binapthyl, L' =
phenyl-acetamido) (1e) is determined by X-ray diffraction studies (Figure 1) showed that these
are monomeric-bimetallic in nature and exhibit close MM contact of 3.3011(9) Å which is
shorter than twice the van der Waals radii of the corresponding silver (3.44 Å), there by
indicating the presence of closed shell d10d10 argentophilic interaction. Though numerous N-
heterocylic carbene complexes of silver are known in literature are examples of structurally
characterized discreet dimeric {(NHC)MX}2 (M =Ag, Au; X = halide)7,8 type. But the

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monomeric-bimetallic-bis-NHC complexes of type (Figure 1) exhibiting similar metallophilic

interaction84 are surprisingly not known to the best of our knowledge.

Figure 1. Monomeric chiral bis-MNHC type of complex showing MM interaction.

The geometry around the 1e [C1Ag1N1] = 164.09(10)o for 1e, [C1Ag1N3] = 167.5o is
consistent with the linear geometries often observed for two-coordinated d10 metals.9 Quite
interestingly, a direct consequence of presence and absence of MM (M = Ag) interaction
occurring through the central metal atom (Ag1) in the monomeric-bimetallic complex is
evidenced by the slightly bent angle [C1Ag1N1] = 164.09(10)o at silver(I) in [L(L'-
NHC)2]Ag2 (L = 2,2-dioxo-binapthyl, L' = phenyl-acetamido)10 (1e) and these results suggest
the presence of argentophilic interaction in complex 1e. The effect is very weak as these
frameworks are laid quite away from the metal binding sites of the ligands.
Table 1. Singlet excitation energies computed for complexes L(L'-NHC)2]Ag2 (L = 2,2-dioxo-
binapthyl, L' = phenyl-acetamido) (1e) and L(L'-NHC)2]Ag2 (L = 2,2-dioxo-binapthyl, L' =
phenyl-acetamido) 3e at B3LYP/SDD, 6-31G* level of theory with an output are shown.

Excitation Oscillator Type of

Transition
Complex Wavelength ψo-ψv
strength coefficient transition
(nm)

347.15 0.0905 HOMO→LUMO+1 0.6941 MLCT (π→π*)

339.43 0.0061 HOMO→LUMO+2 0.7002 MLCT (π→π*)

329.09 0.0064 HOMO→LUMO+3 0.6986 MLCT (π→π*)

Furthermore, to gain a better understanding about the interactions in the silver 1e complex, and
to obtain a deeper insight on the nature of NHC-metal interactions in these complexes, detailed
density functional theory studies were carried using coordinates of the X-ray diffraction
structure as an input data. In particular, the MM bond order (Wiberg Indices) was computed
using B3LYP/SDD, 6-31G(d) level of theory with the objective of estimating the strength of
the closed-shell metallophilic interaction present in the complex. The computed bond order
(Wiberg Indices) for the MM interaction in the silver complex 1e (0.1247) complex were
approximately in the order of about a fourth of a single covalent thus supporting the evidence
obtained from the X-ray diffraction analysis. The vertical transitions of the singlet excitation

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energies obtained using TD-DFT calculations at the B3LYP/SDD, 6-31G(d) level of theory
gives three transitions peaks for complex 1e (347 nm, 339 nm, 329 nm) which coincides with
the experimental results of UV-visible spectrum for 1e and showed absorption at 340 nm.
Table 2. Selected molecular orbitals of the complex L(L'-NHC)2]Ag2 (L = 2,2-dioxo-binapthyl,
L' = phenyl-acetamido) (1e) are shown.

HOMO Orbital # 203 HOMO Orbital # 204

HOMO Orbital # 205 LUMO Orbital # 206

TD-DFT study was performed on the geometry optimized computed structure, there can be a
change in positions of transitions wavelengths, when the same experiments will be performed
using X-ray crystallographic coordinates of the same complex. The number of transitions can
also be increase by increasing the number of microstates in the experiment for the detailed
study. These transitions obtained by the TD spectrum in 1e complex has been assigned to the
HOMO→LUMO π→π* MLCT transitions based on the comparison of intensities of the
observed transitions and the comparable computed transitions along with their oscillatory

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strength values10 (Table 1). Different molecular orbitals which are associated with transitions
of HOMO→LUMO for Ag-NHC complex 1e (Table 2) are both ligand and metal in character.

Conclusions
Silver(I) complex 1e showed mild range of metallophillic interactions in its structure. DFT
studies performed on the complex 1e showed good agreement with that of the experimental
studies of the complex. These studies further followed by TD-DFT studies which enabled to
determine the metalmetal and metal-ligand interactions, that also show different types of
HOMO→LUMO transitions.

References
1. Ray, L.; Shaikh, M. M.; Ghosh, P. Inorg. Chem. 2008, 47, 230.
2. Samantaray, M. K.; Pang, K.; Shaikh, M. M.; Ghosh, P. Inorg. Chem. 2008, 47, 4153.
3. Mercs, L.; Albrecht, M. Chem. Soc. Rev. 2010, 39, 1903.
4. Sluch, I. M.; Miranda, A. J.; Elbjeirami, O.; Omary, M. A.; Slaughter, L. M. Inorg. Chem.
2012, 51, 10728.
5. Cui, Y.; Yue, Y.; Qian, G.; Chen, B. Chem. Rev. 2012, 112, 1126.
6. Brandys, M.-C.; Jennings, M. C.; Puddephatt, R. J. J. Chem. Soc., Dalton Trans. 2000, 4601.
7. de Frémont, P.; Scott, N. M.; Stevens, E. D.; Ramnial, T.; Lightbody, O. C.; Macdonald, C.
L. B.; Clyburne, J. A. C.; Abernethy, C. D.; Nolan, S. P. Organometallics 2005, 24, 6301.
8. Wang, H. M. J.; Chen, C. Y. L.; Lin, I. J. B. Organometallics 1999, 18, 1216.
9. Cotton, F. A.; Wilkinson, G.; Murillo, C. A.; Bochmann, M. Advanced Inorganic Chemistry,
6th ed. 1999, p 1085.
10. Mahule, S.R.; Gangwar,M. K.; Vishnoi P. ChemistrySelect, 2018, 3 (14), 4023.

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56

Agricultural Influence and application of Mn(III), Fe(III) and VO(IV) Schiff

base metal Complexes

S. R. Kelode
Department of Chemistry, Arts, Commerce and Science College, Maregaon
e-mail: [email protected]

Abstract:
The newly synthesized complexes of Mn(III), Fe(III) and VO(IV) with Schiff base derived
from 2-hydroxy-5-methyl acetophenone and 4-(p-hydroxyphenyl)-2-aminothiazole have been
synthesized and characterized on the basis of elemental analysis, Infrared, 1HNMR, molar
conductance and magnetic susceptibilities. The Schiff base acts as a monobasic bidentate
ligand commonly coordinates through the oxygen atom of phenolic -OH group and the nitrogen
atom of azomethine group, which is confirmed by IR spectral data. Agricultural influence and
application of Schiff base metal complexes.
Keywords: Schiff base, Magnetic susceptibility and Agricultural application

Introduction:
Schiff bases are considered to be the most versatile ligands as they form complexes with the
metal atoms. Recent Advances in year 2023 the chiral Schiff Base Compounds 1 There is
synthesis, characterization and biological activities of new Schiff Base Compound and its
lanthanide complexe2. Antifungal Activity of Some Mixed Ligand Complexes Incorporating
Schiff Bases3 The aim of present investigation is to synthesize Mn(III), Fe(III) and VO(IV)
metal complexes of Schiff base derived by the condensation of 2-hydroxy-5-methyl
acetophenone and 4-(p-hydroxyphenyl)-2- aminothiazole and to check their potency towards
various agricultural influence and application of Schiff base metal complexes.

Experimental:
All the chemicals were of A.R. grade and used as received. 2-hydroxy-5-methyl acetophenone
(HMA) and 4-(p-hydroxyphenyl)-2 amino thiazole was prepared by known methods.4 -7 The
solvents were purified by standard methods.8
Synthesis of 4-(p hydroxyphenyl)-2 amino thiazole; A mixture of p-hydroxy acetophenone
(12g), thiourea (15.2g), iodine (25g), and 60ml distilled water was taken in a 250ml R.B. flask.
The mixture was refluxed for about 4h and poured into crushed ice to get the solid which is
basify with sodium hydroxide. The product was filtered and crystallized from 70% ethanol,
after several minutes the golden coloured product of 4-(p-hydroxyphenyl)-2 amino thiazole
was separated out.5 - 7 Yield: 10g (83.33%), m.p.: 2160C
COCH3 S
Refluxed N
NH2 C NH2 I2
OH 4H NH2
OH S
4-hydroxy acetophenone
4-(p -hydroxyphenyl)-2 amino thiazole

Synthesis of 2-hydroxy-5-methyl acetophenone 4-(p-hydroxyphenyl)-2 imino thiazole

[HMAT]: A solution of 4-(p-hydroxyphenyl)-2 imino thiazole (0.02M) in 25ml of ethanol was
added to an ethanolic solution(25ml) of 2-hydroxy-5-methyl acetophenone (0.02M) and the
reaction mixture was refluxed on a water bath for 4h. After cooling a pale yellow coloured
crystalline solid was separated out. It was filtered and washed with ethanol, crystallized from

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DMF and dried under reduced pressure at ambient temperature. The purity of ligand was
checked by elemental analysis and m.p. It was also characterized by IR and 1H NMR spectral
studies.
Yield: 60%; m.p. 3100C

OH OH
N Refluxed
N
4h
H3C COCH3 OH S NH2 H3C C N S OH
CH3 HMAT
2-hydroxy-5-methyl acetophenone 4-(p hydroxyphenyl)-2 amino thiazole

Table1. Analytical data of the Ligands.

Sr. Ligand Molecular Formula Colour and
Elemental Analysis
No. Formula Weight C% H% nature
N%
found Found Found
(Cal.) (Cal.) (Cal.)
1. HMAT C18H16N2O2S 324.16 Yellow 66.64 04.69 08.60
Crystalline (66.75) (04.93) (08.63)
Preparation of complexes: All the metal complexes were prepared in a similar way by
following method. To a hot solution of ligand HMAT (0.02M) in 25ml of ethanol a suspension
of respective metal salts was added drop wise with constant stirring. The reaction mixture was
refluxed on a water bath for 3-6 h. The precipitated complexes were filtered, washed with
ethanol followed by ether and dried over fused calcium chloride. Yield: 45-55%
The complexes are soluble in DMSO and DMF but insoluble in water and common organic
solvents. The metal chloride content of complexes were analyzed by standard methods.9
The 1H NMR spectra of ligand was recorded and obtained from RSIC Chandigarh. IR spectra
of the compounds were recorded on Perkin Elmer 842 spectrophotometer in the region 400-
4000cm-1, Carbon, Hydrogen and Nitrogen analysis were carried out at RSIC, Punjab
University, Chandigarh. The molar conductance of the complexes at 10-3 M dilution in DMF
were determined using equiptronic digital conductivity meter EQ-660 with a cell constant 1.00
cm-1 at room temperature. The magnetic moment measurement were made on a Gouy balance
at room temperature using [HgCo(SCN)4] as the calibrant.

Table 2. Analytical data and molar conductance of the compounds.

Compounds Colour Mol. Analysis % µeff M
wt. Found (Ω-1
(calc.) B.M. cm2
M C H N Cl mol-1)

[MnL2 (OAc)] 2H2O Brown 6.73 57.10 4.52 7.00 4.3 22.2
796.1 (6.89) (57.27) (4.64) (7.03) _

[FeL2 (H2O)Cl] H2O Brown 7.14 55.74 4.31 7.14 4.49 5.3 24.8
773.6 (7.22) (55.84) (4.39) (7.23) (4.58)

[VOL2] 7.11 60.45 4.09 7.76 1.55 10.8

Green 713.2 (7.15) (60.57) (4.20) (7.85) _

Result and Discussion:

The ligand and its complexes have been characterized on the basis of 1H NMR, IR spectral
data, elemental analysis, molar conductance and magnetic susseptibility measurements data.
All these values and analytical data is consistent with proposed molecular formula of ligand.
All the compounds are coloured solid and stable in air. They are insoluble in water but soluble

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in coordinating solvents like DMF and DMSO. The molar conductance values in DMF(10-3 M)
solution at room temperature (Table 2 ) shows all the complexes are non electrolytes.
The 1H NMR spectra of the ligand shows signals at δ 12.03, (1H, s phenolic OH ), δ 9.00 (1H,
s, phenolic OH ), δ 6.83, 6.82, 6.81 and 6.80 (4H, m, phenyl) δ 7.58, 7.53, and 7.52 (3H, s
Phenyl), 6.49 (1H s thiophene), and 2.92(3H, s, methyl) 2.58 (3H, s, methyl)10-13. The IR
spectral data of the ligand and its metal complexes are given in Table 3. The IR spectrum of
the free ligand exhibits a broad band at 3119 cm-1 which is ascribed to (O-H) vibration
lowered due to intramolecular O-H……N hydrogen bonding. The absence of C=O band at
around 1700 – 1750 cm-1 indicates the Schiff base formation.14-17 The other bands at 1620, 1513
and 1121 are assignable to (C=N), (C-O)(phenolic) and (C-S). In the spectra of all the
complexes the (C=N) band is lowered by 25-40 cm-1 indicating the involvement of
azomethine nitrogen in coordination. Also, the band due to (O-H) of the ligand disappeared
indicating the coordination of phenolic oxygen to metal ion via deprotonation. This is also
supported by the shift in (C-O) (phenolic) mode. IR spectrum of Mn(III) complex display two
bands near 1616, 1446 cm-1 assigned to asy(OCO) and sym(OCO) respectively, indicating
monodentate nature of the coordinated acetate group. The strong IR bands around 960, 1156
and 910 in the spectrum of VO(III), are assigned to (V=O) (Zr-OH) (O=U=O) vibrations.

Table 3. IR spectra of ligand and metal complexes:

v(O-H) hydrogen v(C=N) v(C-O) v(M-O) v(M-N)
Compound bonded Imine phenolic
HMAT 3119 1620 1513 - -

[MnL2 (OAc)] 2H2O _ 1560 1450 500 425

[FeL2 (H2O)Cl] H2O _ 1590 1502 520 455

[VOL2] _ 1594 1508 515 450

Agricultural Influence and application of metal complexes: A ploughshare is an cobalt Copper

and nickel metal complexes used in agriculture to increase the production of crops. They are
used prior to the seedling stage to loosen up the soil. Essential observed that cobalt (Co), copper
(Cu), iron (Fe), manganese (Mn), molybdenum (Mo), nickel (Ni), and zinc (Zn) complexes
with schiff base plays a beneficial role in plant growth and development. Synthetic Action on
Insecticides: Schiff base ligand was derived from the condensation of thiazole and 2-hydroxy-
5-chloro-3-nitro acetophenone. Their metal complexes show high activity against insects18. 2-
amino-4-phenylthiazole acts as intermediate in synthesis of photostable pyrthriod
insecticides19. Flourination on aldehyde part of Schiff base increases insecto acracicidal
activity20. Schiff bases and their metal complexes with Mn(III), Fe(III) and VO(IV) show
insecticidal activities against bollworm 21. Most metals complexes cause environmental and
atmospheric pollution, and may be lethal to humans. Heavy metals can become strongly toxic
by mixing with different environmental elements, such as water, soil, and air, and humans and
other living organisms can be exposed to them through the food chain. Arsenic, cadmium,
chromium, copper, nickel, lead and mercury are the most common metals which can pollute
the environment. They are not all equally important but all play a role in plant growth. The
heavy metals Mn(III), Fe(III) and VO(IV) are micronutrients or trace elements for plants. They
are essential for growth and stress resistance as well as for biosynthesis and function of
different biomolecules such as carbohydrates, chlorophyll, nucleic acids, growth chemicals,
and secondary metabolites. Nitrogen (N) containing Schiff base metal complexes is made

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available to the plant from the air and soil. But most of the needed elements that are nutrients
for plants come from the soil.

Conclusions: The Schiff bases ligand and their metal complexes shows more activity towards
the agricultural Influence and applicable for plant growth.

References:
1. T. China, N. Daisuke and A. Takashiro, Molecules, 28, 7990 (2023).
2. Y. Abu, A.A Abduh, M.S. Saghir and S.A.M Al-Gabri, J Pharma 15, 454, (2022).
3. M. Miloud, M. El-ajaily, Al-noor and N.S. Al-barki. J Bact Mycol. 7(1),1122 (2020).
4. A. S. Aswar, P.Bahad, A.Pardhi and N.Bhave, J. Poym. Mater. 5, 232 (1988).
5. S.Pattan, M.Ali, J.Pattan, S.Purohit, V.Reddy and B.Nataraj, Indian J.Chem., 45B, 1929. (2006).
6. D.Khrustalev, A.Suleimenova .and S.Fazylov, Russian J. App. chem., 81(5), 900 (2008).
7. H.Maradiya, and V.Patel, J. Fibers and poly., 3(1), 43 (2002).
8. B.Furniss, A.Hannaford, P.Smith & A.Tatchell, Vogel’s practical organic chemistry 5 thEd.
(Logman Scientific Technical, John Wiley and Sons), (1989).
9. Vogel AI, “A Text book of quantitative inorganic chemistry” 3 thEd., (ELBS,London,1961).
10. S.Sadigova, A.Magerramov and M.Allakhverdiev, Russian J. Org. chem., 81(5), 900 (2008).
11. E.Campbell and S.Nguyen, J. Tetrahedron, 42, 1221 (2001).
12. P.Pietikainen and A.Haikarainen J. Mole. Catalysis., 180, 59 (2002).
13. M.Kidwai, P.Poddar and k.Singhal, Indian J. Chem., 48B, 59 (2009).
14. S.Sonwane, S.Srivastava and S.Srivastava, Indian J. Chem., 47B, 633 (2008).
15. K.Patel and A.Mehata, E. J. Chem., 3(13), 267 (2006).
16. R.Maurya, D.Antony, S.Gopinathan, V.Puranic, S.Tavale &C.Gopinathan, Bull.Chem. Soc.,
68,2847 (1995).
17. D.Boghaei and S.Mohebi J. Tetrahedron, 58, 5357 (2002).
18. D.A. Laidler, D.J Milner, J. Organomet Chem., 1984 270, 121-129.
19. N. S. Kozlow, G.P. Korotyshova and N.G. Rozhkova, E.I.Andreeva. Chem Abstr. 106, 155955,
(1987).
20. L. Zhu, N Chen , .F. Li H, Song, X. Zhu, Chem Abstr,, 2004,141, 374026, (2004).
21. S. Huneck, K. Schreiber, H.D. Grimmecke , J Plant Growth Regul, 3,75,(1984).

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57
Recent Applications of Deep Eutectic Solvent (DES), a Greener Media in
Organic Synthesis
Suchita B. Wankhede*
Department of Chemistry, Amolakchand Mahavidyalaya, Yavatmal 445001 (Maharashtra) India
*Email – [email protected]
Mobile Number - 9421770305

Abstract: A modern era of research mainly focused on study of use of solvent in organic
transformation, with its advantageous as environmental benign, cost effectiveness and in a
simple and sophisticated practical term. In the search of safe, non-toxic, cheap, biorenewable
and biodegradable solvents, deep eutectic solvents (DESs) get the prominent position. DES
acts as a valuable alternative which overcome the limitations of use of traditional volatile
organic compound (VOCs) solvents. This review paper aims to inspire scientists, work in
organic field to study the potential of DES. From understanding of the different aspects like
effect of temperature, concentration, addition of water etc. would provide knowledge for
deciding the scheme of organic synthesis. Review paper includes study of various organic
reactions from literature using DESs as catalyst and solvent media in a tabular form. It also
includes recycling and reuse of DESs in organic reactions. Convenient reaction set-up, simple
separation and purification built impact of DESs in laboratory and industry in near future.
Keywords: Deep eutectic solvent, catalyst and solvent media, green/ecofriendly solvent,
organic transformation, recycle and reuse.
1. Introduction
“Protecting our common environment" is the key objective given in Millenium
Declaration adopted by United Nations in 2000. From the last decades when we throw a light
on our changing lifestyle which becomes more demanding in different fields like drugs, plastic,
polymer etc. These demand causes damage and exploitation of natural resources like water,
petroleum, air, minerals etc and the question was arises ‘Why is chemical manufacture
becoming unsustainable?’[1]. Now it becomes needed to make balance between increasing
demand in production of chemical compounds with safe and clean environment. Amongst three
states of matter, the most convenient reaction media is solvent (liquid state). Most common
volatile organic compound (VOCs) solvents, due to their high cost, high toxicity, high
inflammability, non-biodegradability and causing negative impact on environment[2], there is
need to develop a new concept of “ideal green solvent”. In search of ideal solvents, water,
supercritical carbon dioxide (SCD) [3] supercritical water (SCW)[4], and ionic solvents [5] are
discovered but due to their disadvantages under different reaction conditions make them
unsuitable solvent as reaction media for industrial purpose. Abbott in 2003 [6] first introduced
a new generation of safe, nontoxic, cheap, biorenewable and biodegradable, sustainable
designer solvent [7] termed as “Deep Eutectic Solvent” (DES). They exhibit similar properties
like ILs as they work on similar molecular base but still more superior than Ionic liquids. As a
designer and environmentally benign solvent, DES has numerous applications in various fields.
There are many reviews to study organic reactions based on greener approach using DESs as
reaction media. That encourages us to study organic reactions using greener solvent DES.
This review article includes general introduction of DESs, classification and types of
DESs, applications in various fields and study of various organic reactions using DESs as

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catalyst and solvent media. This review also discussed optimized reactions with recycling and
reuse of DESs in organic reactions from literature published in between the session 2020-2023.
2. Deep Eutectic Solvents (DESs)
Deep Eutectic Solvents (DESs), also known as Deep Eutectic Ionic Liquids (DEILs)[8],
Low Melting Mixtures (LMMs) or Low Transition Temperature Mixtures (LTTMs)[9] in the
literature. The first reported DES was a mixture of choline chloride and urea (1:2), with a
melting temperature of 12 ºC, much lower than those of the starting materials, 302 ºC and 133
ºC, respectively [10]. DES is generally defined as a binary mixture of an ionic components,
one is a hydrogen bond acceptor (HBA) like chloride ion and the other a hydrogen bond donor
(HBD), such as an alcohol, amide or carboxylic acid. Simply DES is a neoteric solvent [11]
which are able to form a new eutectic phase (which is a liquid below 100 °C) through hydrogen-
bond-promoted self-association and no covalent compounds are formed between them, hence
the term ‘Deep’ is used [12,13].
3. Types and Classification of DESs
DESs are broadly classified as Hydrophilic and Hydrophobic DESs. Main difference
between them is their melting point depression. Hydrophilic DESs have deep depression in
freezing point due to extensive hydrogen bond interaction eg. choline chloride salt with small
alkyl chains, acids, amines etc. While hydrophobic DESs have a large and small depression in
freezing point eg. Long hydrocarbon alkyl chain containing salts and acid like Decanoic acid,
Dodecanoate sodium salts etc.
DESs are also classified into four types (table 1) from numerous possible combinations
of HBD and HBA components. Marcus [14] reported data regarding their molecular mass and
their eutectic melting points. Nonionic deep eutectic solvents are the other class of DESs,
introduced by Francisco et al [15,16] originated from Type –III DES and are characterized by
low iconicity, having low electrical conductivity.

Table 1. Types of eutectic mixtures

Types General formula Terms Example
DES I Cat+X-+zMClx M=Zn, In, Sn, Al, Fe ChCl+ZnCl2
DES II Cat+X-+zMClx M=Cr, Ni, Cu, Fe, Co ChCl+CoCl2.6H2O
DES III Cat+X-+zRZ Z=OH, COOH, CONH2 ChCl+urea
DES IV MClx+zRZ M=Zn, Al and Z=OH, CONH2 ZnCl2+urea

4. Various Organic reactions in four types of deep eutectic solvents

4.1 DES Type I catalysed organic reactions
DES type –I is the combination of quaternary ammonium salts with metal chlorides, eg.
quaternary ammonium salts forms eutectic mixtures with zinc and tin chloride to result in
freezing points between 13 and 92 °C [17] respectively.

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Table 2. DES type I reactions

S. DES used General reaction and reaction condition Scope of No. of Refer
N. reaction recycle ences
(%yield) of DES

1. [CholineCl] 45-95% 3 [18]

[ZnCl2]3
2. [ChCl][ZnCl2] 30-81% 5 [19]

3. [ChCl][ZnCl2] 75.1- 3 [20]

92.6%

4. [ChCl][ZnCl2]2 63-98% - [21]

4.2 DES Type II catalysed organic reactions

DES type II involves organic salts that are easy to handle and overcomes moisture absorption
problem in DES type I. Eg. Choline chloride forms a deep eutectic solvent with calcium
chloride hexahydrate at several molar ratios, at a 1:2 molar ratio, melting point 2.70 ºC [22].

Table 3. DES type II reactions

S. DES used General reaction and reaction condition Scope of No. of Refer
N. reaction recycle ences
(%yield) of DES

1. ChCl.AlCl3 29-86% 10 [23]

.6H2O

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4.3 DES Type III catalysed organic reactions

Type III DESs are very attractive and most investigated class of DES as they are composed of
biodegradable and non-toxic HBD’s like urea, organic carboxylic acids, polyols. Eg.
Tetrabutylammonium chloride forms deep eutectic solvents with ethylene glycol (1:3 mole
ratio), glycerol (1:4 mole ratio), and triethylene glycol (1:3mole ratio) with eutectic freezing
point −31 °C, −13 °C and −43 °C, respectively [24].
Table 4. DES type III reactions

S. DES used General reaction and reaction condition Scope of No. of Refer
N. reaction recycle ences
(%yield) of DES

1. CholineCl:Urea 84-92% 4 [25]

2. Ethyltriphenylp 60-95% 5 [26]

hosphoniumbro
mideTHFtetraca
rboxylic acid.
3. ChCl:Urea (1:2) Stir.(20- 4 [27]
76)MW(1
2-49)
US(17-
64%)

4. [ChCl][Gabape 86-95% 5 [28]

ctine]

5. ChCl:Urea (1:2) 84-97% 4 [29]

6. ChCl:Urea (1:2) 84-90% 5 [30]

7. Taurine/choline 87-95% 4 [31]

chloride

88-94%

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4.4 DES Type IV catalysed organic reactions

Organic salts used in DES type I, II and III as HBA are now replaced by metal halides in
DES type IV. Urea:ZnCl2 (3.5:1mole ratio) forms a DES with a melting point of 9 °C [32].
Table 5. DES type IV reactions
S. DES used General reaction and reaction condition Scope of No. of Refer
N. reaction recycle ences
(%yield) of DES

1. CeCl3·7H2O/ Diketone( 5 [33]

urea (1:5) 80-
96%)Xant
henedione
(88-97%)

2. ZrOCl2·8H2O/ 91-99% 3 [34]

Urea (1:5)
3. K2CO3/Glycerol 85-95% - [35]
(1:4)

4. ZrOCl2·8H2O: 88-98% 5 [36]

Urea (1:5)

5. Conclusion
DESs have aroused increasing scientific attention in the field of greener chemistry due
to their easy availability and cost effectiveness. Study of organic reactions under four types of
DESs, may inspire scientists, who work in organic field to glimpse the potential of DESs. So
far, from the study of many reactions, it was observed that reaction in DES along with
additional greener methods like microwave irradiation or ultrasonication also helps to increase
efficiency and productivity of reactions. It is observed that viscosity plays a very important role
in DESs to act as organic solvent. Miracle of DESs in many organic reactions is its recyclability
and its reuse, without loss of its activity many times without formation of side products which
may affect yield of pure product. Literature study reveals, increase in product yield, higher
reaction rates, and reduce reaction time and no need of further purification of product using
thin layer chromatography, as compared with the conventional organic synthetic route. On
comparison of four types of reactions in DESs media, DESs Type III is mostly used in many
organic transformations, while DES Type II is very rarely used as reaction media. Thus still

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further research is needed to study mechanism involved in organic transformation involving

DESs as reaction solvent. From environmental point of view in an area of organic chemistry,
applications of DESs as reaction media, is getting more importance every day and their
viewpoint in both laboratory and industry may opens a new era in organic synthesis. But beside
of enormous green applications of DESs the next step will be focused on study of its toxicity.
References
[1] Poliakoff M & Licence P, Nature, 450 (2007) 810.
[2] Amelio A, Genduso G, Vreysen S, Luis P & Van der Bruggen B, Green Chem., 16
(2014) 3045.
[3] Zhang X, Heinonen S & Levänen E, RSC Adv., 4 (2014) 61137.
[4] Marcus Y, Biofuels and Biorefineries, Fang Zh, Xu Ch (Eds) (Volume 2||) (Springer:
Dordrecht) (2014) p.19.
[5] Marcus Y, Ionic Liquid Properties (Springer Intl. Publ., Switzerland) (2016) p.39.
[6] Abbott A P, Capper G, Davies D L, Rasheed R K & Tambyrajah V, Chemical
Communications, 1 (2002) 70.
[7] Xie Y, Dong H, Zhang S, Lu X & Ji X, Green Energy & Environment, 1 (2016) 195.
[8] García-Álvarez J, Technologies for the Environment, (Volume 1186||) (ACS
Symposium Series; American Chemical Society: Washington, DC) (2014), p.39.
[9] Francisco M, González A S B, García de Dios S L, Weggemans W & Kroon M C, RSC
Adv., 3 (2013) 23553.
[10] Abbott A P, Boothby D, Capper G, Davies D L & Rasheed R K, Journal of the
American Chemical Society 126 (2004) 9142.
[11] Seddon K R, ChemInform, 28 (2010).
[12] Smith E L, Abbott A P & Ryder K S, Chem. Rev., 114 (2014) 11060.
[13] Martins M A R, Pinho S P & Coutinho J A P, J. Solut. Chem., 48 (2018) 962.
[14] Marcus Y, Deep eutectic solvents (e-book) Springer, Cham, (Springer Nature:
Switzerland AG) (2019) p.17.
[15] Francisco M, Bruinhorst van der A & Kroon C M, Green Chemistry, 14 (2012) 2153.
[16] Francisco M, Bruinhorst van der A & Kroon C M, Angewandte Chemie, 52 (2013)
3074.
[17] Abbott A P, Capper G, Davies D L, Munro H L, Rasheed R K & Tambyrajah V,
Chem. Commun., 1 (2001) 2010.
[18] Nguyen V T, Nguyen H T & Tran P H, New Journal of Chemistry, 45 (2021) 2053.
[19] Zhang L, Hu Z, Chen X, Yan L, Liu Y & Xie Z,. Chinese Journal of Organic
Chemistry, 41 (2021) 4415.
[20] Xiao L, Liu G, Li Z, Ren P, Ren L & Kong J, (2020). Chinese Journal of Organic
Chemistry, 40 (2020) 2988.
[21] Chen G, Xie Z, Liu Y, Meng J & Zhang L..Chinese Journal of Organic Chemistry, 40
(2020) 156.
[22] Shahbaz K, AlNashef I M, Lin R J T, Hashim M A, Mjalli E S & Farid M M, Solar
Energy Mater Solar Cell, 155 (2016) 147.
[23] Chen B, Li Z, Feng Y, Hao W, Sun Y, Tang X, Zeng X & Lin L, ChemSusChem, 14
(2021) 847.
[24] Mjalli F S, Naser J, Jibril B, Alizadeh V & Gano Z, J Chem Eng Data, 59 (2014)
2242.
[25] Asadi H, Anaraki‐Ardakani H, Torabi P & Taheri N, Revue Roumaine De Chimie, 65
(2020) 795.
[26] Goudarzi H, Habibi D & Monem A, Scientific Reports, 13 (2023).
[27] Komar M, Kraljević T G, Jerković I & Molnar M, Molecules, 27 (2022) 558.

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[28] Khoshdel M A, Shirini F, Langarudi M S N, Zabihzadeh M & Biglari M, New Journal

of Chemistry, 45 (2021) 3138.
[29] Singh R R, Singh T P, Devi T L, Devi T J & Singh O M, Current Research in Green
and Sustainable Chemistry, 4 (2021) 100130.
[30] Devi T J, Singh T K, Singh R R, Singh E H & Singh O M, ChemistrySelect, 5 (2020)
13351.
[31] Biglari M, Shirini F, Mahmoodi N O, Zabihzadeh M, Langarudi M S N & Khoshdel
M A, Polycyclic Aromatic Compounds, 42 (2020) 1452.
[32] Abbott A P, Barron J C, Ryder K S & Wilson D, Chem. Eur. J., 13 (2007) 6495.
[33] Shaibuna M, Hiba K & Theresa L V, New Journal of Chemistry, 44 (2020) 14723.
[34] Dutta A, Garg A, Borah J, Borah R P & Sarma D, Current Research in Green and
Sustainable Chemistry, 4 (2021) 100107.
[35] Zwain A A, Ahmad I, Khalaf Jebur Ali R, Kahtan M, Khdyair Hamad A, Abdulgader
Hassan E, Asiri M, Ridha B M and Alsalamy A, Front. Mater., 10 (2023) 1196583.
[36] Machingal S & Krishnapillai S, Beilstein Arch, (2020) 202097.

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58

Synthesis, Characterization and Pharmacological Evaluation of Some New

Functionalized Flavone Derivatives From Β-Diketones

Sushil K. Pagariya* and Pravin S. Bodkhe

Post Graduate Department of Chemistry, Vidya Bharati Mahavidyalaya, Amravati 444602, India.
*Corresponding author email: [email protected]

ABSTRACT: In present work, a series of novel flavone analogues have been synthesized by
dehydrative cyclization of 1,3-diketone (β-diketone) derivatives carrying p-chloro-m-cresol
moiety under acidic condition and characterized by usual chemical characteristics, elemental
analysis, IR, H NMR and Mass spectral data studies for structure assignment. All the newly
1

synthesized compounds were screened for their in vitro antimicrobial activities against human
pathogenic bacterial and fungal strains by disc diffusion method. Moreover, they have been
evaluated for their in vitro antioxidant and anti-inflammatory properties by DPPH free radical
scavenging and inhibition of protein denaturation methods respectively with reference to
standard drugs.
Keywords: Flavone, β-diketone, p-chloro-m-cresol, synthesis, pharmacological evaluation.

INTRODUCTION
Flavonoids (from the Latin word flavus, which means ‘yellow’) are a large group of
natural polyphenolic compounds with low-molecular weight; particularly, they belong to a
class of plants secondary metabolites widely found in fruits, vegetables and plants derived
beverages such as green tea and wine. Flavonoids are associated with a broad spectrum of
health-promoting effects and are an indispensable component in a variety of nutraceuticals,
pharmaceuticals, medicinal and cosmetic applications because of their oxidative, anti-
inflammatory, anti-mutagenic and anti-carcinogenic properties coupled with their capacity to
modulate key cellular enzyme functions. Structurally, flavonoids is 2-phenyl benzo-γ-pyrone
(2-phenyl chromone) consists of fifteen basic carbon atoms (C6–C3–C6) with a general
formula C15H10O2 and classified into six subclasses on their chemical structure1 among which
flavone is the most prominent class of flavonoids based on the backbone of 2-phenyl-4H-
chromen-4-one (2-phenyl-1-benzopyran-4-one). Flavones are heterocyclic polyphenolic
compounds ubiquitously present in plant kingdom, especially in seeds, citrus fruits, olive oil,
tea and red wine, vegetables, nuts, stems and flowers, honey and are commonly consumed with
the human diet. Flavones display a broad spectrum of biological properties, including
antibacterial2, antifungal3, antiviral4, antidiabetic5, anti-inflammatory6-7, antioxidant8-9,
hepatoprotective10, anticancer11, anti-tubercular12, antitumoral13, and antimalarial14 activities,
making them an attractive target for synthesis and further study. Number of methods have been
developed for the synthesis of flavones and its derivatives15-17, among them Claisen-Schmidt
condensation and Baker-Venkataraman rearrangement (BVT) are traditional methods to made
flavones18. Under Claisen-Schmidt conditions, hydroxychalcones produced from 2-hydroxy-
acetophenone and benzaldehyde can undergo oxidative cyclization to yield flavone rings. In
Baker-Venkataraman approach19-20, 2-hydroxyacetophenones are converted into benzoyl
esters, which are rearranged by a base to a 1,3-diphenylpropane-1,3-diones (which exist also
in equilibrium with their enolic forms) followed by cyclization under acidic condition to yield
flavones.
In view of pharmacological significance of flavones and in continuation of our work on
synthesis of derivatives of β-diketone, it was decided to prepare some novel flavone analogues

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through cyclodehydration of 1,3-diketones (β-diketones) and to study their pharmacological

properties. Initially, the 5-chloro-2-hydroxy-4-methylacetophenone (2) was prepared by an
acetylation of p-chloro-m-cresol (a) followed by Fries rearrangement of p-chloro-m-cresyl
acetate (1) with anhydrous AlCl3. Then 2-benzoyloxyacetophenones (3a-d) were synthesized
by condensation of 5-chloro-2-hydroxy-4-methylacetophenone (2) with appropriate aromatic
carboxylic acids in pyridine using POCl3. The required starting materials were prepared via
Baker-Venkataraman transformation (BVT) wherein 2-benzoyloxyacetophenones (3a-d) were
treated with base (KOH/pyridine) to form β-diketones namely 1-(5’-Chloro-2’-hydroxy-4’-
methylphenyl)-3-(substituted phenyl) propane-1,3-diones (4a-d)21. In this, all the synthesized
diketones containing p-chloro-m-cresol moiety on dehydrative cyclization in glacial acetic acid
and H2SO4 to yield the titled compounds (5a-d). The success of the synthesis and constitutions
of synthesized compounds have been confirmed through their melting point, TLC, elemental
analysis, IR, 1H NMR and Mass spectral studies. Moreover, they have been evaluated for their
antimicrobial, antioxidant and anti-inflammatory properties by known literature methods.

MATERIALS AND METHODS

All the chemicals used were of synthetic grade. Melting points were taken in open glass
capillaries and were uncorrected. All the compounds were purified by recrystallization and
their purity was monitored by TLC using Silica gel (G) plates. Elemental analyses were carried
out with a Thermo Scientific (Model FLASH 2000) instrument. IR spectra were recorded on
Shimadzu (Model IR Afinity-1CE) spectrophotometer using KBr pellets. A Bruker (Model
Avance Neo) FTNMR 500 MHz spectrometer was used to acquire 1H NMR spectra with
DMSO-d6 as the solvent and TMS as an internal fashionable. Meanwhile, mass spectra were
recorded on Waters Micromass (Model Alliance 2795 Q-TOF) spectrometer.

General procedure for the synthesis of flavone derivatives (5a-d)22

To a solution of 1-(5’-Chloro-2’-hydroxy-4’-methylphenyl)-3-(substituted phenyl)
propane-1,3-diones (4a-d) (0.025 M) in glacial acetic acid (30 ml), sulphuric acid (5 ml) was
added. The content of reaction mixture was refluxed on water bath for 2 hr followed with
occasional stirring. The reaction mixture was allowed to cooled at room temperature and
poured into crushed ice to precipitate the product. The separated product was filtered, washed
with water followed by sodium hydrogen carbonate (10%) solution and then with sufficient
cold water until the washings were neutral to litmus. The dried product was recrystallized from
hot ethanol to get shiny yellow crystals of flavones (5a-d) as desired products (Figure-1).
R2 R1
H3C OH R2 H3C O
CH3COOH
Cl C CH2 C R1 Cl
Conc. H2SO4 O
O O

(4a-d) (5a-d)

Where, 4a, 5a : R1= NO2 ; R2= H 4c, 5c : R1= Cl ; R2= H

4b, 5b : R1= Br ; R2= H 4d, 5d : R1= Cl ; R2= Cl

Figure-1: Scheme for the synthesis of the flavone derivatives (5a-d)

Pharmacological Evaluation
Antimicrobial activity

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All the newly synthesized flavones (5a-d) were screened for their in vitro antimicrobial
sensitivity against Escherichia coli (gram -ve), Staphylococcus aureus (gram +ve) bacterial
strains and Aspergillus niger, Candida albicans fungal strains by disc diffusion method22-25.
Mueller-Hinton agar (MHA) and Potato Dextrose agar (PDA) plates were employed as culture
medium respectively for bacterial and fungal sensitivity and DMSO was used as solvent
control. Ofloxacin and amphotericin were used as standard drugs for antibacterial and
antifungal activities respectively. The compounds were dissolved in DMSO to give 100 µg/ml,
250 µg/ml, 500 µg/ml solutions. Sterile filter paper discs (Whatmann filter paper No. 40) of 10
mm diameter were dipped in these solutions, dried, and placed on nutrient agar plates spreaded
with the bacteria and fungi. The plates were further incubated for 24 hr at 37 0C for antibacterial
and 72 hr at 28 0C for antifungal testing and the zones of inhibition were measured in mm using
antibiotic zone reader (Hi-Media).

Antioxidant activity
The in vitro antioxidant activity of synthesized compounds was performed based on
DPPH radical-scavenging assay with free radical scavenging effect of the stable 2, 2-diphenyl-
1-picrylhydrazyl (DPPH) with slight modifications26-29. A stock solution of DPPH (1.3 mg/ml)
was prepared by dissolving 13.0 mg in 10 ml of methanol. The stock solutions of test
compounds (5a-d) (100 μg/ml) were prepared and further diluted to prepare solutions of various
concentrations (10, 20, 30, 40 and 50 μg/ml). Similarly, a stock solution of standard ascorbic
acid (1 mg/ml) was prepared by dissolving 10 mg ascorbic acid in 10 ml of methanol. From
this stock solution further dilutions of various concentrations (10, 20, 30, 40, and 50 μg/ml)
were prepared. Further, 1 ml of DPPH solution was mixed with 1 ml of different concentration
of test solutions and standard (ascorbic acid). These solutions were kept for 30 min in dark and
absorbance was measured at 517 nm using methanol (5 ml) with DPPH solution (1 ml) as blank
(positive control). The percentage of scavenging activity was calculated by following formula:
% Scavenging = [Abs. of control–Abs. of test sample/Abs. of control] × 100

Anti-Inflammatory activity
The in vitro anti-inflammatory activity of synthesized compounds was performed by
employing inhibition of protein denaturation assay30. Stock solution of NSAID ibuprofen as a
reference drug (positive control) was prepared by dissolving 10 mg of ibuprofen in 10 ml of
distilled water. Serial dilution from above stock solution takes 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml,
0.5 ml and prepare 10 ppm, 20 ppm, 30 ppm, 40 ppm, 50 ppm and also it was performed for
four sample extract (5a-d). All samples contain 5 ml of total volume. Reaction mixtures were
prepared using 2.8 ml of phosphate-buffered saline (pH 6.4) and 0.2 ml of egg albumin. Then
take 2 ml of extract from each different concentration were mixed gently with reaction
mixtures. A similar procedure was used for reference drug ibuprofen. The absorbance of these
solutions was determined by spectrophotometer at wavelength of 660 nm. The percentage
inhibition of protein denaturation was calculated by following formula:
% Inhibition = [Abs. of blank – Abs. of test sample/Abs. of blank] × 100

RESULTS AND DISCUSSION

Spectral data analysis
The structures of all the newly synthesized flavones (5a-d) were characterized on the
basis of melting point, TLC, elemental analysis, IR, 1H NMR and Mass spectral studies. The
IR, 1H NMR spectra showed expected signals corresponding to various groups present in each
of the compounds. The mass spectrum of all the compounds were found to in full agreement
with the proposed structures and showed expected peaks which confirms the molecular weights
of compounds. The spectral and physical data (Table-1) of all the compounds are shown below:

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(5a): 2-(4’-Nitrophenyl)-6-chloro-7-methyl-4H-chromen-4-one: IR (cm-1, KBr): 3085 [C-H

stretch (aromatic)], 2930 [C-H stretch in -CH3 (aliphatic)], 1647 (C=O stretch), 1525 [C=C
stretch (aromatic)], 1431 [-NO2 stretch (asym.)], 1345 [-NO2 stretch (sym.)], 1247 (C-O
stretch), 655-691 (C-Cl stretch); 1H NMR (DMSO-d6/500 MHz):  2.5 ppm (s, 3H of -CH3), 
3.38 ppm (s,1H of -C=CH- of flavone),  7.9-8.4 ppm (m, 6H of Ar-H); MS (m/z): 316.04(M+).
(5b): 2-(4’-Bromophenyl)-6-chloro-7-methyl-4H-chromen-4-one: IR (cm-1, KBr): 3046 [C-H
stretch (aromatic)], 2924 [C-H stretch in -CH3 (aliphatic)], 1632 (C=O stretch), 1434 [C=C
stretch (aromatic)], 1244 (C-O stretch), 648 (C-Br stretch); 1H NMR (DMSO-d6/ 500 MHz): 
2.5 ppm (s, 3H of -CH3),  3.4 ppm (s,1H of -C=CH- of flavone ring),  7.8-8.1 (m, 6H of Ar-
H); MS (m/z): 348.96/350.96 (M+).
(5c): 2-(4’-Chlorophenyl)-6-chloro-7-methyl-4H-chromen-4-one: IR (cm-1, KBr): 3043 [C-H
stretch (aromatic)], 2922 [C-H stretch in -CH3 (aliphatic)], 1724 (C=O stretch), 1438 [C=C
stretch (aromatic)], 1254 (C-O stretch), 725 (C-Cl stretch); 1H NMR (DMSO-d6/ 500 MHz): 
2.5 ppm (s, 3H of -CH3),  3.4 ppm (s,1H of -C=CH- of flavone ring),  7.6-8.1 ppm (m, 6H
of Ar-H); MS (m/z): 305.02 (M+).
(5d): 2-(2’,4’-Dichlorophenyl)-6-chloro-7-methyl-4H-chromen-4-one: IR (cm-1, KBr): 3284
[C-H stretch (aromatic)], 2928 [C-H stretch in -CH3 (aliphatic)], 1635 (C=O stretch), 1427
[C=C stretch (aromatic)], 1103 (C-O stretch), 628 (C-Cl stretch); 1H NMR (DMSO-d6/ 500
MHz):  2.5 ppm (s, 3H of -CH3),  3.34 ppm (s,1H of -C=CH- of flavone ring),  7.0-8.1 (m,
5H of Ar-H); MS (m/z): 338.99 (M+).

Table-1: Physical and analytical data of synthesized substituted flavones (5a-d)

Compound Molecular Mol.wt m.p. % Analysis: Found (Calcd.) Yield Rf
code Formula (g/mol) (0C) C H Cl (%) value
5a C16H10ClNO4 315.71 160-162 61.08 3.06 11.30 80 0.64
(60.87) (3.19) (11.23)
5b C16H10BrClO2 349.61 166-169 55.29 2.79 (9.86) 78 0.68
(54.97) (2.88) (10.14)
5c C16H10Cl2O2 305.16 156-158 63.67 3.79 21.99 88 0.70
(62.97) (3.30) (23.24)
5d C16H9Cl3O2 339.60 170-172 56.77 2.81 (31.16) 92 0.69
(56.59) (2.67) (31.32)
Pharmacological activity
The antibacterial and antifungal activities of all the newly synthesized compounds were
examined in vitro tests. The resulting data on antimicrobial activities are depicted in Table-2
and Table-3. They were further subjected for their in vitro antioxidant and anti-inflammatory
activity whose results are summarized respectively in Table-4 and Table-5 and also shown in
Figure-2 and Figure-3 statistically.

Table-2: Antibacterial activity of newly synthesized flavone derivatives (5a-d)

Compound Zone of inhibition in mm
code Escherichia coli (Gram -ve) Staphylococcus aureus (Gram +ve)
Concentration of compounds (μg/ml) Concentration of compounds (μg/ml)
100 250 500 Std. 100 250 500 Std.
5a NI NI NI 20 mm NI NI NI 22 mm
5b NI NI NI NI NI NI

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5c NI NI NI NI NI NI
5d 10 mm 12 mm 14 mm 10 mm 11 mm 12 mm
Control NI NI NI NI NI NI
Note: Standard: Ofloxacin (2 μg/ml); (NI): No Zone of Inhibition; Control: DMSO solvent

Table-3: Antifungal activity of newly synthesized flavone derivatives (5a-d)

Compound Zone of inhibition in mm
code Aspergillus niger Candida albicans
Concentration of compounds (μg/ml) Concentration of compounds (μg/ml)
100 250 500 Std. 100 250 500 Std.
5a NI NI NI 15 mm NI NI NI 14 mm
5b NI NI NI NI 11 mm 12 mm
5c NI 10 mm 11 mm NI 10 mm 11 mm
5d NI NI NI NI 11 mm 12 mm
Control NI NI NI NI NI NI
Note: Standard: Amphotericin (50 μg/ml); (NI): No Zone of Inhibition; Control: DMSO solvent

According to the obtained results, compounds 5a, 5b and 5c showed zero zones of
inhibition at all the tested concentrations and were found to be inactive against E. coli and S.
aureus while compound 5d showed 10, 12, 14 mm and 10, 11, 12 mm zones of inhibition
respectively at 100 μg/ml, 250 μg/ml and 500 μg/ml against E. coli and S. aureus organisms.
Standard ofloxacin (2 μg/ml) showed 20 mm and 22 mm zones of inhibition respectively
against bacteria E. coli and S. aureus. In case of antifungal sensitivity, compounds 5a, 5b and
5d showed no zones of inhibition at all the concentrations and were found to be inactive against
A. niger, while compound 5c showed 10 and 11mm of zone at 250 μg/ml and 500 μg/ml
concentrations respectively against A. niger. Standard amphotericin (50 μg/ml) showed 15 mm
of zone against A. niger. As far as fungus C. albicans is concern, compound 5b, 5c, 5d were
found to show 11, 12; 10, 11; and 11, 12 mm of zones of inhibition at 250 μg/ml and 500 μg/ml
concentrations respectively whereas compound 5a was found to be inactive against C. albicans
at all the tested concentrations when compared with std. amphotericin drug which showed 14
mm of zone against C. albicans.

Table-4: Antioxidant activity of newly synthesized flavone derivatives (5a-d)

Compd. code % Scavenging of synthesized compounds (5a-d) and std. Ascorbic acid
10 μg/ml 20 μg/ml 30 μg/ml 40 μg/ml 50 μg/ml
5a 39.69 45.17 59.42 68.34 79.76
5b 19.51 23.46 24.78 30.33 38.56
5c 26.97 35.96 46.92 60.22 75.09
5d 21.05 35.96 36.84 59.70 71.00
Standard 80.26 91.22 95.61 96.27 97.80

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Table-5: Anti-inflammatory activity of newly synthesized flavone derivatives (5a-d)

Compd. code % Inhibition of synthesized compounds (5a-d) and std. Ibuprofen drug
10 μg/ml 20 μg/ml 30 μg/ml 40 μg/ml 50 μg/ml
5a 31.51 59.71 66.72 76.85 85.43
5b 51.65 52.81 54.30 59.95 67.76
5c 39.18 40.17 53.09 60.41 74.38
5d 47.67 52.81 66.39 73.11 81.67
Standard 87.36 88.90 90.23 91.74 93.10

The experimental data on antioxidant activities (Table-4) reveals that, compounds 5a,
5c and 5d showed good scavenging activity while compound 5b showed mild scavenging
activity with reference to std. ascorbic acid. Also, one thing is noted that, as the concentration
of test compounds increases, their percentage of scavenging also increases. In case of anti-
inflammatory activities, we observed that, compounds 5a and 5d showed excellent inhibition
to protein denaturation as compared to compound 5b and 5c which showed moderate to good
inhibition to protein denaturation with comparison to std. NSAID ibuprofen drug.
10 μg/ml 20 μg/ml 30 μg/ml 40 μg/ml 50 μg/ml
10 μg/ml 20 μg/ml 30 μg/ml 40 μg/ml 50 μg/ml
% Scavenging of test compounds

100
90
% Inhibition of test compounds

80 100
70 90
60 80
50 70
40 60
30 50
20 40
10 30
0 20
10
Standard 5a 5b 5c 5d 0
Standard 5a 5b 5c 5d
Figure-2: Graph showing percentage of Figure-3: Graph showing percentage
scavenging of test compounds (5a-d) of inhibition of test compounds (5a-d)

CONCLUSION
In conclusion, a total four novel flavones viz. 2-(substituted phenyl)-6-chloro-7-
methyl-4H-chromen-4-one (5a-d) were successfully synthesized by dehydrative cyclization of
β-diketones bearing moiety of p-chloro-m-cresol and characterized by spectral studies. All the
synthesized compounds were evaluated for their antimicrobial, antioxidant and anti-
inflammatory potential in vitro. From the results, it was clear that, only the compound 5d was
moderately active while compounds 5a, 5b and 5c were found to be inactive against E. coli and
S. aureus as compared to std. antibiotic ofloxacin. Also, it was found that, only the compound
5c was moderately active against A. niger while compounds 5a, 5b, 5d do not shown inhibitory
action and were found to inactive against A. niger when compared with std. amphotericin drug.
As far as C. albicans is concern, instead of compound 5a, the compounds 5b, 5c, and 5d were
found to show moderate to good inhibitory action to growth response of fungus C. albicans.
The results on antioxidant and anti-inflammatory activities reveals that, among the newer
derivatives, compounds 5a, 5c, 5d showed a promising antioxidant activity as well as all the
compounds showed notable and excellent anti-inflammatory activities with comparison to std.
reference drugs. It is conceivable that these newly synthesized flavones showing in vitro
scavenging and protein denaturation activity can be further modified to achieve marketable
antioxidant and NSAID agents.

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ACKNOWLEDGEMENTS
The authors are thankful to Principal, Vidya Bharati Mahavidyalaya, Amravati for
providing expensive laboratory facilities. The authors are also grateful to Prof. Dr. S. L. Deore,
Govt. College of Pharmacy, Amravati and SAIF, Panjab University, Chandigarh for their help.

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59

Design, Synthesis, Spectral Characterisation and Antibacterial

Screening of Some Novel 4- Substitutedimino-1,3,5-Dithiazine
Along With Pyrimidine Nucleus

S. B. Sarkatea*, S. A. Waghmareb, K. U. Dongarec, R. N. Ingoled

a,b,c-
Department of Chemistry, Ghulam Nabi Azad Arts, Commerce and Science College Barshitakli Dist. Akola
444401 (M.S.), India.
d-
Department of Chemistry, Shri Vitthal Rukhmini College, Sawana,Mahagaon, Dist.Yavatmal,(M.S.)
India
*Corresponding author- [email protected]

ABSTRACT
In recent times in the laboratory, synthesis of 2-(2-Phenylimino-4- substitutedimino-
1,3,5-dithiazino) aminopyrimidine (IIIa-e) were synthesized by refluxing 2-(5-phenyl-,4-
dithiobiureto) pyrimidine (I) with alkyl/ arylisocyanodichlorides (IIa-e) in acetone-
ethanol medium in 1:1 molar proportion. The structures of all the synthesized compounds
were acceptable on the basis of chemical characteristics, elemental analysis, spectral studies
and their antibacterial screening against the gram positive and gram negative bacteria such as
S. typhi, E. coli, S. aureus, A. Aerogenes, B. Subtilis and B. Megatherium.etc.

INTRODUCTION
Heterocycles containing organic molecules are more fascinating, because they are
convenient in a variety of applications, a multitude of uses are included by the heterocycles'
size and heteroatom diversity. These compound nitrogen, oxygen and sulphur are composed
of six member1-5, five member6-7 fused heterocycles with aromatic rings. Particularly
noteworthy are the numerous biological and industrial uses for heterocycles that combine
sulphur and nitrogen in one ring. Six member two sulphur and one nitrogen atom prepare 1,3,5-
dithiazine. It is one of the six member heterocycles that functions as a strong medication in the
sectors of medicine, agriculture, and industry9-8.
Tayade10, Pathe11 and Mur12 Synthesized numerous heterocycles containing 1,3,5-
dithiazines as main nucleus. Each 1,3,5-dithiazino moiety has different applications
according to the substituent attached to the basic nucleus of the 1,3,5-
ithiazine. It has been also observed during literature study that, 1,3,5-dithiazino nucleus and
its derivatives possesses biological and medicinal effective properties13-14. Substituted
isocyanodichlorides were used by researchers to synthesise 1,3,5- dithiazines in the
laboratory. This 1,3,5-dithiazine synthesis technique is quicker, less complicated, less
expensive and requires less time.

MATERIAL AND METHOD

Material
All the chemical used were of loba chemie (AR grade).
Method
In the present experiment for the synthesis of different substituted 1,3,5-dithiazino

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aminopyrimidine is conventional refluxing under electronic water bath for different hours for
different experiment.
EXPERIMENTAL
All the chemicals used for the synthesis were purified. After refluxing the purity of the
compounds were checked by TLC (aluminium TLC) with thin layer thickness of 200 um. The
melting points of all synthesize compounds will be recorded using hot paraffin bath. The
carbon and hydrogen analysis were carried out on Carlo-Ebra-1106 analyser Nitrogen
estimation were carried out with colmon-N-analyzer-29. IR spectra were recorded with
Bruker spectrometer in the range 4000-400 cm-1. PMR spectra were recorded on VARIAN
400 MHz spectrometer with TMS as internal standard using CDCL3 and DMSO Solvent.
GENERAL PROCEDURE
A reaction of 2-(5-phenyl-2,4-dithiobiureto)pyrimidine (I) and
substitutedisocyanodichloride (IIa-e) in 1:1 molar ratio refluxed on water bath in acetone-
ethanol medium for 2 hours. The evolution of the hydrochloride gas was clearly observed during
refluxtion. After distillation of excess solvent orange product isolated which on basification
with dilute ammonium hydroxide orange crystalline products obtained.
Similar, procedure was adopted for the synthesis of all the derivatives in the
series.

The tentative reaction for the formation of product is depicted below,

Similarly, 2-(5-phenyl-2,4-dithiobiureto)pyrimidine (I) were react with

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phenylisocyanochloride (IIa), ethylisocyanodichloride (IIb), tertbutylisocyanodichloride (IIc), P-

tolylisocyanodichloride (IId) and 4-chlorophenhylisocyanodichloride (IIe) by the above mentioned
method to isolate 2-[2,4-di(phenylimino)-1,3,5-dithiazino] aminopyrimidine (IIIa), 2- (2-phenylimino-4-
ethylimino-1,3,5-dithiazino) aminopyrimidine(IIIb), 2-(2-phenylimino-4- tert-butylimino-1,3,5-
dithiazino) aminopyrimidine (IIIc), 2-[2-phenylimino-4-(4- methylphenylimino)-1,3,5-
dithiazino]aminopyrimidine (IIId) and 2-[2-phenylimin-4-(4- chlorophenylimino)-1,3,5-
dithiazino]aminopyrimidine (IIIe).

RESULT AND DISCUSSION

Spectral characterization results for all the synthesized compounds are given below
Spectral Characterization
1. 2-(2,4-diphenylimino-1,3,5-dithiazino)aminopyrimidine (IIIa)
Colour-Yellow solid, Molecular formula- C15H14N6S2, Yield 85%, M.P. 1620C, %
Composition found (calculated) C-52.90 , H-4.83 , N-25.20, S-23.21 , FTIR (Kbr)
vcm- 3245.22 N-H stretching, 3051.47 (C-H Ar Stretching), 3187.38 (N-H Amido),
1950.37 (C-H Ar Bending,), 1185.99 (C=S Stretching), 670.04(=C-H bending); 1H
NMR (400MHz CDCL3 δ ppm), 8.4ppm (2H, double, CH) of pyrimidine, 7.2ppm (2H,
CH, doublet) of pyrimidine, 7.2 ppm ( 2H, , doublet CH) of phenyl, 3.4 ppm (1H, singlet
NH), 4.6 ppm 1H singlet, 7.39 1H triplet CH benzene, 1H triplet CH benzene
2.2 ppm 2H quartet, 2.3 ppm 3H triplet Mass m/z 390.30
2. 2-(2-phenylimino-4-ethylimino-1,3,5-dithiazino)aminopyrimidine (IIIb)
Colour-Yellow solid, Molecular formula- C15H14N6S2, Yield 86%, M.P. 1700C, %
Composition found (calculated) C-52.90 , H-4.83 , N-25.20, S-23.21 , FTIR (Kbr)
vcm- 3245.22 N-H stretching, 3051.47 (C-H Ar Stretching), 3187.38 (N-H Amido),
1950.37 (C-H Ar Bending,), 1185.99 (C=S Stretching), 670.04(=C-H bending); 1H
NMR (400MHz CDCL3 δ ppm), 8.4ppm (2H, double, CH) of pyrimidine, 7.2ppm (2H,
CH, doublet), 7.2 ppm ( 2H, , doublet CH) of phenyl, 3.4 ppm (1H, singlet NH),
4.6 ppm 1H singlet, 7.39 1H triplet CH benzene, 1H triplet CH benzene 2.2 ppm 2H quartet,
2.3 ppm 3H triplet Mass m/z 342.30
3. 2-(2-phenylimino-4-tertbutylimino-1,3,5-dithiazino)aminopyrimidine (IIIc)
Colour-Yellow solid, Molecular formula- C17H18N6S2, Yield 90%, M.P. 1580C, %
Composition found (calculated) C-55.90 , H-4.83 , N-23.20, S-17.21 , FTIR (Kbr)
vcm- 3245.22 N-H stretching, 3045.47 (C-H Ar Stretching), 3140.38 (N-H Amido),
1995.37 (C-H Ar Bending,), 1164.99 (C=S Stretching), 710.04(=C-H bending); 1H
NMR (400MHz CDCL3 δ ppm), 8.2ppm (2H, double, CH), 7.2ppm (2H, CH,
doublet), 7.1ppm ( 2H, , doublet CH), 3.5 ppm (1H, singlet NH), 4.5 ppm 1H singlet,
7.40 1H triplet CH benzene, 2.4 9H singlet Mass m/z 370.50
4. 2-(2-phenylimino-4-tolylimino-1,3,5-dithiazino)aminopyrimidine (IIId)
Colour-Yellow solid, Molecular formula- C20H16N6S2, Yield 82%, M.P. 1670C, %
Composition found (calculated) C-55.90 , H-4.83 , N-23.20, S-15.21 , FTIR (Kbr)
vcm- 3200.22 N-H stretching, 2901.47 (C-H Ar Stretching), 316038 (N-H Amido),
1960.37 (C-H Ar Bending,), 1167.99 (C=S Stretching), 745.04(=C-H bending); 1H
NMR (400MHz CDCL3 δ ppm), 8.2 ppm (2H, double, CH) of pyrimidine, 7.3ppm (2H,
CH, doublet), 7.2 ppm ( 2H, , doublet CH), 3.5 ppm (1H, singlet NH), 4.7 ppm 1H singlet,
7.39 1H triplet CH benzene, 3.4 ppm 3H singlet. Mass m/z 404.60

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5. 2-(2-phenylimino-4-p-chlorophenylimino-1,3,5-dithiazino)aminopyrimidine
(IIIe)
Colour-Yellow solid, Molecular formula- C19H13N6S2, Yield 80%, M.P. 1450C, %
Composition found (calculated) C-53.90 , H-3.83 , N-20.20, S-15.21Cl-8.45 , FTIR
(Kbr) vcm- 3263.22 N-H stretching, 3051.47 (C-H Ar Stretching), 3216.38 (N-H
Amido), 1966.37 (C-H Ar Bending,), 1165.99 (C=S Stretching), 720.04(=C-H bending);
1H NMR (400MHz CDCL3 δ ppm), 8.4 ppm (2H, double, CH) of pyrimidine,
7.03ppm (2H, CH, doublet), 7.4 ppm ( 2H, , doublet CH), 3.5 ppm (1H, singlet NH), 4.6 ppm 1H
singlet, 7.39 1H triplet CH benzene, Mass m/z 424.80.

CONCLUSION
In the present work is cheaper and less time consuming method for synthesis of
organic compound (IIIa-e). In all the synthesized compounds give the maximum yield of
product (III a-e). A variety of pyrimidine based 1,3,5-dithiazine derivative can be
synthesized for their antimicrobial activities adopting the method.
PHARMACOLOGICAL STUDIES
Antimicrobial activity
All the synthesized compounds (III-a) to (III-e) were screened for antibacterial
activity against S. typhi, E. coli, S. aureus, A. Aerogenes, B. Subtilis and B. Megatherium. by
disc diffusion method was performed using mueller hinton agar as well as nutrient agar
medium. Each and every compound was tested at conc. 50 μg/ml in ethanol. The zone of
inhibition of all the synthesized compounds were measured after 24 hour incubation at 37 0C.
Standard drug used for comparison the activity was Ciprofloxacin.
Table 1: Synthesized compound IIIa-e activity against Gram +ve and Gram -ve bacteria

Table No. 1.1 - Antibacterial activity of synthesized compound against

bacteria (Zone of inhibition in mm) (after 24 hrs at 37 oC temp)

Gram positive Gram negative

SR B. A.
Compd. S. B. megatheriu S. E. aerogene
No.
aureus subtilis m typhi coli s

1 SBS-IIIa 18 12 16 14 18 11
2 SBS-IIIb 11 12 14 14 11 12
3 SBS-IIIc 11 11 12 11 11 11
4 SBS-IIId 10 12 12 12 11 11
5 SBS-IIIe 14 12 14 12 16 12
Std
6 18 16 20 18 20 14
Ciprofloxacin

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REFERANCE

1. Zhang, D. H., Zhang, Z., & Shi, M. (2012). Transition metal-catalyzed

carbocyclization of nitrogen and oxygen-tethered 1, n-enynes and diynes:
synthesis of five or six-membered heterocyclic compounds. Chemical
Communications, 48(83), 10271-10279.
2. Larrosa, I., Romea, P., & Urpí, F. (2008). Synthesis of six-membered oxygenated
heterocycles through carbon-oxygen bond-forming reactions. Tetrahedron,
64(12), 2683-2724.
3. Sanu, M. C., Joseph, J., Chacko, D., Vinod, B., & Daisy P., A. (2021). Review
on six membered nitrogen containing heterocyclic compounds with various
biological activities. International Journal of Pharmaceutical Sciences Review
and Research, 69(2), 64-68.
4. Walton, J. C. (2016). Synthetic strategies for 5-and 6-membered ring
azaheterocycles facilitated by iminyl radicals. Molecules, 21(5), 660.
5. Huh, D. N., Cheng, Y., Frye, C. W., Egger, D. T., & Tonks, I. A. (2021).
Multicomponent syntheses of 5-and 6-membered aromatic heterocycles using
group 4–8 transition metal catalysts. Chemical Science, 12(28), 9574-9590.
6. Sergey P., Vladimir N., Andrey, E., & A. Pimerzin, E. Vishnevskaya E.,
Thermodynamic analysis of strain in the five-membered oxygen and nitrogen
heterocyclic compounds. The Journal of Physical Chemistry, 115( 10), 1992–
2004.
7. Kaur, N., Yadav, N., & Verma, Y. (2023). Acetamidine in heterocycle synthesis.
Synthetic Communications, 53(9), 577-614.
8. Gujjar, K. N., & Narasimha S M., (2023).A Review: Important applications of
Heterocyclic Compounds. Europeam Chemical Bulletin, 12(12), 625-630.
9. Sharma, P. K., Amin, A., & Kumar, M. (2020). A review: Medicinally important
nitrogen sulphur containing heterocycles. The Open Medicinal Chemistry
Journal, 14(1), 49-64.
10. Tayade, D. T., & Padhen, S. S. (2016). Synthesis and Characterization of 1-
Phenyl-3- [4-(2- Substitutedimino-4-Substitutedimino-1,3,5-
Dithiazino) Aminophenyl]- Prop-2-Ene-1-Ones. International Journal of
Pharmacy and Pharmaceutical Research, 7 (1), 53-58.
11. Pathe, P. P., & Paranjpe, M. G. (1984). Preparation of 5‐aryl‐4‐ arylimino‐6‐
benzylimino‐2‐ phenylimino‐1,3,5‐dithiazine. Indian
chemical society, 15(43), 149-150.
12. Mur, V. I. (1964). 2, 4, 6-trichloro-1, 3, 5-triazine (cyanuryl chloride) and its
future applications. Russian Chemical Reviews, 33(2), 92-103.
13. Panpaliya, K. S., Tayade, D. T., Shaikh, R. S., & Thakare, A. N. (2017). Synthesis
of 1-phenyl-3-substituted- 2,6-dithio-4- amino-[(2- phenylthiocarbamido)- 1,3-
benzothiazolo] -1,3,5-triazine and their effects on germination pattern of
sorghum vulgare. Online International Interdisciplinary Research Journal, 1, 6-
9.
14. Deohate, P. P., & Berad, B. N. (2005). Synthesis and antimicrobial activity of 1,
3, 5-thiadiazines and their isomerism into 1, 3, 5-triazines. Indian journal of
Chemistry, 44B, 638-642.

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60

Colorimetric Phytochemical Analysis of Tinospora Cordifolia and FTIR

Study of Its Medicinal Sample Giloy Ghanvati
Dr Swaroopa Rani N. Gupta
Professor, Department of Chemistry
Brijlal Biyani Science College, Amravati, Maharashtra India
[email protected]

ABSTRACT
Tinospora cordifolia (common names gurjo, heart-leaved moonseed, guduchi or giloy) is a
herbaceous vine of the family Menispermaceae indigenous to tropical regions of the Indian
subcontinent. It has been used in Ayurveda to treat various disorders. Tinospora cordifolia
contains diverse phytochemicals, including alkaloids, phytosterols, glycosides, tinosporide,
and other mixed chemical compounds. During the 2020-22 COVID-19 outbreak in India, the
Ministry of AYUSH recommended use of Giloy as a home remedy for immune support. Giloy
extract was assessed for its phytochemical and antibacterial properties, to find out the bioactive
components responsible for such activity.

In Ayurveda, it is said to be the best rejuvenating herb. Acute toxicity studies of aqueous extract
of Guduchi reports that it does not produce any toxic effect. The medicinal herb has to be used
in an appropriate dose as prescribed by a qualified physician to get medicinal effects. With the
wide range of actions and abundant components, Guduchi is a real treasure among herbal drug
source. Medicinal applications of Guduchi in countering various disorders and it's use as anti-
oxidant, anti-hyperglycemic, anti-hyperlipidemic, hepatoprotective, cardiovascular protective,
neuroprotective, osteoprotective, radioprotective, anti-anxiety, adaptogenic, analgesic, anti-
inflammatory, anti-pyretic, anti-diarrheal, anti-ulcer, anti-microbial, and anti-cancer have been
well established.

Present study deals with Colorimetric phytochemical analysis of Tinospora Cordifolia (Giloy
leaf extract) and FTIR study of its medicinal sample Giloy Ghanvati. Colorimetric
phytochemical analysis of Tinospora Cordifolia (Giloy leaf extract) involves preparation of
standard Giloy Ghanvati solution, Ferric Chloride solution and Giloy leaf Solution. Different
systems were prepared. Absorbance of standard Giloy solution was taken at different
wavelength. And λmax was calculated. Which was 420 nm. Absorbance of standard Giloy
solution of different concentrations were taken at 420 nm and Calibration plot was prepared.
Then absorbance of Giloy leaf solutions were taken at 420 nm and concentration of Giloy in
Giloy leaf was calculated from calibration plot. Tinospora cordifolia which contains diverse
phytochemicals, including alkaloids, phytosterols, glycosides, tinosporide, and other mixed
chemical compounds reduces ferric ions and production of violet coloration by addition of
ferric chloride to dilute solution of Giloy takes place and the concentration of Giloy was
investigated colorimetrically. The method is simple, rapid and precise. Interpretation of FTIR
Spectra of Giloy Ghanvati medicinal sample shows presence of various functional groups such
as O-H stretching - Alcohol, Carboxylic acid, O-H bending - Alcohol, N-H stretching - Amine
salt, C=C stretching - α, β- unsaturated ketone and C=O stretching - Conjugated acid.

Keywords: Tinospora cordifolia, Giloy, Giloy ghanvati, Colorimetric Phytochemical

Analysis, FTIR study

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INTRODUCTION

Giloy
Tinospora cordifolia (common names gurjo, heart-leaved moonseed, guduchi or giloy) is a
herbaceous vine of the family Menispermaceae indigenous to tropical regions of the Indian
subcontinent. It has been used in Ayurveda to treat various disorders, but in spite of clinical
investigation, the effectiveness of such treatments remains uncertain. [1] Reddish fruits of
Tinospora cordifolia is a large, deciduous, extensively-spreading, climbing vine with several
elongated twining branches.

Leaves of Tinospora cordifolia Fruits of Tinospora cordifolia

Endophytic fungi colonize the living, internal tissues of their host without causing any harmful
effects. A recent study has shown that 29 endophytes belonging to different taxa were present
in the samples collected from Tinospora cordifolia.[2] Extracts of the endophytic fungus
Nigrospora sphaerica obtained from T. cordifolia were found to have insecticidal properties
against the Oriental leafworm moth (Spodoptera litura), a polyphagous pest.[3] Tinospora
cordifolia contains diverse phytochemicals, including alkaloids (Alkaloids are a class of basic,
naturally occurring organic compounds that contain at least one nitrogen atom), phytosterols,
glycosides, tinosporide,[4] and other mixed chemical compounds. Although used in Ayurveda
over centuries in the belief that Tinospora has medicinal properties.[5]

Phytosterols Glycosides Tinosporide

During the 2020-22 COVID-19 outbreak in India, the Ministry of AYUSH recommended use
of T. cordifolia (Giloy) as a home remedy for immune support, [6] but such a practice appeared

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to be associated with hepatitis cases among six people in Mumbai who used boiled or capsule
preparations of the plant. [6-9]

Tinospora cordifolia is a popular medicinal plant which is used in several traditional medicines
to cure various diseases. The common names are Amrita and Guduchi and belong to the family
of Menispermaceae. It is considered an essential herbal plant of Indian system of medicine
(ISM) and has been used in the treatment offever, urinary problem, dysentery, skin diseases
leprosy, diabetes, and many more diseases. The plant reported containing chemical compound
including Alkaloids, Terpenoids, Lignans, Steroids and others that establish the
phytochemistry and pharmacological activity of Tinospora cordifolia. The study highlights the
pharma-cological importance viz antioxidant activity, antimicrobial activity, antibacterial
activity, antifungal activity, anti-diabetic activity, antistress activity, hypolipidaemic effect,
hepatic disorder, anticancer anti HIV potential, antiosteoporotic effects, antitoxic effects,
wound healing, anticomplementary activity, and immunomodulating activity, systemic
infection and Parkinson̓s disease. [10]

Major constituent of Tinospora cordifolia: terpenoid, alkaloid, lignans, steroids. [10]

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Structure of the chemical constituent of T. cordifolia. [10]

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Some of the essential constituents of T. cordifolia.

Terpenoids Tinosporide, Furanolactone diterpene, Furanolactone clerodane diterpene,
furanoid diterpene, Tinosporaside, ecdysterone makisterone and several glucosides isolated as
poly acetate, phenylpropene disaccharides cordifolioside A, B and C, cordifoliside D and E,
Tinocordioside, cordioside, palmatosides C and F, Sesquiterpene glucoside tinocordifolioside,
Sesquiterpene tinocordifolin. [11-21]
Alkaloids Tinosporine, (S), Magnoflorine, (S), Berberine, (S), Choline, (S), Jatrorrhizine, (S),
1,2-Substituted pyrrolidine(S), Alkaloids, viz. jatrorrhizine, palmatine, beberine, tembeterine,
choline. [12-26]
Lignans 3 (a, 4-dihydroxy-3-methoxybenzyl)-4-(4- hydroxy-3-methoxybenzyl), (S) [27]
Steroids Giloinsterol, (S), ß-Sitosterol, (S), 20aHydroxy ecdysone, (S). [28-31]
Others Giloin, Tinosporan acetate, Tinosporal acetate, Tinosporidine, Heptacosanol,
Octacosanol, sinapic acid, Tinosponone, two phytoecdysones, an immunologically active
arabinogalactan. [32-36]

The study planned to prepare antibacterial finish for grey cotton fabric using Giloy stem extract
for healthcare applications. The selected grey cotton fabric was pretreated prior to application
of the extract. For the extraction of the herb, maceration process was employed and the solution
prepared was further subjected to soxhlet extraction to congeal the extract. Giloy extract was
assessed for its phytochemical and antibacterial properties, to find out the bioactive
components responsible for such activity. [37]

Certain sections of the media have falsely linked again Giloy/Guduchi to liver damage. The
Ministry of Ayush reiterates that Giloy/Gudduchi (Tinospora cordifolia) is safe and as per
available data, Guduchi does not produce any toxic effect. In Ayurveda, it is said to be the best
rejuvenating herb. Acute toxicity studies of aqueous extract of Guduchi reports that it does not
produce any toxic effect. However, the safety of a drug depends on how it is being used. Dosage
is one of the important factors that determine the safety of a particular drug. In a study, lower
concentration of Guduchi powder is found to increase the life span of fruit flies (Drosophila
Melanogaster). At the same time, higher concentration progressively reduced the life span of
flies. This clearly indicates that an optimum dosage should be maintained in order to get the
desired effects. This infers that the medicinal herb has to be used in an appropriate dose as
prescribed by a qualified physician to get medicinal effects. With the wide range of actions and
abundant components, Guduchi is a real treasure among herbal drug source. Medicinal
applications of Guduchi in countering various disorders and it's use as anti-oxidant, anti-
hyperglycemic, anti-hyperlipidemic, hepatoprotective, cardiovascular protective,
neuroprotective, osteoprotective, radioprotective, anti-anxiety, adaptogenic, analgesic, anti-
inflammatory, anti-pyretic, anti-diarrheal, anti-ulcer, anti-microbial, and anti-cancer have been
well established.

A special focus has been made on its health benefits in treating various metabolic disorders and
its potential as an immune booster. It is used as a major component of therapeutics for
ameliorating metabolic, endocrinal, and several other ailments, aiding in the betterment of
human life expectancy. It is a popularly known herb for its immense therapeutic applications
in traditional systems of medicine and has been used in the management of COVID-
19. Considering the overall health benefits, the herb cannot be claimed to be toxic.[38]

During the ongoing COVID-19 pandemic Tinospora cordifolia also known as Giloy gained
immense popularity and use due to its immunity-boosting function and anti-viral properties. T.
cordifolia is among the most important medicinal plants that has numerous therapeutic

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applications in health due to the production of a diverse array of secondary metabolites.

Therefore, to gain genomic insights into the medicinal properties of T. cordifolia, the first
genome sequencing was carried out using 10x Genomics linked read technology and the draft
genome assembly comprised of 1.01 Gbp. This is also the first genome sequenced from the
plant family Menispermaceae. The deep sequencing of transcriptome from the leaf tissue was
also performed followed by transcriptomic analysis to gain insights into the gene expression
and functions. Further, the phylogenetic position of T. cordifolia was also determined through
the construction of a genome-wide phylogenetic tree using 35 other dicot species and one
monocot species as an outgroup species.[39]

Plants produce a diverse range of bioactive molecules, making them a rich source of different
types of medicines. A regular and widespread use of herbs throughout the world has increased
serious concern over their quality, safety and efficacy. Thus, a proper scientific evidence or
assessment has become the criteria for acceptance of herbal health claims. The anti-oxidant
effects of leaves of Tinospora cordifolia has been examined. [40]

The phytochemical analysis of the plants is very important commercially and has great interest
in pharmaceutical companies for the production of the new drugs for curing of various diseases.
Phytochemicals have two categories i.e., primary and secondary constituents. Primary
constituents have chlorophyll, proteins sugar and amino acids. Secondary constituents contain
terpenoids and alkaloids. Medicinal plants have antifungal, antibacterial and anti-inflammation
activities. Study involves the qualitative phytochemical analysis of two different medicinal
plants: Tinospora cordifolia and Withania somnifera. Data indicates the presence of flavonoids,
alkaloids, proteins, phenolic compounds, cardiac glycosides and tannins. [41]

Tinospora cordifolia is known as Giloe and Guduchi, with significant importance in the
traditional medicinal systems. It is dioeciously plant. It is mostly used in Ayurved system. It is
also known as a ‘Rasayans’ of medicinal system, which develops immune system of the body
and protect against infection. Study is carried out to analyse the phytochemical compounds in
leaves and stem extracts of T. cordifolia by using phytochemical screening tests and estimate
total flavonoid content (TFC) by using aluminium chloride method in the sample extracts. The
leaf and stem extracts of T. cordifolia expressed the presence of several phytochemicals viz.,
flavonoids, amino acids, diterpines, protein, saponins and carbohydrates. The investigation
further proposed that the phytochemicals present in stems and leaves of T. cordifolia, which
can be use as natural antioxidants in medicinal drugs. [42]

The traditional system of medicine in Sri Lanka has shown much better improvement, has
fewer side effects, and is less expensive than modern synthetic drugs in the treatment of many
diseases. The objective of the was to comparatively evaluate the qualitative and quantitative
analysis of phytochemical constituents of leaves of Murraya koenigii (L.) Spreng., Tinospora
cordifolia (Wild) Hook.f., Enicostemma axillare (Lam) A. Raynal, and Gymnema sylvestre R.
Br. were collected from Jaffna District. [43]

Patanjali Giloy Ghanvati

Giloy ghanvati is used as a treatment for general fever and immunity. Useful in generalized
debility, fever, skin & urinary disorders. It is also beneficial in general weakness, fever, dengue,
chicken guinea. It helps build strength and enhance energy, boosts immunity naturally, offers
protection against numerous diseases, alleviates weakness and fatigue. [44]

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Patanjali Ayurveda Giloy Ghanvati is an ayurvedic supplement that helps boost immunity and
protect from various infections. Its key ingredient is Giloy. It helps build strength and stamina
to improve energy levels. The tablets aid in post-illness recovery and manage fever, cough, and
cold. Giloy, an active ingredient in the tablet, has an antipyretic activity that helps to reduce
fever and recover quickly after an infection or illness. It is known to alleviate the symptoms of
the common cold, low immunity, general weakness, and fatigue. This ayurvedic tablet is also
known to increase platelet count and reduce dengue fever. It also aids in toxin removal,
promoting better skin and skin regeneration by increasing collagen production. It can be used
for general weakness and the common cold, Aids in relieving constipation with mucus in the
stool, helps to manage cholesterol levels and keep your heart healthy, it is known to lower your
risk of recurrent infections, boosts your immune system and prevents immune-deficiency
disorders, acts as supportive therapy for individuals with autoimmune disorders, it improves
your appetite and aids in overall growth, plays a key role in the treatment of skin conditions,
helps deal with urinary problems, including urinary tract infections. [45]

The current severe acute respiratory syndrome disease caused by Coronavirus-2 (SARS-CoV-
2) has been a serious strain on the healthcare infrastructure mainly due to the lack of a reliable
treatment option. Alternate therapies aimed at symptomatic relief are currently prescribed
along with artificial ventilation to relieve distress. Traditional medicine in the form of
Ayurveda has been used since ancient times as a holistic treatment option rather than targeted
therapy. The practice of Ayurveda has several potent herbal alternatives for chronic cough,
inflammation, and respiratory distress which are often seen in the SARS-CoV-2 infection. The
aqueous extracts of Tinospora cordifolia (willd.) Hook. f. and Thomson in the form of Giloy
Ghanvati, as a means of treatment to the SARS-CoV-2 spike-protein induced disease
phenotype in a humanized zebrafish model is used. The resultant changes in the disease
phenotype were comparable to the group that were given the reference compound,
Dexamethasone. These findings correlated well with various phyto-compounds detected in the
Giloy Ghanvati and their reported roles in the viral disease phenotype amelioration.[46]

Present study deals with Colorimetric phytochemical analysis of Tinospora Cordifolia (Giloy
leaf extract) and FTIR study of its medicinal sample Giloy Ghanvati. Colorimetric
phytochemical analysis of Tinospora Cordifolia (Giloy leaf extract) involves preparation of
standard Giloy Ghanvati solution, Ferric Chloride solution and Giloy leaf Solution. Different
systems were prepared. Absorbance of standard Giloy solution was taken at different
wavelength. And λmax was calculated. Which was 420 nm. Absorbance of standard Giloy
solution of different concentrations were taken at 420 nm and Calibration plot was prepared.
Then absorbance of Giloy leaf solutions were taken at 420 nm and concentration of Giloy in
Giloy leaf was calculated from calibration plot. Tinospora cordifolia which contains diverse
phytochemicals, including alkaloids, phytosterols, glycosides, tinosporide, and other mixed
chemical compounds reduces ferric ions and production of violet coloration by addition of
ferric chloride to dilute solution of Giloy takes place and the concentration of Giloy was
investigated colorimetrically. The method is simple, rapid and precise. Interpretation of FTIR
Spectra of Giloy Ghanvati medicinal sample shows presence of various functional groups such
as O-H stretching - Alcohol, Carboxylic acid, O-H bending - Alcohol, N-H stretching - Amine
salt, C=C stretching - α, β- unsaturated ketone and C=O stretching - Conjugated acid.

METHODOLOGY

1. Colorimetric phytochemical analysis of Tinospora Cordifolia (Giloy)

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Colorimetric phytochemical analysis of Tinospora Cordifolia (Giloy leaf extract) involves

preparation of standard Giloy Ghanvati solution, Ferric Chloride solution and Giloy leaf
Solution. Different systems were prepared. Absorbance of standard Giloy solution was taken
at different wavelength. And λmax was calculated. Which was 420 nm. Absorbance of standard
Giloy solution of different concentrations were taken at 420 nm and Calibration plot was
prepared. Then absorbance of Giloy leaf solutions were taken at 420 nm and concentration of
Giloy in Giloy leaf was calculated from calibration plot. Tinospora cordifolia which contains
diverse phytochemicals, including alkaloids, phytosterols, glycosides, tinosporide, and other
mixed chemical compounds reduces ferric ions and production of violet coloration by addition
of ferric chloride to dilute solution of Giloy takes place and the concentration of Giloy was
investigated colorimetrically. The method is simple, rapid and precise.

Preparation of Standard Giloy Ghanvati solution: Weight of 1 tablet of Giloy Ghanvati was
taken and crushed it then 0.1 g powder was dissolved in 100 ml distilled water.
Preparation of Ferric Chloride solution: 1 g Ferric chloride was dissolved in 20 ml distilled
water.
Preparation of Giloy Leaf Solution: 2 Giloy leaf was crushed with the help of distilled water,
transferred completely in 100 ml volumetric flask and volume was made up to 100 ml with the
help of distilled water.

Following systems were prepared

System Standard Giloy Giloy Leaf Ferric Chloride
Distilled water
No. Ghanwati Solution, ml Solution solution, ml
1 1 - 1 8
2 2 - 1 7
3 3 - 1 6
4 4 - 1 5
5 5 - 1 4
6 6 - 1 3
7 - 1 1 8
8 - 2 1 7
9 - 3 1 6
10 - 4 1 5
11 - 5 1 4
12 - 6 1 3

Absorbance of system No 1 taken at different wave length using water as blank. λmax was
noted which is 420 nm. Absorbance of system 1 to 6 was taken at λmax 420 nm. Then
absorbance of system 7 to 12 was taken at 420 nm wavelength. Calibration plot was prepared
between Concentration of Giloy and Absorbance. And from this calibration plot concentration
of Giloy from Giloy Leaf sample was calculated.

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Absorbance of System Absorbance of System

Wave length, nm Wave length, nm
No. 1 No. 1
400 0.44 530 0.48
420 0.62 620 0.28
470 0.59 660 0.16
500 0.54 700 0.08

λmax 420 nm
0.7

0.6

0.5
Absorbance

0.4

0.3

0.2

0.1

0
0 100 200 300 400 500 600 700 800
Wavelength, nm

System No. Absorbance at λmax 420 nm System No. Absorbance at λmax 420 nm
1 0.72 7 0.68
2 0.77 8 0.88
3 0.79 9 1.04
4 0.81 10 1.13
5 0.84 11 1.24
6 0.86 12 1.27

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2. FTIR study of Giloy Ghanvati medicinal sample

FTIR can be routinely used to identify the functional groups and identification/quality control
of raw material / finished products. Spectrum RX-I offers fast throughput and rapid access to
reliable and dependable IR results. High signal to noise ratio makes FTIR more useful for
difficult samples. It has resolution of 1 cm1 and scan range of 4000 cm-1 to 250 cm-1. In the
normal mode around 10 mg sample is required in the form of fine powder. The sample can be
analyzed in the form of liquid, solid and thin films also.

FTIR spectra of Giloy Ghanvati medicinal sample is obtained at room temperature by using an
FTIR Spectrophotometer – Perkin Elmer – Spectrum RX-IFTIR. The spectra is collected in a
range from 400 to 4000 cm-1.

FTIR spectra of Giloy Ghanvati medicinal sample

RESULTS AND DISCUSSION

1. Colorimetric phytochemical analysis of Tinospora Cordifolia (Giloy)

1 Tablet of Giloy Ghanvati contains 0.5 g Giloy (Quoted value).
Weight of 1 tablet of Giloy Ghanvati was 0.6 g. Hence 0.6 g tablet contains 0.5 g Giloy
0.1 g tablet powder will contains 0.1 x 0.5 / 0.6 = 0.0833 g Giloy.

System Volume of standard Giloy Absorbance at

Weight of Giloy, g
No Ghanvati solution taken, ml 420 nm
1 1 1 x 0.0833 / 100 = 0.000833 0.75
2 2 2 x 0.0833 / 100 = 0.001666 0.77
3 3 3 x 0.0833 / 100 = 0.002499 0.79
4 4 4 x 0.0833 / 100 = 0.003332 0.81
5 5 5 x 0.0833 / 100 = 0.004165 0.84
6 6 6 x 0.0833 / 100 = 0.004998 0.86

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Calibration Plot
0.88

0.86

0.84

0.82
Absorbance

0.8

0.78

0.76

0.74

0.72
0 0.001 0.002 0.003 0.004 0.005 0.006
Concentration of Giloy, g

Calibration Plot
1.4

1.2

1
Absorbance

0.8

0.6

0.4

0.2

0
0 0.005 0.01 0.015 0.02 0.025
Concentration of Giloy, g

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Volume of Average
Absorb Concentration
Syste Giloy leaf Concentration of Giloy concentration
ance at of Giloy from
m No solution per100 ml, g of Giloy per
420 nm graph, g
taken, ml 100 ml, g
7 0.68 - 1 -
8 0.88 0.0056 2 100 x 0.0056 / 2 = 0.28
9 1.04 0.0114 3 100 x 0.0114 / 3 = 0.38
10 1.13 0.0145 4 100 x 0.0145 / 4 = 0.3625 0.3435
11 1.24 0.0185 5 100 x 0.0185 / 5 = 0.37
12 1.27 0.0195 6 100 x 0.0195 / 6 = 0.325
Hence 100 ml Giloy leaf solution contains 0.3435 g Giloy. Hence 2 Giloy leaf contains 0.3435
g Giloy. Hence 1 Giloy leaf will contain 0.3435 / 2 = 0.1718 g Giloy i.e., diverse
phytochemicals, including alkaloids, phytosterols, glycosides, tinosporide, and other mixed
chemical compounds.

2. FTIR study of Giloy Ghanvati medicinal sample

Spectral
Pattern and intensity of Bond causing
region wave Compound Class
Band Absorption
number cm-1
3940.57 Broad and low intensity - -
3900.07 Broad and low intensity - -
3857.63 Broad and low intensity - -
3836.42 Broad and low intensity - -
3741.90 Broad and low intensity - -
3369.64 Broad and strong intensity O-H stretching Alcohol
O-H stretching Carboxylic acid
2926.01 Broad and strong intensity
N-H stretching Amine salt
2360.87 Sharp and moderate intensity - -
2266.36 - - -
2140.99 Broad and low intensity - -
1950.03 - - -
1915.31 - - -
1703.14 Broad and strong intensity C=O stretching Conjugated acid
1612.49 Broad and strong intensity C=C stretching α, β- unsaturated ketone

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1512.19 Broad and low intensity - -

1419.61 Broad and moderate intensity O-H bending alcohol
1367.53 Broad and moderate intensity - -
1203.58 - - -
1145.72 Broad and moderate intensity - -
1074.35 Broad and moderate intensity - -
1033.85 Broad and moderate intensity - -
929.69 Broad and low intensity - -
856.39 Broad and low intensity - -
763.81 Broad and low intensity - -
663.51 Broad and low intensity - -
569.00 Broad and low intensity - -
526.57 Broad and low intensity - -
460.99 Broad and low intensity - -
435.91 Broad and low intensity - -

CONCLUSION

1. Colorimetric phytochemical analysis of Tinospora Cordifolia (Giloy)

1 Giloy leaf contains 0.1718 g Giloy i.e., diverse phytochemicals, including alkaloids,
phytosterols, glycosides, tinosporide, and other mixed chemical compounds.

2. FTIR study of Giloy Ghanvati medicinal sample

Interpretation of FTIR Spectra of Giloy Ghanvati medicinal sample shows presence of various
functional groups such as O-H stretching - Alcohol, Carboxylic acid, O-H bending - Alcohol,
N-H stretching - Amine salt, C=C stretching - α, β- unsaturated ketone and C=O stretching -
Conjugated acid.

REFERENCES

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2. ^ Mishra, Ashish; Gond, Surendra K.; Kumar, Anuj; Sharma, Vijay K.; Verma, Satish K.; Kharwar,
Ravindra N.; Sieber, Thomas N. (2012). "Season and Tissue Type Affect Fungal Endophyte
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4. ^ Swaminathan, K.; Sinha, U. C.; Bhatt, R. K.; Sabata, B. K.; Tavale, S. S. (1989). "Structure of
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5. ^ Kumar, Pradeep; Kamle, Madhu; Mahato, Dipendra K.; Bora, Himashree; Sharma, Bharti;
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Ethnopharmacology, Clinical Application and Conservation Strategies". Current Pharmaceutical
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6. ^ Jump up to:a b Banjot Kaur (17 February 2022). "As COVID Surged, India Had a Silent Outbreak
of Giloy-Induced Liver Injury". Science: The Wire.
7. ^ Nagral, Aabha; Adhyaru, Kunal; Rudra, Omkar S.; et al. (2021-07-02). "Herbal Immune
Booster-Induced Liver Injury in the COVID-19 Pandemic - A Case Series". Journal of Clinical and
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6883. PMC 8252698. PMID 34230786.
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says study". Deccan Herald. The Printers, Mysore.
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cordifolia ( giloy)–induced liver injury during the COVID‐19 pandemic — Multicenter nationwide
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1300. doi:10.1002/hep4.1904. PMC 9134809. PMID 35037744.
10. The chemical constituents and diverse pharmacological importance of Tinospora cordifolia,
Priyanka Sharma, Bharat P. Dwivedee, Dheeraj Bisht, Ashutosh K. Dasha, Deepak Kumar, Heliyon
· September 2019. DOI: 10.1016/j.heliyon.2019.e02437
11. M.Q.I. Khuda, A. Khaleque, N. Ray, Tinospora cordifolia constituents of plants fresh from the field,
Sci. Res. 1 (1964) 177–183.
12. J.B. Hanuman, R.K. Bhatt, B.K. Sabata, A diterpenoid furanolactone from Tinospora cordifolia,
Phytochemistry 25 (1986) 1677–1680.
13. R.K. Bhatt, J.B. Hanuman, B.K. Sabata, A new clerodane derivative from Tinospora cordifolia,
Phytochemistry 27 (1988) 1212–1216.
14. J.B. Hanuman, R.K. Bhatt, B. Sabata, A clerodane furano-diterpene from Tinospora cordifolia, J.
Nat. Prod. 51 (1988) 197–201.
15. R.K. Bhatt, B.K. Sabata, A furanoid diterpene glucoside from Tinospora cordifolia, Phytochemistry
28 (1989) 2419–2422.
16. M.A. Khan, I.A. Gray, P.G. Waterman, Tinosporaside an 18-norclerodane glucoside from
Tinospora cordifolia, Phytochemistry 28 (1989) 273–275.
17. V.D. Gangan, P.P. Arjun, T. Sipahimalani, A. Banerji, A. Cardifolisides, C. B, Norditerpene furon
glucoside from Tinospora cordifolia, Phytochemistry 37 (1994) 781–786.
18. R. Maurya, S.S. Handa, Tinocordifolin, a sesquiterpene from Tinospora cordifolia, Phytochemistry
44 (1998) 1343–1345.
19. V.D. Gangan, P.P. Arjun, A.T. Sipahimalani, A. Banerji, Norditerpene furon glucoside from
Tinospora cordifolia, Phytochemistry 39 (1995) 1139–1142.
20. V. Wazir, R. Maurya, R.S. Kapil, A clerodane furano diterpene glucoside from Tinospora
cordifolia, Phytochemistry 38 (1995) 447–449.
21. V.D. Gagan, P. Pradhan, A.T. Sipahimalan, A. Banerji, F. Palmatosides C, Diterpene furan
glucosides from Tinospora cordifolia-structural elucidation by 2D NMR spectroscopy, Indian J.
Chem. 35B (1996) 630–634.
22. N. Choudhary, M.B. Siddiqui, S. Azmat, S. Khatoon, Tinospora cordifolia: ethnobotany,
phytopharmacology and phytochemistry aspects, IJPSR 4 (2013) 891.
23. N.G. Bisset, J. Nwaiwu, Quaternary alkaloids of Tinospora species, Planta Med. 48 (1983) 275–
279.
24. V.R. Mahajan, C.I. Jolly, K.M. Kundnani, A new hypoglycaemic agent from Tinospora cordifolia,
Indian Drugs 23 (1985) 119–120.
25. D.N.K. Sarma, R.L. Khosa, J.P.N. Chansauria, A.K. Ray, The effect of Tinospora cordifolia on
brain neurotransmitters in the stressed rat, Fitoterapia 66 (1995) 421–422.

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26. A.K. Pathak, A.K. Agarwal, D.C. Jain, R.P. Sharma, O.W. Howarth, NMR studies of 20
hydroxyecdysones, a steroid isolated from Tinospora cordifolia, Indian J. Chem. 34 (1995) 674–
676.
27. J.B. Hanuman, A.K. Mishra, B. Sabata, A natural phenolic lignan from Tinospora cordifolia Miers,
J. Chem. Soc. (1986) 1181–1185.
28. A.R. Kidwai, K.C. Salooja, V.N. Sharma, S. Siddiqui, Chemical examination of Tinospora
cordifolia, J. Sci. Indian Res. 8 (1949) 115–118.
29. A. Khaleque, M.A.W. Maith, M.S. Huq, B. K Abul, Tinospora cordifolia IV. Isolation of
heptacosanol, ß sitosterol and three other compounds tinosporine, cordifol and cordifolone,
Pakistan J. Sci.Industry Res. 14 (1970) 481–483.
30. S.N. Dixit, R.L. Khosa, Chemical investigations on Tinospora cordifolia (wild.) miers, Indian. J.
Appl. Chem. 34 (1971) 46–47.
31. A.K. Pathak, D.C. Jain, P.R. Sharma, Chemistry and biological activities of the genus Tinospora,
Int. J. Pharmacogn. 33 (1995) 277–287.
32. M.Q. Khuda, A. Khaleque, K.A. Basar, M.A. Rouf, M.A. Khan, N. Roy, Studies on Tinospora
cordifolia II: isolation of tinosporine, tinosporic acid and tinosporol from the fresh creeper, Sci.
Res. 3 (1966) 9–12.
33. A. Khaleque, M.A.W. Maith, M.S. Huq, K.A. Tinospora cordifolia III, Isolation of tinosporine,
heptacosanol, ß sitosterol, Pakistan J . Sci. Industry Res. 14 (1971) 481–483.
34. R. Maurya, V. Wazir, A. Tyagi, R.S. Kapil, Clerodane diterpene from Tinospora cordifolia,
Phytochemistry 38 (1995) 659–661.
35. P. Pradhan, V.D. Gangan, A.T. Sipahimalani, A. Banerji, Two phytoecdysones from Tinospora
cordifolia: structural assignment by 2D NMR spectroscopy, Indian J. Chem. 36B (1997) 958–962.
36. G. Chintalwar, A. Jain, A. Sipahimalani, A. Banerji, P. Sumariwalla, R. Ramakrishnan, K. Sainis,
An Immunologically active arabinogalactan from Tinospora cordifolia, Phytochemistry 52 (1999)
1089–1093
37. Antibacterial and qualitative phytochemical analysis of Giloy extract for application of herbal finish
on cotton fabric, K Medhaa, N Aryaa, K Malikb, S Ahlawatc & N Chauhana, Indian Journal of
Traditional Knowledge, Vol 20(4), October 2021, pp 944-950
38. https://pib.gov.in/PressReleasePage.aspx?PRID=1798676
39. Genome sequencing and assembly of Tinospora cordifolia (Giloy) plant,
Shruti Mahajan, Abhisek Chakraborty, Titas Sil, Vineet K Sharma,
https://www.biorxiv.org/content/10.1101/2021.08.02.454741v1
40. Studies on Anti-oxidant activity of Tinospora cordifolia (Miers.) Leaves using in vitro models,
Ramya Premanath and N. Lakshmidevi, Journal of American Science, 2010;6(10):736-743.
41. Qualitative phytochemical analysis of Tinospora cordifolia and Withania somnifera, Iqra Nazir and
Rikhi S Chauhan, The Pharma Innovation Journal 2018; 7(10): 333-336
42. Qualitative and quantitative analysis of leaves and stem of Tinospora Cordifolia in different solvent
extract, Garg Praveen, Garg Rajesh, Journal of Drug Delivery & Therapeutics. 2018; 8(5-s):259-
264
43. Comparative Analysis of Qualitative and Quantitative Phytochemical Evaluation of Selected
Leaves of Medicinal Plants in Jaffna, Sri Lanka, Gowri Rajkumar, Panambara Arachchilage Harini,
Rangana Panambara, Vinotha Sanmugarajah, Borneo Journal of Pharmacy, Vol 5 Issue 2 May
2022, Page 93 – 103
44. https://www.patanjaliayurved.net/product/ayurvedic-medicine/vati/patanjali-giloy-ghanvati-60-
n/624
45. https://www.1mg.com/otc/patanjali-ayurveda-giloy-ghanvati-
otc324816?wpsrc=Google+Organic+Search
46. Giloy Ghanvati (Tinospora cordifolia (Willd.) Hook. f. and Thomson) Reversed SARS-CoV-2 Viral
Spike-Protein Induced Disease Phenotype in the Xenotransplant Model of Humanized Zebrafish,
Acharya Balkrishna, Lakshmipathi Khandrika, and Anurag Varshney, Front Pharmacol, 2021; 12:
635510.

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61

Determination of Hydochloric Acid Neutralizing Capacity of Different

Antacid Tablet by Back Titration Method and Its FTIR Analysis

Dr Swaroopa Rani N. Gupta

Professor, Department of Chemistry, Brijlal Biyani Science College, Amravati, Maharashtra India
[email protected]

ABSTRACT
An antacid is a substance which neutralizes stomach acidity and is used to relieve heartburn,
indigestion or an upset stomach. Several liquid antacid preparations are marketed. Common
liquid preparations include milk of magnesia and magnesium/aluminum combinations. A
potential advantage of using a liquid preparation over a tablet is that liquids may provide
quicker relief, however this may coincide with a shorter duration of action. Chewable tablets
are one of the most common forms of antacids, and are readily available over the counter. Upon
reaching the stomach, the tablet powder will dissolve in the stomach acid, allowing the cations
to be released and neutralize excess stomach acid. Common salts available in tablet form
include those of calcium, magnesium, aluminum, and sodium. Effervescent tablets are tablets
which are designed to dissolve in water, and then release carbon dioxide. Common ingredients
include citric acid and sodium bicarbonate, which react when in contact with water to produce
carbon dioxide. Effervescent antacids may also contain aspirin, sodium carbonate, or tartaric
acid. Those containing aspirin may cause further gastric irritation and ulceration due to aspirin's
effects on the mucous membrane of the stomach.
Present study deals with determination of Hydrochloric acid neutralizing capacity of an antacid
tablets such as ACILOC 150, Athzol-DSR, Gelusil, NEXPRO-40, Omee-D, PAN-L, Pantafol-
DSR, Rabesec-20, Rabifeel – LS and Rantac 150 using back titration method and FTIR study
of Pantafol-DSR.
Antacid tablets contain bases like calcium carbonate, magnesium carbonate, magnesium
hydroxide, sodium bicarbonate etc. The tablet can neutralize the HCl present in stomach and
can decreases the acidity. The HCl neutralizing capacity of an antacid tablet can be determined
by adding tablet to a known quantity of excess HCl then back titrating the unneutralized acid
with standard alkali solution using phenolphthalein indicator. The higher the amount of
Hydrocloric acid neutralized by Antacids, the better is the acid neutralizing capacity. The result
shows that the tablet Rantac 150 has highest acid neutralizing capacity and PAN_L lowest acid
neutralizing capacity. Rantac 150 >Rabifeel – LS > Omee-D > Pantafol-DSR > Rabesec-20 >
NEXPRO-40 > Athzol-DSR > ACILOC 150 > Gelusil > PAN-L. The back titration method
used in this study is simple, inexpensive and can be used in routine monitoring of the quality
of Antacid tablets.
FTIR spectra of Pantafol-DSR is obtained at room temperature by using an FTIR
Spectrophotometer – Perkin Elmer – Spectrum RX-IFTIR. The spectra is collected in a range
from 400 to 4000 cm-1. Interpretation of FTIR Spectra of Pantafol-DSR shows Presence of
various functional groups such as C-H bending - Aromatic compound; O-H stretching –
Alcohol, Carboxylic acid, C=O stretching - Conjugated acid, Conjugated aldehyde, C≡C
stretching – Alkyne.

Keywords: Hydochloric Acid Neutralizing Capacity, Antacid Tablet, Back Titration

Method, FTIR Analysis

INTRODUCTION

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Antacid
An antacid is a substance which neutralizes stomach acidity and is used to relieve heartburn,
indigestion or an upset stomach.[1] Some antacids have been used in the treatment of
constipation and diarrhea.[2] Marketed antacids contain salts of aluminium, calcium,
magnesium, or sodium.[2] Some preparations contain a combination of two salts, such as
magnesium carbonate and aluminium hydroxide (e. g. hydrotalcite).[3] Antacids are available
over the counter and are taken by mouth to quickly relieve occasional heartburn, the major
symptom of gastroesophageal reflux disease and indigestion. Treatment with antacids alone is
symptomatic and only justified for minor symptoms.[4] Alternative uses for antacids include
constipation, diarrhea, hyperphosphatemia, and urinary alkalization.[5] Some antacids are also
used as an adjunct to pancreatic enzyme replacement therapy in the treatment of pancreatic
insufficiency.[6] Non-particulate antacids (sodium citrate) increase gastric pH with little or no
effect on gastric volume, and therefore may see some limited use in pre-operative procedures.
Sodium citrate should be given within 1 hour of surgery to be the most effective.[7]
Conventional effervescent tablets contain a significant amount of sodium and are associated
with increased odds of adverse cardiovascular events according to a 2013 study.[8] Alternative
sodium-free formulations containing magnesium salts may cause diarrhea, whereas those
containing calcium or aluminum may cause constipation. Rarely, long-term use of calcium
carbonate may cause kidney stones. Long-term use of antacids containing aluminum may
increase the risk of developing osteoporosis.[9] In vitro studies have found a potential for acid
rebound to occur due to antacid overuse, however the significance of this finding has been
called into question.[10][11] When an excess amount of acid is produced in the stomach, the
natural mucous barrier that protects the lining of the stomach can degrade, leading to pain and
irritation. There is also potential for the development of acid reflux, which can cause pain and
damage to the esophagus. Antacids contain alkaline ions that chemically neutralize stomach
gastric acid, reducing damage to the stomach lining and esophagus, and relieving pain.[1] Some
antacids also inhibit pepsin, an enzyme that can damage the esophagus in acid reflux.[5][12]
Antacids do not directly inhibit acid secretion, and thus are distinct from acid-reducing drugs
like H2-receptor antagonists or proton pump inhibitors. Antacids do not kill the bacteria
Helicobacter pylori, which causes most ulcers.[4] Antacids are known to interact with several
oral medications, including fluoroquinolone and tetracycline antibiotics, iron, itraconazole, and
prednisone.[13] Metal chelation is responsible for some of these interactions
(e.g.fluoroquinolones, tetracyclines), leading to decreased absorption of the chelated drug.
Some interactions may be due to the pH increase observed in the stomach following antacid
ingestion, leading to increased absorption of weak acids, and decreased absorption of weak
bases. Antacids also cause an increase in pH of the urine (alkalization), which may cause
increased blood concentrations of weak bases, and increased excretion of weak acids.[14] A
proposed method to mitigate the effects of stomach acidity and chelation on drug absorption is
to space out the administration of antacids with interacting medications, however this method
has not been well studied for drugs affected by urine alkalization.[13] There are concerns
regarding interactions between delayed-release tablets and antacids, as antacids may increase
the stomach pH to a point at which the coating of the delayed-release tablet will dissolve,
leading to degradation of the drug if it is pH sensitive.[14] Antacids may be formulated with
other active ingredients such as simethicone to control gas, or alginic acid to act as a physical
barrier to acid.[15]

Several liquid antacid preparations are marketed. Common liquid preparations include milk of
magnesia and magnesium/aluminum combinations. A potential advantage of using a liquid
preparation over a tablet is that liquids may provide quicker relief, however this may coincide

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with a shorter duration of action.[16] Chewable tablets are one of the most common forms of
antacids, and are readily available over the counter. Upon reaching the stomach, the tablet
powder will dissolve in the stomach acid, allowing the cations to be released and neutralize
excess stomach acid. Common salts available in tablet form include those of calcium,
magnesium, aluminum, and sodium.[13] Some common brand are Tums, Gaviscon chewable
tablets, and Maalox chewable tablets.[17] Effervescent tablets are tablets which are designed
to dissolve in water, and then release carbon dioxide.[18][19][20] Common ingredients include
citric acid and sodium bicarbonate, which react when in contact with water to produce carbon
dioxide. Effervescent antacids may also contain aspirin,[21] sodium carbonate, or tartaric
acid.[22] Those containing aspirin may cause further gastric irritation and ulceration due to
aspirin's effects on the mucous membrane of the stomach.[23] Common brands include Alka-
Seltzer, Gaviscon, and Eno.

Study based on the evaluation of acid neutralizing capacity of five different commercial brands
of antacid tablets was done. [24] Antacids are commonly used as over - the - counter (OTC)
drugs or prescribed medications. Some antacid products may neutralize more acid in the
stomach than others. The ability of an antacid to neutralize acid is expressed as its Acid
Neutralizing Capacity (ANC). Study was undertaken with the objective of assessing the quality
of different brands of antacid tablets. The assessment parameters included the evaluation of
uniformity of weight, uniformity of thickness, crushing strength, friability, as well as the ANC
(using pH and titrimetric method), which is easy to use, accurate, reproducible, simple, and
inexpensive. [25] Study has been evaluated the acid neutralizing and buffering capacities (BC)
of nine selected antacid brands (“L1-L9”) in Ghana. [26] Gastric acidity is highly prevalent in
the Indian Population. People usually take commercially available antacids to alleviate the
symptoms. Unfortunately, the use of these over the counter drugs has been so common that
they are vastly overused and are taken even for mild heartburn or indigestion, which makes the
underlying problem worse. The study aimed at identifying common traditional remedies used
in India to treat acidity and to evaluate their acid neutralizing capacity; and compare it with
their commercially available counterparts. The acid neutralizing capacities of the traditional
remedies were analyzed by Back titration method. [27] The study was aimed to evaluate the
acid-neutralizing capacity (ANC) and other properties of antacid drugs marketed in Morocco.
Other properties such as price and sodium content were also studied. [28]

Present study deals with determination of Hydrochloric acid neutralizing capacity of an antacid
tablets such as ACILOC 150, Athzol-DSR, Gelusil, NEXPRO-40, Omee-D, PAN-L, Pantafol-
DSR, Rabesec-20, Rabifeel – LS and Rantac 150 using back titration method and FTIR study
of Pantafol-DSR. Antacid tablets contain bases like calcium carbonate, magnesium carbonate,
magnesium hydroxide, sodium bicarbonate etc. The tablet can neutralize the HCl present in
stomach and can decreases the acidity. The HCl neutralizing capacity of an antacid tablet can
be determined by adding tablet to a known quantity of excess HCl then back titrating the
unneutralized acid with standard alkali solution using phenolphthalein indicator. The higher
the amount of Hydrocloric acid neutralized by Antacids, the better is the acid neutralizing
capacity. The result shows that the tablet Rantac 150 has highest acid neutralizing capacity and
PAN_L lowest acid neutralizing capacity value. Rantac 150 >Rabifeel – LS > Omee-D >
Pantafol-DSR > Rabesec-20 > NEXPRO-40 > Athzol-DSR > ACILOC 150 > Gelusil > PAN-
L. The back titration method used in this study is simple, inexpensive and can be used in routine
monitoring of the quality of Antacid tablets. FTIR spectra of Pantafol-DSR is obtained at room
temperature by using an FTIR Spectrophotometer – Perkin Elmer – Spectrum RX-IFTIR. The
spectra is collected in a range from 400 to 4000 cm-1. Interpretation of FTIR Spectra of
Pantafol-DSR shows Presence of various functional groups such as C-H bending - Aromatic

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compound; O-H stretching – Alcohol, Carboxylic acid, C=O stretching - Conjugated acid,
Conjugated aldehyde, C≡C stretching – Alkyne.

METHODOLOGY

1. Determination of Hydrochloric acid neutralizing capacity of an antacid tablet

Antacid tablets contain bases like calcium carbonate, magnesium carbonate, magnesium
hydroxide, sodium bicarbonate etc. The tablet can neutralize the HCl present in stomach and
can decreases the acidity. The HCl neutralizing capacity of an antacid tablet can be determined
by adding tablet to a known quantity of excess HCl then back titrating the unneutralized acid
with standard alkali solution using phenolphthalein indicator.

CaCO3 + 2HCl → CaCl2 + H2O + CO2(g)

MgCO3 + 2HCl → MgCl2 + H2O + CO2
Mg(OH)2 + 2HCl → MgCl2 + 2H2O
NaHCO3 + HCl → NaCl + H2O + CO2

The above reactions are common reactions between an antacid in which main ingredients are
CaCO3, MgCO3, Mg(OH)2, NaHCO3 and stomach HCl

Excess HCl + NaOH → NaCl + H2O

Preparation of 0.1 M Oxalic acid solution: 0.1 M Oxalic acid solution was prepared by
dissolving 0.63 g oxalic acid with distilled water in 100 ml volumetric flask and volume was
made upto 100 ml mark with distilled water.

Preparation of 0.1 M Sodium hydroxide solution: 0.1 M Sodium hydroxide solution was
prepared by dissolving 4 g sodium hydroxide with distilled water in 1 litre volumetric flask and
volume was made upto 1litre mark with distilled water.

Preparation of 0.1 M Hydrochloric acid solution: 0.1 M Hydrochloric acid solution was
prepared by diluting 8.3 ml of 12 M Hydrochloric acid with distilled water in 1 litre volumetric
flask and volume was made upto 1litre mark with distilled water.

Standardization of Sodium hydroxide solution: 10 ml of 0.1 M Oxalic acid solution was

taken in 100 ml conical flask to it 3 drops of phenolphthalein indicator was added and solution
was titrated with Sodium hydroxide solution until it turns pink which persisted for at least 30
seconds. The volume of Sodium hydroxide solution was recorded. The titration procedure was
repeated 2 more times and the constant titre value was recorded.

Standardization of Hydrochloric acid solution: 10 ml of Hydrochloric acid solution was

taken in 100 ml conical flask to it 3 drops of phenolphthalein indicator was added and solution
was titrated with Sodium hydroxide solution until it turns pink which persisted for at least 30
seconds. The volume of Sodium hydroxide solution was recorded. The titration procedure was
repeated 2 more times and the constant titre value was recorded.

Determination of Hydrochloric acid neutralizing capacity of an antacid tablet: One

Antacid tablet of Pantafol-DSR, Omee-D, Rabifeel – LS, Rantac 150, Gelusil, Athzol-DSR,
PAN-L, Rabesec-20, ACILOC 150 and NEXPRO-40 were crushed in mortal and pestle. 0.5 g
crushed tablet was transferred into 250 ml conical flask. To it 75 ml 0.1 N HCl was added and

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solution was heated to boil and dissolve the crushed tablet for 5 minutes then cooled and to it
3 drops of phenolphthalein indicator was added and solution was titrated with Sodium
hydroxide solution until it turns pink which persisted for at least 30 seconds. The volume of
Sodium hydroxide solution was recorded. The titration procedure was repeated 2 more times
and the average titre value was recorded. Molarity of HCl in reaction mixture after
neutralization with Antacid tablet was calculated. From which difference in Molarity of HCl,
weight of HCl neutralized by Antacid Tablet per litre, weight of HCl neutralized by Antacid
Tablet per 75 ml and weight of HCl neutralized by 1 g Antacid tablet was calculated.

2. FTIR study of Pantafol-DSR

FTIR can be routinely used to identify the functional groups and identification/quality control
of raw material/finished products. Spectrum RX-I offers fast throughput and rapid access to
reliable and dependable IR results. High signal to noise ratio makes FTIR more useful for
difficult samples. It has resolution of 1 cm1 and scan range of 4000 cm-1 to 250 cm-1. In the
normal mode around 10 mg sample is required in the form of fine powder. The sample can be
analyzed in the form of liquid, solid and thin films also.

FTIR spectra of Pantafol-DSR is obtained at room temperature by using an FTIR

Spectrophotometer – Perkin Elmer – Spectrum RX-IFTIR. The spectra is collected in a range
from 400 to 4000 cm-1.

Figure 1. FTIR spectra of Pantafol-DSR

RESULTS AND DISCUSSION

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1. Determination of Hydrochloric acid neutralizing capacity of an antacid tablet

Standardization of Sodium hydroxide solution:
Volume of 0.1 M Oxalic Volume of Sodium hydroxide
Constant titre value
acid solution taken, ml solution required, ml
10 10.0
10 10.1 10.1
10 10.1

Standardization of Hydrochloric acid solution:

Volume of Hydrochloric Volume of Sodium hydroxide
Constant titre value
acid solution taken, ml solution required, ml
10 10.0
10 9.9 9.9
10 9.9

Determination of Hydrochloric acid neutralizing capacity of an antacid tablet:

Molarity of Oxalic acid = 0.63 x 10 / 63 = 0.1
Molarity of Sodium hydroxide solution = 0.1 x 10 / 10.1 = 0.099
Molarity of Hydrochloric acid solution = 0.099 x 9.9 / 10 = 0.098

Table 1. Determination of Hydrochloric acid neutralizing capacity of an antacid tablet

Constant Molarity of HCl Weight of
titre in reaction HCl Weight of HCl Weight of HCl
Name of Difference in
value, mixture after neutralized neutralized by neutralized by 1
Antacid Molarity of
ml neutralization by Antacid Antacid Tablet g Antacid
Tablet HCl
with Antacid Tablet per 75 ml, g tablet, g
tablet perliter, g

ACILOC 12 0.099 x 12 / 75 0.098 – 0.0158 0.0822 x 36.5 3.0003 x75 / 0.2250 x 1 / 0.5
150 = 0.0158 = 0.0822 = 3.0003 1000 = 0.2250 = 0.45

Athzol- 10.5 0.099 x 10.5 / 0.098 – 0.0139 0.0841 x 36.5 3.0697 x75 / 0.2302 x 1 / 0.5
DSR 75 = 0.0139 = 0.0841 = 3.0697 1000 = 0.2302 = 0.4604

14.5 0.099 x 14.5 / 0.098 – 0.0191 0.0789 x 36.5 2.8799 x75 / 0.2160 x 1 / 0.5
Gelusil
75 = 0.0191 = 0.0789 = 2.8799 1000 = 0.2160 = 0.432

NEXPRO- 8.6 0.099 x 8.6 / 75 0.098 – 0.0114 0.0866 x 36.5 3.1609 x75 / 0.2371 x 1 / 0.5
40 = 0.0114 = 0.0866 = 3.1609 1000 = 0.2371 = 0.4742

2.1 0.099 x 2.1 / 75 0.098 – 0.0028 0.0952 x 36.5 3.4748 x75 / 0.2606 x 1 / 0.5
Omee-D
= 0.0028 = 0.0952 = 3.4748 1000 = 0.2606 = 0.5212

16 0.099 x 16 / 75 0.098 – 0.0211 0.0769 x 36.5 2.8069 x75 / 0.2105 x 1 / 0.5

PAN-L
= 0.0211 = 0.0769 = 2.8069 1000 = 0.2105 = 0.421

Pantafol- 4.8 0.099 x 4.8 / 75 0.098 – 0.0063 0.0917 x 36.5 3.3471 x75 / 0.2510 x 1 / 0.5
DSR = 0.0063 = 0.0917 = 3.3471 1000 = 0.2510 = 0.502

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Rabesec- 6.5 0.099 x 6.5 / 75 0.098 – 0.0086 0.0894 x 36.5 3.2631 x75 / 0.2447 x 1 / 0.5
20 = 0.0086 = 0.0894 = 3.2631 1000 = 0.2447 = 0.4894

Rabifeel - 1 0.099 x 1 / 75 = 0.098 – 0.0013 0.0967 x 36.5 3.5296 x75 / 0.2647 x 1 / 0.5
LS 0.0013 = 0.0967 = 3.5296 1000 = 0.2647 = 0.5294

Rantac 0.5 0.099 x 0.5 / 75 0.098 – 0.0007 0.0973 x 36.5 3.5515 x75 / 0.2664 x 1 / 0.5
150 = 0.0007 = 0.0973 = 3.5515 1000 = 0.2664 = 0.5328

Hydrochloric acid neutralizing capacity of an antacid tablet

0.6
HCl neutralized by 1 g Antacid tablet, g

0.5

0.4

0.3

0.2

0.1

Name of Antacid Tablet

Figure 2 Hydrochloric acid neutralizing capacity of an antacid tablet

Hydrochloric acid neutralizing capacity of commercial antacid tablets are summerized in

Table 1 and graphically depicted in Figure 2.

2. FTIR study of Pantafol-DSR

Interpretation of FTIR Spectra of Pantafol-DSR can be done as follows:

Spectral Compound Class

Pattern and intensity of Bond causing
region wave
Band Absorption
number cm-1
3950.22 Broad and low intensity - -
3934.78 Broad and low intensity - -
3901.99 Broad and low intensity - -

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3859.56 Broad and low intensity - -

3741.90 Broad and low intensity - -
3670.54 Sharp and moderate intensity O-H stretching Alcohol
3554.81 Broad and strong intensity O-H stretching Alcohol
3387.00 Broad and strong intensity O-H stretching Alcohol
3352.28 Broad and strong intensity O-H stretching Alcohol
3176.76 Broad and strong intensity O-H stretching Carboxylic acid
3022.45 Broad and strong intensity O-H stretching Carboxylic acid
2937.59 Broad and strong intensity O-H stretching Carboxylic acid
2825.72 Broad and strong intensity O-H stretching Carboxylic acid
2777.50 Broad and strong intensity O-H stretching Carboxylic acid
2634.76 Broad and strong intensity O-H stretching Carboxylic acid
2360.87 Sharp and moderate intensity - -
2266.36 Broad and low intensity - -
2135.20 Broad and low intensity C≡C stretching Alkyne
2071.55 Broad and low intensity - -
2038.76 Broad and low intensity - -
1915.31 Broad and low intensity C-H bending Aromatic compound
1865.17 Broad and low intensity C-H bending Aromatic compound
1826.59 Broad and low intensity C-H bending Aromatic compound

C=O stretching Conjugated acid

1701.22 Broad and strong intensity
C=O stretching Conjugated aldehyde
1622.13 Broad and low intensity - -
1591.27 Broad and low intensity - -
1485.19 Broad and moderate intensity - -
1456.26 Broad and moderate intensity - -
1381.03 Broad and low intensity - -
1309.67 Broad and low intensity - -
1273.02 Broad and low intensity - -

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1244.09 Broad and low intensity - -

1155.36 Broad and low intensity - -
1068.56 Broad and low intensity - -
995.27 Broad and low intensity - -
927.76 Broad and low intensity - -
862.18 Broad and low intensity - -
835.18 Broad and low intensity - -
758.02 Broad and low intensity - -
700.16 Broad and low intensity - -
657.73 Broad and low intensity - -
609.51 Broad and low intensity - -
547.78 Broad and low intensity - -

CONCLUSION

1. Determination of Hydrochloric acid neutralizing capacity of an antacid tablet

The higher the amount of Hydrocloric acid neutralized by Antacids, the better is the acid
neutralizing capacity. The result shows that the tablet Rantac 150 has highest acid neutralizing
capacity and PAN_L lowest acid neutralizing capacity value. Rantac 150 >Rabifeel – LS >
Omee-D > Pantafol-DSR > Rabesec-20 > NEXPRO-40 > Athzol-DSR > ACILOC 150 >
Gelusil > PAN-L. The back titration method used in this study is simple, inexpensive and can
be used in routine monitoring of the quality of Antacid tablets.

2. FTIR study of Pantafol-DSR

Interpretation of FTIR Spectra of Pantafol-DSR shows Presence of various functional groups
such as C-H bending - Aromatic compound; O-H stretching – Alcohol, Carboxylic acid, C=O
stretching - Conjugated acid, Conjugated aldehyde, C≡C stretching – Alkyne.

REFERENCES

1. ^ Jump up to:a b Internal Clinical Guidelines Team. (UK) (2014). Dyspepsia and Gastro-
Oesophageal Reflux Disease: Investigation and Management of Dyspepsia, Symptoms
Suggestive of Gastro-Oesophageal Reflux Disease, or Both. National Institute for Health
and Care Excellence: Clinical Guidelines. London: National Institute for Health and Care
Excellence (UK). PMID 25340236.
2. ^ Jump up to:a b Salisbury, Blake H.; Terrell, Jamie M. (2020), "Antacids", StatPearls,
Treasure Island (FL): StatPearls Publishing, PMID 30252305, retrieved 24
November 2020

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3. ^ "Aluminum hydroxide and magnesium carbonate Uses, Side Effects &

Warnings". Drugs.com. Retrieved 24 November 2020.
4. ^ Jump up to:a b c U.S. Department of Health & Human Services. Agency for Healthcare
Research and Quality 23 September 2011 Consumer Summary – Treatment Options for
GERD or Acid Reflux Disease: A Review of the Research for Adults Archived 11 October
2014 at the Wayback Machine
5. ^ Jump up to:a b Salisbury, Blake H.; Terrell, Jamie M. (2020), "Antacids", StatPearls,
Treasure Island (FL): StatPearls Publishing, PMID 30252305, retrieved 23
December 2020
6. ^ Graham, D. Y. (1982). "Pancreatic enzyme replacement: the effect of antacids or
cimetidine". Digestive Diseases and Sciences. 27 (6): 485–
490. doi:10.1007/BF01296725. ISSN 0163-2116. PMID 6282548. S2CID 10640940.
7. ^ Practice Guidelines for Preoperative Fasting and the Use of Pharmacologic Agents to
Reduce the Risk of Pulmonary Aspiration: Application to Healthy Patients Undergoing
Elective Procedures: An Updated Report by the American Society of Anesthesiologists
Task Force on Preoperative Fasting and the Use of Pharmacologic Agents to Reduce the
Risk of Pulmonary Aspiration. Anesthesiology. 2017 March; 126(3).
8. ^ George, J; Majeed, W; Mackenzie, IS; Macdonald, TM; Wei, L (26 November
2013). "Association between cardiovascular events and sodium-containing effervescent,
dispersible, and soluble drugs: nested case-control study". BMJ (Clinical Research
Ed.). 347: f6954. doi:10.1136/bmj.f6954. PMC 3898660. PMID 24284017.
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National Library of Medicine. Page last updated: 7 November 2014 Medline Plus: Taking
Antacids
10. ^ Texter, E. C. (1989). "A critical look at the clinical use of antacids in acid-peptic disease
and gastric acid rebound". The American Journal of Gastroenterology. 84 (2): 97–
108. ISSN 0002-9270. PMID 2644821.
11. ^ Hade, J. E.; Spiro, H. (1992). "Calcium and acid rebound: a reappraisal". Journal of
Clinical Gastroenterology. 15 (1): 37–44. doi:10.1097/00004836-199207000-
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12. ^ Bardhan, Karna Dev; Strugala, Vicki; Dettmar, Peter W. (2012). "Reflux Revisited:
Advancing the Role of Pepsin". International Journal of Otolaryngology. 2012:
646901. doi:10.1155/2012/646901. ISSN 1687-9201. PMC 3216344. PMID 22242022.
13. ^ Jump up to:a b c Ogawa, Ryuichi; Echizen, Hirotoshi (2011). "Clinically Significant Drug
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(2020). "A Systematic Review of Gastric Acid-Reducing Agent-Mediated Drug–Drug
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447–462. doi:10.1007/s40262-019-00844-3. ISSN 0312-
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17. ^ "Maalox Antacid Oral: Uses, Side Effects, Interactions, Pictures, Warnings &
Dosing". WebMD. Retrieved 24 June 2022.

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18. ^ Dubogrey, Ilya (2013). "Putting the Fizz into Formulation". European Pharmaceutical
Contractor. No. Autumn.
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21. ^ "Alka Seltzer Directions of use, Sodium & Aspirin content - Alka Seltzer relief from
Headaches, Migraine & Upset stomach". alkaseltzer.ie. Archived from the original on 29
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22. ^ Blair, G. T.; DeFraties, J. J. (2000). "Hydroxy Dicarboxylic Acids". Kirk-Othmer
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0471238966.
23. ^ Graham, David Y.; Smith, J. Lacey (1 March 1986). "Aspirin and the Stomach". Annals
of Internal Medicine. 104 (3): 390–398. doi:10.7326/0003-4819-104-3-390. ISSN 0003-
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24. Evaluation of Neutralizing Capacity of Different Commercial Brands of Antacid Tablets,
Abdu, K. and Abbagana, M., Chem Search Journal 6(2): 32 – 34, December, 2015
25. Evaluation of the Acid Neutralizing Capacity of Some Commercially Available Brands of
Antacid Tablets in Nigeria, Alalor CA, Avbunudiogba JA , Builders FP, Okpara LO, East
African Scholars Journal of Medical Sciences, Volume-2, Issue-1, January-2019, pp 12-16
26. Evaluation of acid neutralizing and buffering capacities of selected antacids in Ghana, Isaac
Ayensu, Samuel Oppong Bekoe, Joseph Kwasi Adu, Abena Amponsaa Brobbey, Scientific
African Volume 8, July 2020, e00347
27. Evaluation of the Effectiveness of Acid-Neutralizing Property of Traditional Antacids
commonly used in India, Divya J.O and Faseela Mohammed Rasheed, Journal of Scientific
Research, Volume 65, Issue 4, 2021
28. Evaluation of the acid-neutralizing capacity and other properties of antacids marketed in
Morocco, Mohamed Yafout, Hicham Elhorr, Ibrahim Sbai El Otmani, Youssef Khayati,
Medicine Pharmacy Reports, 2022 Jan; 95(1): 80–87.

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62

Investigation of Physico-chemical parameters of groundwater from

Barshitakli tehsil region, Akola district, Maharashtra, India

Dr. S. S Deshmukh1, Dr. Tejas R. Patil2

1,2
Ghulam Nabi Azad Arts, Commerce and Science College, Barshitakli, Dist. Akola.
Sant Gadge Baba Amravati University, Amravati.
[email protected]
ABSTRACT
This investigation determines the ground water quality of regions in Barshitakli tehsil,
Akola district in Maharashtra. Samples collected from various bore wells that were used for
drinking purposes. The physico-chemical parameters such as Electrical conductivity, pH, Total
hardness, Mg hardness, Ca hardness, Mg++ ions, Ca++ ions and Chloride ion were investigated
to determine the present condition of the groundwater quality during the periods from
December 2020 to December 2021 (consecutive post monsoon and pre monsoon). Overall
ground water quality of Barshitakli tehsil region is unsafe for human being. The groundwater
samples of bore well from Alanda and Januna village has the Turbidity beyond prescribed
limits of drinking water (IS:10500). The post monsoon sample shows increase in the values of
physio-chemical parameters of these two villages were due to higher erosion of soil. The
advanced farming and excessive use of chemical fertilizer and pesticides might has resulted in
the contamination of ground water.
Keywords: Physico-chemical, groundwater, Electrical conductivity, Turbidity, contamination.
INTRODUCTION
Potable water is also called as life and it has less abundance on the earth. Almost all
life processes on earth require water. The 71% of area on the earth’s crust is occupied by water
but earth’s 97% of water is present in the oceans which is too salty for drinking, growing crops
and for industrial purposes. The water to be describe as potable, the certain physical, chemical
and microbiological standards should be designed so as to ensure the water is potable and safe
for drinking (Tebutt, 1993 and Oladipo and Adeboye, 2015). Most of the industries discharge
their effluent without proper treatment into nearby open pits or pass them through unlined
channels, resulting in the contamination of ground water (Rao and Mamatha, 2004, Mahanta
et. al., 2015, Pande et. al., 2010). Potable water should be free from disease causing chemical
substances and micro-organisms to health (Ihekoronye and Ngoddy, 1985). The majority of
fresh water is existing in the ground in the form of soil moisture and beneath the ground in
aquifers.
The aim of present investigation is to study the groundwater samples collected from
bore well so as to understand the physico-chemical parameters of water in the Barshitakli tehsil
region, Akola district, Maharashtra.
STUDY AREA

Barshitakli is a taluka located in Akola district of Maharashtra. It is one of 7 Talukas of

Akola district. There are 159 villages in Barshitakli Taluka. The total area of Barshitakli is
812.97 sq.km with population density of 184 per sq.km.

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MATERIAL AND METHODS

The water samples from Barshitakli tehsil were collected from bore wells of ten
different villages. Water sample were collected in poly bottle during each consecutive month.
Physico- chemical parameters like water temperature, pH and transparency were recorded
immediately at the time of sample collection by using thermometer, portable digital pH meter
and transparency was measured with the help of Secchi disc respectively. The samples were
then transferred to laboratory for further analysis. The Parameters Such as TDS, Hardness and
chlorides were estimated in the Laboratory by using standard Methods as prescribed by APHA.

SAMPLE COLLECTION

Ground water samples were collected from ten (10) bore wells at various locations
within study area during pre and post monsoon season. Details of sampling locations are
along with their co-ordinates illustrated in Table1.

PHYSICO-CHEMICAL ANALYSIS OF GROUND WATER

The collected samples were analyzed for different physico-chemical parameters such
as pH, Electrical conductivity, Turbidity, TDS, Total hardness, Ca hardness, Mg hardness,
Ca ion, Mg ion, Chloride, and Temperature as per the standard methods (APHA, 1998) and
the results were compared with the Indian Standards (IS: 10500) for potable water.

RESULTS AND DISCUSSION

The water quality analysis of different ground water samples has been carried out for
pH, Electrical conductivity, TDS, Total hardness, Ca hardness, Mg hardness, Ca ion, Mg ion,
Chloride, and Temperature. The status of water quality of these ground water sources is given
in table 2.

Table 1: Sampling location and their coordinates

Sampling Location Area Location Co-ordinates
Code
GW1 Alanda 20.712456, 77.333320
GW2 Barshitakli 20.685510, 77.284157
GW3 Bhendi Mahal 20.666653, 77.268490
GW4 Chincholi Rudrayani 20.832091, 77.544832
GW5 Dagadparwa 20.730045, 77.404649
GW6 Dhaba 20.661896, 77.355489
GW7 Donad BK. 20.654725, 77.302891
GW8 Januna 20.694452, 77.254599
GW9 Kanheri 20.709671, 77.234655
GW10 Mahan 20.700926, 77.321669

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Table 2: Showing status of water quality of these ground water sources

Total Chloride Temp
Turbidity Conductance TDS Ca Mg
Area pH Hardness oC
(cm) (μmhos/cm) (mg/l) I Ion (mg/l)
code (mg/l)
on (mg/l)
(mg/l)
Pr Po Pr Po Pr Po Pr Po Pr Po Pr Po Pr Po Pr Po Pr Po
GW1 7.3 8.0 11.2 16.5 120 180 170 210 468 450 80 180 72 54 120 140 26 22
GW2 6.7 7.2 5.6 6.5 170 210 180 215 450 509 72 250 74 62 160 164 27 24
GW3 6.9 7.1 3.2 5.2 160 230 210 284 480 450 78 260 68 81 142 155 24 27
GW4 7.3 6.8 4.1 5.6 133 270 210 246 502 457 97 240 87 46 155 190 25 21
GW5 7.2 7.2 2.8 4.3 125 166 240 246 350 605 68 210 72 72 86 120 28 23
GW6 6.3 7.4 3.5 7.1 125 188 250 290 498 544 65 150 64 64 182 20 29 24
GW7 7.2 6.9 3.5 4.7 142 200 270 285 609 807 54 168 68 52 163 190 25 24
GW8 7.1 7.8 8.2 13.2 126 198 286 290 568 402 80 125 64 45 54 144 24 26
GW9 7.5 7.6 4.9 6.1 146 210 244 302 390 512 61 230 81 65 144 203 26 24
GW10 7.4 7.6 7.9 9.5 180 234 246 325 480 562 60 200 54 45 200 256 24 22
Mean 7.225 6.68 175.65 269.3 267.05 136.4 64.5 149.4 23.5
IS: 500- 250-
6.5- 8.5 5.0-10* ---------- 300-600 75-200* 30-100* --
10500 2000* 1000*
Pr- Pre-Monsoon, Po-Post Monsoon

Finding of the present investigation is as follows-

1. The pH value found permissible limit as per IS: 10500 of all samples.
2. Turbidity of samples from Alanda (GW1) and Januna (GW8) found 16.5 and 13.6
respectively which is the not in permissible limits as per IS: 10500.
3. Electrical conductivity varied between 120 to 180 μmhos/cm to 180 to 270 μmhos/cm
in pre and post monsoon season.
4. The same trend was observed in the case of total hardness of various ground water
sources. It varied from 170 to280 mg/l and 210 to 325 mg/l in pre and post monsoon
respectively.
5. TDS in all the samples were found to be within standard limits (IS:10500). In few
samples the ions of calcium have crossed the standard limit (IS: 10500) during post
monsoon season.
6. Chloride content of the ground water samples of hand pump were found to be within
limit during in the range pre and post monsoon season respectively.
7. Temperature of the various ground water samples were noted to be from 24-29 oC and
21-26 oC during pre and post monsoon season respectively.

CONCLUSION

Overall ground water quality of Barshitakli tehsil region is unsafe for human being. The
groundwater samples of bore well from Alanda and Januna village has the Turbidity beyond
prescribed limits of drinking water (IS:10500). The post monsoon sample shows increase in
the values of physio-chemical parameters of these two villages were due to higher erosion of
soil. The advanced farming and excessive use of chemical fertilizer and pesticides might has
resulted in the contamination of ground water.
Conflicts of interest: The authors stated that no conflicts of interest.

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REFERENCES

Ihekoronye, A. I., & Ngoddy, P. O. (1985). Integrated food science and technology for the
tropics. Macmillan.
Mahanta, C., Enmark, G., Nordborg, D., Sracek, O., Nath, B., Nickson, R. T., ... &
Bhattacharya, P. (2015). Hydrogeochemical controls on mobilization of arsenic in
groundwater of a part of Brahmaputra river floodplain, India. Journal of Hydrology: Regional
Studies, 4, 154-171.

Oladipo, I. C., & Adeboye, O. A. (2015). Physico-chemical and bacteriological analysis of

well water used for drinking and domestic purposes in Ogbomosho, Nigeria. International
Journal of Current Microbiology and Applied Sciences, 4(9), 136-145.

Pandey, V. P., Chapagain, S. K., & Kazama, F. (2010). Evaluation of groundwater

environment of Kathmandu Valley. Environmental Earth Sciences, 60, 1329-1342

Rao, S. M., & Mamatha, P. (2004). Water quality in sustainable water management. Current
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Tebutt, T. H. Y. (1983). Principles of quality control. Pergalmon, England, 235.

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63

Identification of confluence of multiple scaffolds required for Aurora

Kinase A inhibition

Vijay H. Masand1, S.D. Thakur2, M.M. Rathore1

1
Vidya Bharati Mahavidyalaya, Amravati, Maharashtra, India- 444 602
2
RDIK and NKD College, Badnera-Amravati, Maharashtra, India.

Abstract:
A vital signaling kinase with a significant function in cell division is Aurora Kinase A (AKA).
Therefore, inhibiting AKA is a desirable strategy to reduce cancer. Herein, we report a QSAR
(Quantitative Structure-Activity Relationships) analysis based on a set of 892 structurally
different AKA inhibitors to explore the features governing AKA inhibition. A QSAR model
with high statistical performance was created as per OECD (Organisation for Economic Co-
operation and Development) guidelines (R2tr = 0.794, Q2LOO = 0.787, R2ex = 0.778, CCCex
= 0.880). Predictive QSAR (external predictive ability) and Mechanistic QSAR (mechanistic
interpretations) are well-balanced in the recently developed QSAR model, which is based on a
set of eight variables. The present QSAR analysis is successful in identifying not only obscure
and obvious structural features, but also the reported and unpublished pharmacophoric
characteristics. The present research suggests that non-ring nitrogen atoms in combination with
planer nitrogen and non-ring oxygen must exist at a precise distance from one another.
Furthermore, the limit for simultaneous existence of sp2 oxygen with sp3 carbon and ring
oxygen is a new feature identified in the present work.

Introduction:
The mitotic machinery, also known as cell division mechanism, is profoundly regulated[1].
Any abnormality or defect in mitosis produces nondiploid DNA, which eventually leads to
cancer. The development of cancer chemotherapy drugs that specifically target centrosome
maturation and separation, mitotic spindle assembly, chromosomal segregation, and
cytokinesis has therefore attracted the attention of researchers. These processes involve the
crucial role of several signaling kinases, such as Aurora[2], Plk (Polo-like kinase), and Cdk
(cyclin-dependent kinase), to name a few. The Aurora kinase family of serine/threonine kinases
is essential for the reliable accomplishment of mitosis[3]. Therefore, these kinases have drawn
a lot of attention since their discovery in 1995 and the early identification of their expression
in human cancer tissue in 1998. The present involves QSAR analysis to identify important
structural features.
Methodology:
The structures were downloaded from ChEMBL (https://www.ebi.ac.uk/chembl/) for further
analysis. The next step involves conversion of SMILES notation to 3D-optimized structure
using MMFF94 (Cut-off: 0.01). Then, the molecular descriptors were calculated using
PyDescriptor[4] and PaDEL[5]. Before actual feature selection, descriptor pruning was
accomplished. After getting the reduced pool of descriptor GA-MLR was used for model
building. After that, the model was validated using different parameters.

Results and Discussion:

The eight parametric model along with its statistical validation is as following:

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Model-A:
Estimate Std. Error t value Pr(>|t|) Conf.Int.(+/-95%)
(Intercept) 7.521 0.0314 239.8884 0 0.0616
fringOH6B 0.334 0.0204 16.3689 0 0.0401
fnotringNplaN3B 1.1305 0.0453 24.9618 0 0.0889
fsp2Osp3C4B 0.4022 0.0338 11.8862 0 0.0664
fnotringOnotringN5B 0.4113 0.0546 7.527 0 0.1073
r_ringO_sp2O -0.9131 0.0796 -11.4699 0 0.1563
r_sp3C_sp2O -0.1813 0.0089 -20.431 0 0.0174
KRFPC992 -0.7937 0.0361 -21.9712 0 0.0709
KRFPC3612 -1.7261 0.0679 -25.4162 0 0.1333

Statistical parameters associated with Model-A:

S.N. Parameter Value S.N. Parameter Value

1. R2_tr 0.7941 20. RMSE_ex 0.5129
2. Adj-R2 0.7918 21. PRESS_ex 47.0818
3. F(8-704) 339.4136 22. Q2F1 0.7674
4. RSS_tr 154.2898 23. Q2F2 0.7661
5. MSE_tr 0.2164 24. Q2F3 0.7497
6. RMSE_tr 0.4652 25. MAE_ex 0.4328
7. MAE_tr 0.3911 26. K 1.0069
8. s 0.4681 27. K_prime 0.9881
9. AIC 952.0537 28. R2ext 0.7782
10. BIC 997.7485 29. CCC_ex 0.8802
11. CCC_tr 0.8852 30. r2m_ExPy 0.6736
12. Q2_cv 0.7872 31. r2m_EyPx 0.701
13. RMSE_cv 0.473 32. R2o 0.7683
14. MSE_cv 0.2237 33. R2o_dash 0.7601
15. PRESS_cv 159.498 34. Clos_dash 0.0232
16. MAE_cv 0.3969 35. Clos 0.0126
17. R2_Yscr 0.011 36. r2m_avg 0.6873
18. MSE_ex 0.263 37. r2m_delta 0.0273
19. Training Set Compounds 713 38. Prediction Set Compounds 179

The details of descriptors are as following:

Descriptor Details Software used

fringOH6B Frequency of occurrence of hydrogen exactly at PyDescriptor
six bonds from ring oxygen
fnotringNplaN3B Frequency of occurrence of planer nitrogen PyDescriptor
exactly at three bonds from non-ring nitrogen
fsp2Osp3C4B Frequency of occurrence of sp3-hybridized PyDescriptor
carbon exactly at four bonds from sp2-
hybridized oxygen

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fnotringOnotringN5B Frequency of occurrence of non-ring nitrogen PyDescriptor

exactly at five bonds from non-ring oxygen
r_ringO_sp2O Ratio of ring oxygen to sp2-hybridized oxygen PyDescriptor
r_sp3C_sp2O Ratio of sp3-carbon to sp2-hybridized oxygen PyDescriptor
KRFPC992 [!#1][NH]c1[cH][cH][cH]c([!#1])[cH]1 PaDEL
Meta-substituted aniline ring
KRFPC3612 Cc1ccn[nH]1 PaDEL
3-substituted pyrazole moiety

Model-A contains eight molecular descriptors covering broad chemical space. Four of the eight
descriptors, namely fringOH6B, fnotringNplaN3B, fsp2Osp3C4B, and fnotringOnotringN5B,
have positive coefficients in model-A, suggesting that augmenting their value could result in a
better inhibition of AKA. The opposite is true for the four remaining descriptors,
r_ringO_sp2O, r_sp3C_sp2O, KRFPC992, and KRFPC3612. It is to be noted that every
molecular descriptor is a mathematical (or numeric) representation of one or more
pharmacophoric features that control the biological profile. Further, the final biological activity
(say IC50, Ki, etc.) of a molecule varies with multiple pharmacophore features and not by a
single feature. In other words, the biological activity is the result of a confluence of various
structural characteristics and some unidentified elements. Some characteristics are in charge of
enhancing the desired pharmacological action, while others are responsible for reversing it. It
is thought that the biological activity is determined concurrently by two or more
pharmacophoric groups (Pharmacophore synergism).
Conclusion:
The present research suggests that non-ring nitrogen atoms in combination with planer nitrogen
and non-ring oxygen must exist at a precise distance from one another. Furthermore, the limit
for simultaneous existence of sp2 oxygen with sp3 carbon and ring oxygen is a new feature
identified in the present work. In conclusion, the AKA inhibitory activity is associated with a
group of structural features.
References:
1. Du R, Huang C, Liu K, et al. Targeting AURKA in Cancer: molecular mechanisms and
opportunities for Cancer therapy. Molecular Cancer. 2021;20(1). doi: 10.1186/s12943-020-
01305-3.
2. Bavetsias V, Linardopoulos S. Aurora Kinase Inhibitors: Current Status and Outlook. Frontiers
in Oncology. 2015;5. doi: 10.3389/fonc.2015.00278.
3. Borisa AC, Bhatt HG. A comprehensive review on Aurora kinase: Small molecule inhibitors
and clinical trial studies. European Journal of Medicinal Chemistry. 2017;140:1-19. doi:
10.1016/j.ejmech.2017.08.045.
4. Masand VH, Rastija V. PyDescriptor : A new PyMOL plugin for calculating thousands of easily
understandable molecular descriptors. Chemometrics and Intelligent Laboratory Systems.
2017;169:12-18. doi: 10.1016/j.chemolab.2017.08.003.
5. Yap CW. PaDEL‐descriptor: An open source software to calculate molecular descriptors and
fingerprints. Journal of Computational Chemistry. 2010;32(7):1466-1474. doi:
10.1002/jcc.21707.

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64

Synthesis, Characterization and Biological Evaluation of 4-(4-Bromo-1-

Hydroxy Naphthalen-2-Yl)-6-(4-Hydroxy Phenyl)-5,6-Dihydropyrimidine-
2(1h)-One

Dr. V. M. Sherekar*1, Mr. Nilesh S. Padole2

Assistant Professor, Department of Chemistry, Vinayak Vindhyan Mahavidyalaya Nandgaon (Kh), Dist.
Amravati (M.S), India.
Head and Assistant Professor, Department of Chemistry, Vinayak Vidnyan Mahavidyalaya, Nandgaon
Khandeshwar, Dist. Amravati (M.S.), India.

ABSTRACT: -
1-(4-Bromo-1-hydroxynaphthalen-2-yl)-ethan-1-one was prepared by refluxing 4-
bromonaphthalen-1-ol with glacial acetic acid in presence of fused ZnCl2. By condensing 1-(4-bromo-
1-hydroxynaphthalen-2-yl)-ethan-1-ones with 4-Hydroxy benzaldehyde, to prepared by 1-(4- bromo -
1- hydroxynaphthalen-2-yl)-3-(4-Hydroxy phenyl)-prop-2-en-1-one were synthesized.1-(4- bromo -1-
hydroxynaphthalen-2-yl)-3-(4-Hydroxy phenyl)-prop-2-en-1-one, urea and concentrated HCl in DMF
were added and refluxed. Cool and pour in crushed ice. Treat it with cold NH4OH solution to obtain
titled compounds. The compounds thus synthesized have been characterized by physical and spectral
data. All of these titled synthesized compounds have been screened for antimicrobial study and are
found to possess excellent antimicrobial activities.
KEYWORDS: - antimicrobial activities, cold NH4OH solution, concentrated HCl in DMF.
INTRODUCTION: -
Dihydropyrimidin-2(1H)-one are classified as hetero-cyclic compound and
containing pyrimidine ring which is containing two nitrogen atoms in the six-member ring.
Dihydropyrimidine are the most important heterocyclic ring system which play an important role in the
synthesis of DNA and RNA. They widely using multi-component reaction like Biginelli reaction. In
the field of heterocyclic chemistry dihydropyrimidine-2(1H)-one was synthesized through the one -pot
condensation of an aromatic aldehyde and urea in the presence of the basic [1-3]. Dihydropyrimidines
are one of the important heterocyclic compounds, which are of interest due to its efficiency towards
various pharmacological uses [4]. The synthesis of the dihydropyridine and their derivatives increasing
tremendously significant because they generally show diverse medicinal properties [5]. Newly
researcher goal dihydropyrimidine derivatives modulated heat shock responses and have
neuroprotective responses such like that optimized for their ability to modulate cellular stress responses
based on favorable toxicological data [6]. Many aryls substituted dihydropyrimidine-2-one are found
to exhibited biological activities [7]. Many reports exploring in Vivo and in Vitro dihydropymidine-2-
one derivatives show variety of pharmacological activities such as active and safe tumor anti-initiating
and multi-potent blocking agent [8], anxiolytic [9], antihypertensive agents [10], anticonvulsant [11],
anticancer [12], analgesic activities [13], anti-bacterial [14], channel blockers [15], anti-HIV [16].
Their efforts are quite significant in literature hence considering the scope of
dihydropyrimidine derivatives we have synthesized novel4-(4-bromo-1-hydroxynaphthalen-2-yl)-6-(4-
hydroxy phenyl)-5,6-dihydropyrimidine-2(1h)-one from 4- bromonaphthalen-1-ol and studied for their
biological activities.
MATERIALS AND METHOD: -
Synthesis of 1-(4-Bromo-1-hydroxynaphthalen-2-yl)-ethan-1-one.

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1-(4-Bromo-1-hydroxynaphthalen-2-yl) ethan-1-one was prepared by modified Nenchis method in

which 4-bromo- naphthalen-1-ol was refluxed with glacial acidic acid in presence of fused ZnCl2.
Synthesis of 1-(4-Bromo-1-hydroxynaphthalen-2-yl)-3-(4-Hydroxy phenyl)-prop-2-en-1-one.
1-(4-Bromo-1-hydroxynaphthalen-2-yl)-3-(4-Hydroxy phenyl)-prop-2-en-1-one were synthesized
from 1-(4-Bromo-1-hydroxynaphthalen-2-yl) ethan-1-one by condensing it with 4-Hydroxy
benzaldehyde were added in ethanol solvent and KOH mixture.
Synthesis of 4-(4-Bromo-1-hydroxy naphthalen-2-yl)-6-(4-Hydroxy phenyl)-5,6-
dihydropyrimidine-2(1H)-one.
4-(4-Bromo-1-hydroxy naphthalen-2-yl)-6-(4-methoxy phenyl)-5,6-dihydropyrimidine-
2(1H)-one were prepared from 1-(4-Bromo-1-hydroxynaphthalen-2-yl)-3-(4-Hydroxy
phenyl)-prop-2-en-1-one was reflux with urea and concentrated HCl in DMF. It was
then treated with cold NH4OH.

In present work the compounds under investigation are: -

Compound 1: 4-(4-Bromo-1-hydroxy naphthalen-2-yl)-6-(4-hydroxy phenyl)-5,6-dihydropyrimidine-
2(1H)-one.
Compound 2: 4-(4-Bromo-1-hydroxy naphthalen-2-yl)-6-(3, 4-hydroxy phenyl)-5,6-dihydropyrimidine-
2(1H)-one.
Compound 3: 4-(4-Bromo-1-hydroxy naphthalen-2-yl)-6-(3-Hydroxy phenyl)-5,6-dihydropyrimidine-
2(1H)-one.
Compound 4: 4-(4-Bromo-1-hydroxy naphthalen-2-yl)-6-(4-Hydroxy phenyl)-5,6-dihydropyrimidine-
2(1H)-one.
SCHEME: -

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Sr. Compound R1 R2 Molecular Melting % % Nitrogen R.F

no formula Point Yield Value
no 0
C
Found Calculated

1 1 - -H C17H17N2O2Br 2590C 45% 6.64 6.61 0.55

OCH3

2 2 - OCH3 C17H19N2O4Br 2250C 48% 6.21 6.74 0.59

OCH3
3 3 -H -OH C17H15N2OBr 2280C 45% 6.89 6.84 0.55

4 4 -OH -H C17H15N2O2Br 2690C 51% 5.84 5.85 0.58

Table 1. PHYSICAL DATA OF SYNTHESIZED COMPOUNDS

SPECTRAL ANALYSIS: -
IR(max) (cm-1): 1624 (C=O, str), 3346 (NH, str), 1568 (C=N), 1171 (C-O-C),758(monosubstituted
Benzene
NMR ( ppm): 1.3-1.8 (m, 2H, -CH2 of pyrimidine), 10.30 (s, 1H, -OH),3.61 (s, 3H, -OH),2.51 (s, 3H,
CH3,)
ANTIMICROBIAL STUDIES: -
All above synthesized 4-(4-Bromo-1-hydroxy naphthalen-2-yl)-6-(4-hydroxy phenyl)-5,6-
dihydropyrimidine-2(1H)-one have been studied for their antimicrobial activity against Escherichia coli,
Proteus mirabilis, Staphylococcus aureus, Pseudomonas aeruginosa. The culture of each species was
incubated at 370C and the zone of inhibition was measured after 24 hr. Results are tabulated in Table.
Most of these compounds were found active

Sr. Compound Antimicrobial Activity

no Number
E-coli Proteus mirabilis Staphylococcus Pseudomonas
aureus aeruginosa

1 1 17 18 15 10

2 2 18 10 16 14

3 3 16 13 14 15

4 4 15 14 12 13

Strongly active, range 15-19 Weakly active, range 7-10 mm, moderately active, range 11-14mm,
Inactive, -

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CONCLUSION: -
Thus, from above results it was observed that these heterocyclic compounds were found effective
against Escherichia coli, Proteus mirabilis, Staphylococcus aureus, Pseudomonas aeruginosa. So those
compounds can be easily be used for the treatment of diseases caused by test pathogens, only when they
do not have toxic and other side effects.

REFERENCE: -

1. MF Custodio; CC Allane et al. Z. Kristalloger, 2019, 234 (10), 657-669.

2. P Jain; A Patil.World J. Pharm. Res.2018, 7(11), 410-427.
3. V Garg: D Jindal; R Singh. TJPLS Journal, 2020, 7, 6, 8-12.
4. A Huseynzada et al; Royal Society of Chemistry, 2021, 11, 11, 6312-6329
5. A Malah; Z Mahmoud; H H Salem; A Abdou; M Soliman; R Hassan. Green Chemistry
Letters and Reviews, 2021,14, 2, 220-232.
6. K Agnes et al. J Alzheimers Dis, 2016, 53 (2), 557-571.
7. VM Sherekar; SE Bhandarkar. Am. J. PharmTech Res.2016, 6(5), 560-56.
8. VM Sherekar; SE Bhandarkar;World J. Pharm. Res.2016, 2(6), 275-279.
9. B Padmashali; BN Chidananda; B Govindappa; S Basavaraj. J. Appl. Pharm. Science.2019,
9(5), 133-140.
10. F Alam; R Amin. IJPSR, 2020, 32, 10, 46-55.
11. V A Adol et al. Material Science Research India, 2020, 17, 1, 13-26.
12. R Al-Saheb et al. Bioscience Reports, 2020, 40, 9, 1-13.
13. R Kaur; S Chaudhary; K Kumar; MK Gupta; RK Rawal. Eur J Med Chem.2017, 132,108-134.
14. AY Vargas; HA Rojas; GP Romanelli; JJ Martinez. Green Process Synth.2017,6(4), 377-384.
15. R M Keshk; B M Izzularab. Current Organic Synthesis, 2021, 18, 0, 1-10.
16. N Podilla; C Tirthankar. JAOPR, 2018, 6, 1, 11-15.

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65

Efficient Synthesis of Salicylaldehyde-Based Schiff Bases Using

Hydrothermal Sonication: A Green Approach

Vivek Ramkrushna Matea*, Datta Anandrao Patilb, Kalpana B. Gawandec,

Sanjay R. Thakarec, and Rajkumar Uddhavrao Pokalwalb
a
Department of Chemistry, Yashvantrao Chavan College, Mangrulpir, Washim, Maharashtra, India.
b
Department of Chemistry, Degloor College, Degloor, Nanded, Maharashtra, India.
c
Department of Chemistry, Institute of Science, Nagpur, Maharashtra, India.

This study presents a novel, efficient, and environmentally friendly procedure for the synthesis
of a series of salicylaldehyde-based Schiff bases utilizing hydrothermal sonication. The
procedure involves the condensation of salicylaldehyde with various aromatic amines in water
under hydrothermal sonication conditions. Through meticulous selection of the solvent and
reaction conditions, the final products are obtained in excellent yields in a single-step
procedure. Comparative experiments under thermal conditions yielded lower yields and
necessitated more laborious work-up processes. The hydrothermal sonication method
demonstrates several advantages, including reduced reaction time, increased conversion,
minimized waste production, and favorable yields. The synthesized compounds' structures
were confirmed through the analysis of Infrared (IR), Proton Nuclear Magnetic Resonance
(1HNMR and C13NMR), and Mass Spectra data. This research highlights the efficiency and
environmental benefits of the hydrothermal sonication method in synthesizing salicylaldehyde-
based Schiff bases.

Introduction
The increasing focus on the environmental impact of chemicals has prompted a drive towards
cleaner and more eco-friendly synthetic processes. Stringent regulations are geared towards
promoting sustainability by minimizing waste generation, eschewing the use of auxiliary
substances such as organic solvents and additional reagents, and reducing energy demands.
The utilization of water as the reaction medium is particularly advantageous in this context.
Firstly, water is cost-effective, nonflammable, non-toxic, and poses minimal safety concerns
[1, 3]. Secondly, its distinct physical and chemical properties often augment reactivity or
selectivity, which may not be attainable with organic solvents. Lastly, employing water
obviates the necessity to dry substrates or reagents prior to use, thereby diminishing or
eliminating the consumption of drying agents, energy, and time.

The synthesis of Schiff bases traditionally involves the condensation of a primary amine with
an aldehyde, often employing organic solvents such as methanol, tetrahydrofuran (THF), and
1,2-dichloroethane (DCE) [4]. Additionally, solvent-free methods utilizing hydrothermal
sonication have been reported for the synthesis of Schiff bases [5]. A comparative analysis of
these synthesis methods for simple Schiff bases revealed that hydrothermal sonication
irradiation represents the most straightforward approach [6].

The use of Schiff base compounds as ligands is widespread due to their facile formation and
diverse coordination chemistry, enabling their application as catalysts in various asymmetric
reactions [7, 8]. Notably, Schiff base complexes derived from salicylaldehyde, 2-aminol, and
3,4-thiadiazole have been evaluated for their antibacterial activity against a range of bacterial
strains, including Escherichia coli, Staphylococcus aureus, and Pseudomonas aeruginosa [9].

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It was observed that the antibacterial efficacy of these Schiff bases was enhanced through
chelation/complexation, demonstrating significant activity against the tested bacterial strains
and presenting new avenues in combating antibiotic-resistant strains. In light of these
discoveries, this manuscript presents the hydrothermal sonication-assisted synthesis of
salicylaldehyde-based Schiff bases in aqueous media.

The synthesis of Schiff bases has been extensively documented in the literature [10–13].
However, the reported methods often entail a laborious process utilizing methanol or ethanol
as solvents, leading to prolonged reaction times. Typically, the synthesis of Schiff bases
through condensation reactions necessitates stringent conditions, including the use of a Dean
Stark apparatus, catalysts, elevated temperatures, and extended reaction durations [14].
Notably, the significance of the fluorinated Schiff bases discussed in this manuscript is equally
noteworthy. This significance stems from the influence of fluorine substitution on inter- and
intramolecular forces, which affect ligand binding and introduce receptor subtype selectivity
in biological systems [15, 16]. The breadth and universality of this synthetic process are
demonstrated with respect to salicylaldehyde and various fluorinated amines.

In conclusion, we introduce a sustainable and effective approach for synthesizing Schiff bases
in aqueous media, as illustrated in Scheme 1. The uncomplicated workup, convenient
conditions, swift reaction kinetics, high yields, and reaction specificity collectively enhance
the appeal of the current methodology.

Experimental
The purity of the compound was assessed using silica-gel-coated aluminum plates (Merck).
Melting points were determined in open capillary tubes. Infrared (IR) spectra were recorded
with KBr on a Perkin Elmer Spectrum RX-1 FTIR spectrophotometer. Proton nuclear magnetic
resonance (1H-NMR) and carbon-13 nuclear magnetic resonance (13C-NMR) spectra were
acquired on a Jeol JNM-ECX400P instrument operating at 400 MHz. Hydrothermal
ultrasonication irradiations were conducted in a hydrothermal sonication synthesizer operating
at 37 KHz using Hielscher instrument. All chemicals utilized were of analytical grade.

Conventional Method
Schiff bases were synthesized through the condensation of salicylaldehyde (0.005 mol) with
various aromatic amines (a–f) (0.005 mol) in 10 mL of water. The reaction mixture was stirred
at ambient temperature, and the progress of the reaction was monitored using thin-layer
chromatography (TLC). Upon completion of the reaction, the product was isolated, filtered,
dried, and subsequently recrystallized using methanol.

Hydrothermal ultra sonication method

The hydrothermal ultrasonication synthesis method involves subjecting the reactants to high-
frequency ultrasound, generating intense sound waves that stimulate chemical reactions. In this
sonochemical approach, the reactants are placed in a sonication vessel, exposed to high-
frequency ultrasound, leading to localized heating, elevated pressures, and increased
concentrations of reactive species, facilitating chemical transformations and yielding the
desired Schiff base product.

A mixture comprising salicylaldehyde (0.005 mol) and substituted aromatic amines (2a–f)
(0.005 mol) in 10 mL of water was introduced into a hydrothermal sonication tube. The
contents underwent hydrothermal sonication irradiation at 37 KHz for a duration of 1 hour. The
progress of the reaction was monitored via thin-layer chromatography (TLC). Upon completion

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of the reaction, a solid product formed in the reaction mixture, which was subsequently filtered
and recrystallized using methanol. The recrystallization process yielded the title compounds in
the form of solid crystals.

Results and Discussion

A novel environmentally friendly method utilizing water as a solvent for the synthesis of Schiff
bases has been successfully developed, as depicted in the schematic representation in Scheme
1. The optimization of %yield was investigated at various temperatures, revealing an optimal
yield at 70 °C. Beyond this temperature, there was no significant change in yield, as
summarized in Table 1. The results indicate that at 30 °C, the yield is 5.5%, and with an increase
in temperature to 70 °C, the yield significantly improves to 53.5%. This tenfold increase in
activity may be attributed to enhanced solubility, increased agitation, and a reduction in
intermolecular distances. Consequently, the synthesis of other products (b–f) was carried out
under the same reaction conditions, maintaining a temperature of 70 °C. The data suggests that
the environmentally friendly method using water as a solvent, particularly at a temperature of
70 °C, significantly enhances the %yield for the synthesis of Schiff bases.

Table 1: Comparison of yield of Schiff base (a) at different temperatures under Hydrothermal
ultra sonication.

Compound Temperature (°C) %Yield

3a 30 5.5
3a 35 7.5
3a 40 9.5
3a 45 10.0
3a 50 13.6
3a 55 19.5
3a 60 35.4
3a 65 46.2
3a 70 53.5
3a 75 55.1
3a 80 55.5
Reaction Conditions :- Sonication 37 KHz, Time 1 hr, solvent water, and aldehyde : base
concentration 1:1 moles.
The impact of different compositions on %yield is detailed in Table 2. A variation in aldehyde
and base composition resulted in an increase in %yield from 45.5 to 54.5. This observed
increase is likely attributed to the stoichiometric composition of salicylaldehyde and
substituted aromatic amines. The optimal %yield of 53.5 was achieved for a composition ratio
of 1:1. Consequently, the same composition was adopted for the synthesis of other compounds
(b-f). It's clear that the composition ratio of salicylaldehyde and substituted aromatic amines
played a significant role in achieving the optimal %yield of 53.5. This information can be
valuable for further experiments or scale up applications.
Table 2: Effect of composition of aldehyde and base for the formation of Schiff base (a)

Composition %Yield
Aldehyde (mole) Base (mole)

1 0.75 45.5

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1 1.00 53.5
1 1.25 54.1
1 1.50 54.5

Reaction Conditions :- Temperature 70 °C, sonication 37 KHz, Time 1 hr, solvent water.
The comparative result analysis of the hydrothermal ultrasonication method is presented in
Table 3. The results obtained with this method were significantly higher, by almost 50%,
compared to the conventional hydrothermal method. This suggests that the hydrothermal
ultrasonication method, by enhancing the contact time of reactants, leads to a higher %yield.
Consequently, this highlights the advantageous nature of the hydrothermal ultrasonication
synthesis for the preparation of various Schiff bases. The data indicates that the hydrothermal
ultrasonication method is a more effective approach for the synthesis of Schiff bases, resulting
in a substantial increase in %yield compared to the conventional hydrothermal method. This
insight could be valuable for researchers and practitioners in the field of chemical synthesis.

Table 3: Comparative study of hydrothermal ultra sonication and conventional method.

Compound % Yield

Hydrothermal Conventional
ultra sonication
a 53.5 21.0
b 85.6 37.9
c 81.3 37.5
d 80.4 37.5
e 78.4 39.8
f 80.5 32.5

Reaction Conditions :- Temperature 70 °C, sonication 37 KHz, Time 1 hr, solvent water and
aldehyde : base concentration 1:1 moles.

Product Characterization Data

(a) (E)-2-(((2,4-dimethylphenyl)imino)methyl)phenol, colorless solid, M.P. 91- 93°C, IR

(KBr, cm−1): 2927.60, 2700, 1615.96, 1490.71, 1282.72, 1144.23, 1HNMR (400 MHz,
CDCl3) ( ppm): 8.87 (1CH, s), 7.66 (1H, d), 7.08 (2H, d), 70.2 (1H, s), 7.01 (1H, m),
2.34 (6H, m). 13C-NMR (CDCl3) ( ppm): 161.1, 160.0, 132.4, 132.1, 121.4, 120.5,
117.8, 148.1, 132.2, 128.7, 128.7, 127.3, 122.1, 21.6, 18.6, . M/S: 226.12 (M + 1).
(b) 2-[(4-Chloro-phenylimino)-methyl]-phenol [10]. Yellow solid, M.p. 95–98°C, IR
(KBr, cm−1): 2920.94, 1620.86, 1487.51, 1278.31, 1134.17, 1HNMR (400 MHz,
CDCl3):  ppm = 13.06 (1CH, s), 8.88 (1H, s), 7.67 (2H, d), 7.57 (1H, m), 7.49 (1H, t),
6.70 (3H, m). 13C-NMR (CDCl3) ( ppm): 159.21, 164.71, 163.28, 158.72, 138.09,
134.02, 133.01, 122.08, 120.72, 119.02, 117.41, 112.01, 105.05. M/S: 234.8 (M + 1).
(c) 2-[(4-Chloro-phenylimino)-methyl]-phenol [11]. Golden solid, M.p. 70–73°C, IR
(KBr, cm−1): 2950.53, 1617.09, 1474.16, 1272.73, 841.22, 1HNMR (400 MHz,
CDCl3): =  12.92 (1 H, s), 8.41 (1 H, s), 7.64 (2 H, d), 7.51 (2 H, d), 7.52 (2 H, dd),
7.34 (1 H, d), 7.02 (1 H, t). 13C-NMR (CDCl3) ( ppm): 163.07, 162.08, 148.01,
134.01, 133.32, 132.27, 132.18, 129.11, 123.03, 120.07, 119.01, 118.10. M/S: 232.84
(M+ 1).

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(d) 2-[(2,3,4,5-Tetrafluoro-phenylimino)-methyl]-phenol [12]. Greenish solid, M.p. 141–

143°C, IR (KBr, cm−1): 2931.04,1618.71, 1467.17, 1040.21, 775.99. 1HNMR (400
MHz, CDCl3): ppm = (1H, s), 8.41 (1H, s), 7.34 (2H, dd), 7.16 (1H, d), 6.91 (2H, m).
13C-NMR (CDCl3) (ppm): 164.19, 159.30, 139.25, 134.99, 132.55, 131.72, 129.95,
129.66, 117.83, 117.49, 116.58, 115.61, 101.07. M/S: 270.27 (M + 1).
(e) 2-[(4-Bromo-phenylimino)-methyl]-phenol [11, 12]. Greenish solid, M.p. 97–99°C, IR
(KBr, cm−1): 2911.15, 1616.74, 1480.31, 1271.05, 837.16, 763.41, 1HNMR (400 MHz,
CDCl3): ppm = (1H, s), 8.55 (1H, s), 7.51 (2H, d), 7.49 (2H, m), 7.11 (2H, d), 7.12
(1H, d), 6.97 (1H, t). 13C-NMR (CDCl3) (ppm): 161.99, 161.23, 147.67, 133.48,
132.44, 132.79, 122.19, 120.34, 119.41, 119.50, 117.62. M/S: 276.1 (M + 1).
(f) 2-[(4-Fluoro-phenylimino)-methyl]-phenol [13]. Yellow solid, M.p. 71–73°C, IR (KBr,
cm−1): 2927.88, 1614.71, 1494.00, 1260.05, 830.04, 1HNMR (400 MHz, CDCl3): 
ppm= 13.01 (1H, s), 8.58 (1H, s), 7.37 (2H, m), 7.26 (2H, m), 7.12 (2H, m), 7.03 (1H,
d), 6.94 (1H, t). 13C-NMR (CDCl3) ppm= 163.40, 161.08, 159.17, 142.01, 131.08,
128.45, 121.02.

Conclusions

In summary, the synthesis of Schiff bases through condensation reactions using both
conventional hydrothermal and hydrothermal ultrasonication methods was investigated, with
the latter proving to be the more efficient approach. The tenfold increase in activity observed
with an increase in temperature from 30 to 70 °C can be attributed to enhanced solubility,
increased agitation, and a reduction in intermolecular distances. The highest %yield, reaching
53.5%, was achieved with a composition of salicylaldehyde and the base at a temperature of
70 °C. The optimal composition of salicylaldehyde and substituted aromatic amines was found
to be 1:1 for maximum %yield. Subsequently, the synthesis of other products (b–f) was carried
out with a composition ratio of 1:1, maintaining a temperature of 70 °C. The data obtained
indicates that the hydrothermal ultrasonication method is a more effective approach for the
synthesis of Schiff bases, resulting in a substantial increase in %yield compared to the

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conventional hydrothermal method. This insight holds significance for researchers and
practitioners in the field of chemical synthesis.

References

1) S. Bhagat, N. Sharma, and T. Singh Chundawat, “Synthesis of Some Salicylaldehyde-Based Schiff

Bases in Aqueous Media” Journal of Chemistry, Vol. 2013, Article ID 909217, pp. 1-4. 2013.
2) C. J. Li, “Organic reactions in aqueous media with a focus on carbon-carbon bond formations: a
decade update,” Chemical Reviews, vol. 105, no. 8, pp. 3095–3165, 2005.
3) H. Yorimitsu, H. Shinokubo, and K. Oshima, “Synthetic radical reactions in aqueous media,”
Synlett, no. 5, pp. 674–686, 2002.
4) U. M. Lindstrom, “Stereoselective organic reactions in water,” Chemical Reviews, vol. 102, no. 8,
pp. 2751–2772, 2002.
5) A. F. Abdel-Magid, K. G. Carson, B. D. Harris, C. A. Maryanoff, and R. D. Shah, “Reductive
amination of aldehydes and ketones with sodium triacetoxyborohydride. Studies on direct and
indirect reductive amination procedures,” Journal of Organic Chemistry, vol. 61, no. 11, pp. 3849–
3862, 1996.
6) -base without solvent
under microwave irradiation,” Chinese Chemical Letters, vol. 13, no. 1, pp. 3–6, 2002.
7) Z. Yang and P. Sun, “Compare of three ways of synthesis of simple Schiff bas,” Molbank, vol.
2006, no. 6, Article ID M514, 2006.
8) S. C. Gagieva, T. A. Sukhova, D. �. Savinov et al., “New �uorinecontaining bissalicylidenimine-
titanium complexes for ole�n polymerization,” Journal of Applied Polymer Science, vol. 95, no. 5,
pp. 1040–1049, 2005.
9) A. Patti, S. Pedotti, F. P. Ballistreri, and G. T. Sfrazzetto, “Synthesis and characterization of some
chiral metal-salen complexes bearing a ferrocenophane substituent,” Molecules, vol. 14, no. 11, pp.
4312–4325, 2009.
10) Z. H. Chohan, M. F. Jaffery, and C. T. Supuran, “Antibacterial Co(II), Cu(II), Ni(II) and Zn(II)
complexes of thiadiazoles Schiff bases,” Metal-Based Drugs, vol. 8, no. 2, pp. 95–101, 2001.
11) D. P. Song, Y. G. Li, R. Lu, N. H. Hu, and Y. S. Li, “Synthesis and characterization of novel neutral
nickel complexes bearing fluorinated alicylaldiminato ligands and their catalytic behavior for
vinylic polymerization of norbornene,” Applied Organometallic Chemistry, vol. 22, no. 6, pp. 333–
340, 2008.
12) J. S. Bennett, K. L. Charles, M. R. Miner et al., “Ethyl lactate as a tunable solvent for the synthesis
of aryl aldimines,” Green Chemistry, vol. 11, no. 2, pp. 166–168, 2009.
13) W. Qingming, M. Zhu, R. Zhu et al., “Exploration of - aminophosphonate N-derivatives as novel,
potent and selective inhibitors of protein tyrosine phosphatases,” European Journal of Medicinal
Chemistry, vol. 49, pp. 354–364, 2012.
14) P. Kathirgamanathan, S. Surendrakumar, J. Antipan-Lara et al., “Novel lithium Schiff-base cluster
complexes as electron injectors synthesis, crystal structure, thin film characterisation and their
performance in OLEDs,” Journal of Materials Chemistry, vol. 22, no. 13, pp. 6104–6116, 2012.
15) S. Jie and S. Zhang, “2-arylimino-9-phenyl-1,10-phenanthrolinyl-iron, -cobalt and -nickel
complexes: synthesis, characterization and ethylene oligomerization behavior,” European Journal
of Inorganic Chemistry, vol. 2007, no. 35, pp. 5584–5598, 2007.
16) M. Rowley, D. J. Hallett, S. Goodacre et al., “3-(4-fluoropiperidin-3-yl)-2-phenylindoles as high
affinity, selective, and orally bioavailable h5-HT2a receptor antagonists,” Journal of Medicinal
Chemistry, vol. 44, no. 10, pp. 1603–1614, 2001.
17) S. Purser, R. M. Peter, S. Swallow et al., “Fluorine in medicinal chemistry,” Chemical Society
Reviews, vol. 37, no. 2, pp. 320–330, 2008.

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66

Viscometric Technique for Evaluation

of 1-Phenyl-3-[4-(2-Ethylimino-4-T-Butylimino-1,3,5-Dithiazino)Amino-
Phenyl]Prop-2-Ene-1-One in Ethanol-Water Mixture

S.S. Padhen1*, A.B. Wadekar2

1- Department of Chemistry, Rajarshee Shahu Science College Chandur Railway, Dist.-Amravati, 444 904.
Maharashtra, India.
2- Department of Chemistry, Shri. Dnyaneshwar Maskuji Burungale Science and Arts College
Shegaon Dist. Buldhana,-444 203, Maharashtra, India.
*Email: - [email protected]

ABSTRACT
The evaluation of density and viscometric study of 1-phenyl-3-[4-(2-ethylimino-4-t-
butylimino-1,3,5-dithiazino)aminophenyl]prop-2-ene-1-one was recently carried out in our
laboratory. In this research work evaluation done at different temperatures by keeping the
constant concentration. The experimental data determine the effect of dilution of the solvent
and the solute-solvent interaction of drug in current times.

KEYWORDS
Solute-solvent interaction, Viscometric study, 1,3,5-Dithiazino etc.

INTRODUCTION
The chalcones moiety base heterocyclic compounds are very widely distributed in
nature. The nitrogen and sulphur containing heterocyclic compounds are very essential to living
organisms. In biochemical, agricultural, pharmaceutical, medicinal, and industrial and drug
sciences (Solanki A. and Thakur I, 2007) (Saleem F, 2008) viscosity measurements play a
crucial role. Liquid have one of the important physical property is viscosity. Due to the shearing
effect in the liquid which is the movement of liquid layers over each other hence liquids are
viscous in nature (Bhat B.A., 2008). For aqueous and in non-aqueous solution have solute-
solute and solute-solvent interaction. This important information provided by measurements of
viscometric parameter. Drug behavior like absorption, transmission and its effect will directly
relate to its viscosity measurements and solvent interactions in the human framework (Vibhute
Y.B. and Basser M.A., 2008)

Literature review that chalcone derivatives exhibit diverse pharmacological and

biochemical activities such as antimicrobial and cytotoxic agents, antiviral, anti-inflammatory,
anestetics, mydriatics. Heterocyclic molecule having 1,3,5-dithiazino nucleus is widely used
in medicinal, biochemical, biotechnological and pharmaceutical sciences (Solanki A. and
Thakur I, 2007) (Saleem F, 2008) (Bhat B.A., 2008). These compounds showed anti-
helminthic, antifungal, antiviral, antibacterial and anti-tuberculostatic properties Vibhute Y.B.
and Basser M.A., 2008). Dithiazines are found to be effective on treatment of cancer (Wan
Z.Y., 2005). All these facts consideration a topic of great interest to carry out the viscometric
measurements of 1-phenyl-3-[4-(2-ethylimino-4-t-butylimino-1,3,5-
dithiazino)aminophenyl]prop-2-ene-1-one by varying temperatures (Jakhar A. and Makrand
J.K.,2010) Such kind of study will be helpful to drug effectiveness (Zhang L.X., Zhang A.J.,
2003) (Solanki A. and Thakur I, 2007) (Saleem F, 2008).

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EXPERIMENTAL
For all type of analysis double distilled water and A.R. grade chemicals were used
throughout the evaluation. For compound weight was used Mechaniki Zektady Precyzyjnej
Gdansk balance [Poland make (±0.001gm)]. For measurement of viscosity of liquid was used
Ostwald’s viscometer. It was kept in Elite themostatics water bath and temperature variation
was maintained at 290C (±0.1) for each measurement. For densities determination bicapillary
with a 1 mm internal diameter was used. Viscometer and water bath required sufficient time
for maintaining thermal equilibrium.
In this research work viscometric study of 1-phenyl-3-[4-(2-ethylimino-4-t-
butylimino-1,3,5-dithiazino)aminophenyl]prop-2-ene-1-one at 0.1M concentration in 60%
ethanol-water system separately at varying temperatures. Freshly prepared solutions were used
throughout the evaluation. As per the literature given viscometric readings were taken.

OBSERVATIONS AND CALCULATIONS

Molecular interactions in terms of β-coefficient of solute is figured with the help of data
obtained in our work. The results obtained are stated in Table No. 1. According to Jone’s-Dole
equation, (ηr-1)/√C = A+B√C at different temperatures keeping the concentration 0.1 M. A
and β-coefficient values calculated are enlisted in Table No.2.

TABLE No.1
VISCOSITY MEASUREMENTS AT CONSTANT CONCENTRATIONS AND
DETERMINATION OF RELATIVE AND SPECIFIC VISCOSITIES AT DIFFERENT
TEMPERATURES AT 0.1M
MEDIUM - 60% ETHANOL-WATER
Density (ήr-
Temp. Time
Conc. √C ρx103 ήr ήsp=ήr-1 1)/√C
(°C) (sec.) -3
(kg.cm ) (pa-s)
22 0.314 58 1.0913 0.069341 -0.930659 -2.9638
24 0.314 51 1.0893 0.067329 -0.932671 -2.9702
0.1 M
28 0.314 47 1.0676 0.05920 -0.9408 -2.9961
30 0.314 32 1.0565 0.06453 -0.93547 -2.9792

TABLE No.– 2
A and β Co-Efficient Values from Graphs for 60%.
FOR 1-PHENYL-3-[4-(2-ETHYLIMINO-4-t-BUTYLIMINO-1,3,5-
DITHIAZINO)AMINO- PHENYL]PROP-2-ENE-1-ONE
W-E
Temp° C Mean "A" β (Slope "m")
Mixture(%)
60 24 -2.9702 0.0073

RESULT AND DISCUSSION

The relative viscosity was determined by using following formula
ηr = Ds x ts / Dw x tw.
While Jone’s-Doles equation was used for the analysis of relative viscosities,
(ηr-1)/√C = A+B√C
Where,
A = Falkenhagen coefficient

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B = Jones-Dole coefficient
C = concentration of solutions
Solute-solute interaction was measures by using Falkenhagen coefficient (A) while solute-
solvent interaction was measures by using Jones-Dole coefficient (B).
The graph are plotted in between (ηr-1)/√C versus√C. The graph for each system gave linear
straight line gave value of β-coefficient.

CONCLUSIONS
In the current work monitor that the density and relative viscosity decreases with
increase in temperature. This is supported by the information that as the temperature increases
the solute-solvent interaction increases due to which solvation effect increases.
Pharmacodynamics and pharmacokinetics study of drug is useful investigation and informative
for society.

ACKNOWLEDGEMENT
I am very much thankful to Dr. D. T. Tayade, Professor, GVISH College, and Amravati
for kindly cooperation.

REFERENCES
1. Solanki, A. and Thakur, I. (2007). Indian Journal of Chemistry. 45(B): 517.
2. Saleem, F. (2008). Eur. Pot., CHAPPL. 87: 19.
3. Baldaniya, B.B. and Patel, P.K. (2009). E-Journal of Chemistry. 6(3): 673-680.
4. Parajuli, R. and Medhi, C. (2004). Journal of Chemical Sciences. 116(4): 235-241.
5. Vibhute, Y.B. and Basser, M.A. (2003). Ind. J. of Chem. 42(B): 202-205.
6. Bhat, B.A. Dhar, K.L. Saxena, A.K. and Shanmugavel, M. (2005). Bio org. and Med.
Chem. 15(3): 177-180.
7. Kalirajan, R. Palanivelu, M. Rajmanickam, V. Vinothapushan, G. and Anandarajagopal,
K. (2007). Int. J. of Chem. Sci. 5(1): 73-80.
8. Bansal, R.K. (2012). J.Heterocyclic Chloroic Chemistry. 8: 12-24.
9. Jakhar, A. and Makrand, J.K. (2010). J.Chem.Res. 4(3): 238-240.
10. Wan, Z.Y. Shi, H.X. and Shi, H.J. (2001). J. Heterocyclic Chloroic Chem. 38: 335.
11. Zhang, L.X. Zhang, A.J. Hu, M.L. and Lei, X.X. (2003). Acta Chim.Sinica. 61(6): 917.
12. Zhang, Y. Qiao, R.Z. and Zhang, Z.Y. (2002). J.Chin.Chem.Soc. 49(3): 369.

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67
An Overview on Green Chemistry
Dr. Rupali.S. Talegaonkar
Matoshree Vimalabai Deshmukh Mahavidyalaya, Amravati
E.Mail [email protected]

ABSTRACT
The present review work focuses on the importance and economic development of green
chemistry. It is new branch in chemistry dealing with reduction of harmful and toxic chemicals
in the synthesis and replacing it with ecofriendly methods. The principle of green chemistry
with various benefits have been discussed to understand the basic requirement for replacement
of conventional synthetic method with green chemistry synthesis. To describe it the synthetic
approach for the synthesis of acetanilide has been discussed and compared.

KEYWORDS: Green Chemistry, Ecofriendly, Conventional, Acetanilide.

1. INTRODUCTION
The accelerated progress in science and technology now a days has led to economic
development in world, but such economic development also cause environmental degradation
which is manifested by climate change, the issue of ozone holes and accumulation of non-
destructive organic pollutant in all parts of biospheres.
So the present situation required the solution to balance the use of natural resources and
environmental conservation. From last two decades awareness for environmental protection
has increased by using the concept of “Green Chemistry”. The new laws and regulations have
a Aim to protect the ecosystem from harmful chemicals and develop new compounds by the
approach of Green chemistry which are less dangerous to human health and the environment.[1].
Green chemistry is new branch of chemistry involves pulling together tools and
techniques that helps to chemical engineers in research related to the creation of chemical
product and processes that reduce or eliminate the use of harmful chemicals as well as reducing
harmful and toxic products for the development of more eco-friendly and efficient product with
less wastage. Green chemistry is now going to become an essential tool in the field of synthetic
chemistry [2,3].

1.1.Definition of Green Chemistry: According to environmental protection agency, green

chemistry is defined as a chemistry that designs chemical products and processes that are
harmless to the environment. Chemical products should be made in such a manner that they
do not remain in the environment at the end of their application and broken down into
components that are harmless to environment.4

1.2.History: The term green chemistry was first given by Poul .T. Anastas in 1991 in special
program launched by the US environmental Protection Agency (EPA) to implement
sustainable development in chemistry ,chemical technology by industry ,academia and
government. In 1995 the annual US presidential green chemistry challenge was
announced. In 1996 the working party on green chemistry was created, acting within the
framework of International Union of Pure and Applied Chemistry. The first book and
journals on the subject of green chemistry were introduced in 1990 by the royal society of
chemistry. Green chemistry includes a new approach to the synthesis, processing and

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application of chemical substances in such a manner to reduce scourge to health and

environment like:
 Clean chemistry
 Atom Economy
 Environment begin chemistry [5-11]
 Clean Chemistry
 Atom Economy
Twelve principles of Green chemistry have been developed by Poul Anastas, speaks about the
reduction of dangerous or harmful substances from the synthesis, production and
application of chemical products. When designing a green chemistry process it is impossible
to meet the requirements of all twelve principles of the process at the same time, but it
attempts to apply as many principles during certain stages of synthesis.[12,13]

1.3. Principles of Green Chemistry

1.3.1. Pollution Prevention: It is to prevent waste and to treat and clean up waste after it has
been created.
1.3.2. Atom Economy: It should be designed to maximize the incorporation of all materials
used in process into the final product.
1.3.3. Less Hazardous Chemical Synthesis: The synthetic method should be designed to use
and generate substances that process little or no toxicity to human health and the
environment.
1.3.4. Designing Safer Chemicals: Chemical product should be designed to affect their
desired function while minimizing toxicity.
1.3.5. Safer Solvents and Auxiliaries: The use of auxiliary substances should be made
unnecessary wherever possible.
1.3.6. Design for Energy Efficiency: energy requirements of chemical processes should be
recognized at low temperature and pressure.
1.3.7. Use of Renewable Feedstock’s: A raw material of feedstock should be renewable.
1.3.8. Reduce Derivatives: The unnecessary derivatization like use of blocking groups,
protection, deprotection should be avoided whenever impossible.
1.3.9. Catalysis: The catalytic reagents are superior stoichiometric reagents.
1.3.10. Design for Degradation: The chemical products should be designed so that at the
end of their function they breakdown into harmless degradation products and do not persist in
the environment.
1.3.11. Real-time analysis for Pollution Prevention: Analytical methodologies need to be
further developed to allow for real-time, in process monitoring and control prior to
the formation of hazardous substances.
1.3.12. Inherently Safer Chemistry for Accident Prevent: The substances used in the
chemical process should be chosen to minimize the potential for chemical accidents,
explosion and fires. This principle can motivate chemistry at all levels like research,
education and public perception. [14]

1.4. Benefits of Green Chemistry

Benefits for health
 Cleaner air- less release of hazardous chemicals to air leading to less damage
to lungs.
 Cleaner water- less release of hazardous chemicals waste to water leading to
cleaner drinking.
 Increase safety water for workers in chemical industry, less use of toxic
material, less potential for accident

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 Benefits for environment

 Plants and animals suffer less harm from toxic chemicals in environment
 Lower potential for Global warming, ozone depletion and smog formation.
 Economy
 Better performance so that less product is needed to achieve the same
function.
REVIEW LITERATURE
2.1. Green chemistry aims to reduce or even eliminates the production of any harmful bi-
products and maximizing the desired product without compromising with the environment.
The three key developments in green chemistry include use of super critical carbon di oxide as
green solvent, aqueous hydrogen peroxide as an oxidizing agent and use of hydrogen in
asymmetric synthesis. It also focuses on replacing traditional methods of heating with that of
modern methods of heating like microwave radiations so that carbon footprint should be
reduces as low as possible.[15]
2.2. The work focuses to reduce the chemical wastage by applying the concept of green
chemistry. Few derivative of acetanilide were synthesized by conventional method as well
as by green chemistry method. In conventional method there was wastage of chemicals
by the formation of acetic acid molecule but by green synthesis method the formation of
byproducts was avoided and the atom economy was calculated on the basis of molecular weight
of desired product and it was found to be in the range of 72 to 82% which signify the utility of
green synthesis method.[16]
2.3. Due to technology development the quality of life on earth became much better but
harmful effect of chemistry also became pronounced main among them being the
pollution of land, water and atmosphere. This is caused mainly due to the use of harmful
reactants and effect of by-product of chemical industries, which are being discharge into air,
rivers and the land, but by applying the concept of green chemistry these all problems can be
reduced.[17]
2.4. Green chemistry is a term that refers to the production of chemical products and
processes that reduce the use of and production of harmful substances[18]
The green chemistry revolution provides the various numbers of challenges to those who
practice chemistry in industry, education and research. It is the modern science of
chemistry deals with the application of environment friendly chemical compound in the
various area of our life such as industries. The chemical industries supply us a huge variety
of essential product, from plastic to pharmaceuticals, these industries has a potential to
damage our environment, so green chemistry serves to promote the design and efficient use of
chemicals and chemical processes [19]

3. EXPERIMENTATION
The synthesis of acetanilide by green chemistry has various disadvantages as there is a lot of
wastage of acetic acid molecule, which can be minimized by green chemistry.

3.1. Synthesis of compounds by conventional method

 In a 250 ml beaker containing 125 ml of water, 4.6 ml of conc. Hydrochloric acid and
5.1
 In a 250 ml beaker containing 125 ml of water, 4.6 ml of conc. Hydrochloric acid and
5.1 g of aniline / substituted anilines were introduced.
 Stirred until all the anilines passes completely into solution.
 To the resulting solution, 6.9 g (6.4ml) of redistilled acetic anhydride was added and
stirred until it was dissolved.

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 Poured immediately in a solution of 3.8 g of crystalline sodium acetate in 25 ml of

water.
 Stirred vigorously and cooled in ice. Filtered the acetanilide and substituted
acetanilide with suction, washed with 10 ml water, drained well and dried upon filter
paper. The crude products were recrystallized from boiling water and methylated
spirit.
 Non green component: Acetic anhydride leaves one molecule of acetic acid unused.
 Poured immediately in a solution of 3.8 g of crystalline sodium acetate in 25 ml of
water.
 Stirred vigorously and cooled in ice. Filtered the acetanilide and substituted
acetanilide

3.2. Synthesis of compounds by green chemistry method

 A mixture of aniline / substituted anilines (3.3g) and zinc dust (0.16g) in acetic acid
 A mixture of aniline / substituted anilines (3.3g) and zinc dust (0.16g) in acetic acid (10ml) in
100 ml round bottom flask was heated over a gentle flame using water condenser.
 Heating was continued for about 45 min., the reaction mixture was then carefully poured in
cold water (33ml) in 250ml beaker with vigorous stirring.
 The shining crystals of product were separated slowly. After 15 min, crystals were collected
by filtration. The solid crystals were washed over the Buchner funnel with water and product
was dried and crystallized in boiling water.
 Green context: Minimize waste by-products, avoids use of acetic anhydride
 Benefits for health
 Cleaner air-less release of hazardous chemicals to air leading to less damage to lungs.
 Cleaner water-less release of hazardous chemical wastes to water leading to cleaner

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4. CONCLUSION
This review article will definitely help to understand the importance of green chemistry,
conventional methods can be replaced easily by the methods which utilize non toxic and
environment friendly techniques for the synthesis of same. One of such approach has been
discussed for the synthesis of acetanilide. The approach will definitely help in the synthesis by
keeping the environment safe. which is basic requirement in today’s pharmaceutical
industries. The approach will help to avoid the utilization of the toxic chemicals leading to
various hazards in the industry. These

5. REFERENCE
1. Singhal M., Singh A. Khan S.P, Green Chemistry Potential for Past, Present and Future Perspectives,
2012; 3(4).
2. Ahuwalia V.K, Kidvai M., New Trends In Green Chemistry,Anamayapublisher New Delhi, 2nd
edition, 2007; 5-18.
3. Ahluwalia V.K, Green chemistry Enviromentally Benign Reactions, published by India books, 2 nd
EDITION, 2006; 1-10.
4. Vojvodic V. Enviromental Protection :Green Manufacturing in the pharmaceutical industry and cost
reduction, KenInd, 2009; 58(1): 32-33.
5. Anastas. P.T, Warner J.C, Green chemistry Theory and Practice, Oxford University, Press, New York,
1998.
6. Anastas P.T, Hovarsth I.T, Innovations and Green Chemistry, Chemistry review, 2007; 107.
7. Ravichandaran S., International Journal, 2010; 2(4): 2191.
8. Trost B.M, Atom economy- A challenge for organic synthesis:Hompgeneous catalysis leads the way,
1995; 34: 259.
9. Sheldon R.A, Green solvents for sustainable organic synthesis: State of art, 2005; 7: 267.
10. Bharati V.B, Resonance, 2008; 1041.
11. Ahluwalia V.K and Kidwai M., New Trends in Green Chemistry, Anamaya Publisher, New Delhi,
2004.
12. Anastas P., Warner, Green Chemistry: Theory and Pracxtice, Oxford University Press, Oxford, 1998.
13. Anastas P.T, Heine L.G, Williamson T.V, Green Chemical Synthesis and Processes, American
Chemical Soceity, Washington DC, 2000.
14. Singhal M, Singh A, Khan S.P, Sultan E, Sachan N.K, Green chemistry potential for past present and
future perspectives.
15. Gujral. S.S, Sheela. M.A, Khattri S., Singhla R.K. A Focus and Review on the Advancement of
Green Chemsitry, Indo Global Journal of Pharmaceutical Science, 2012; 2(4): 397-408.
16. Redasani V.K, Kumawat V.S, Kabra R.P, Surana S.J, Application Of Green Chemistry in Organic
Synthesis, International Journal of Chem Tech Research, 2010.
17. Singhal. M, Singh A., Khan S.P, Green Chemistry Potential for Past, Present and Future Perspectives,
International Research Journal of Pharmacy, 2012; 3(4).
18. Ivankovic. A., Dronjic A., Review of 12 Principles of Green Chemistry in Practice, International
Journal of Sustainable and Green Energy, 2017; 6(3): 39-48.
19. Chanshetti U., Green Chemistry: Challenges And Opportunities In Sustainable Development,
International Journal of Current Research, 2014; 6.

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68

3D Printing Technology for development of Transdermal Drug Delivery

Systems

Dr. Amar Deshpande, Dr. Jagdish Baheti

Kamla Nehru College of Pharmacy, Butibori, Nagpur

Abstract:
The innovative approach of three-dimensional printing enables the on-demand production of
transdermal drug delivery systems. This technology, already applied in dentistry, orthopedics,
and pharmaceuticals, particularly stands out in the latter field. It facilitates the printing of
medical devices and diverse formulations of active pharmaceutical ingredients, featuring
controlled-release characteristics and varied geometries. This study provides an overview of
these pharmaceutical applications, focusing on the 3D printing of transdermal patches. The
discussion encompasses different printing technologies and material systems known for their
customization capabilities, generating intricate geometries with precise characteristics crucial
for transdermal systems, thereby enhancing bioavailability. The study includes case studies,
explores advantages and limitations of the technology, and forecasts industry growth,
projecting a value exceeding USD 8 Billion by 2025. Despite this potential, the conservative
nature of the pharmaceutical industry leans toward cost-effective methods for large-scale
production. Nevertheless, 3D printing has the potential to revolutionize the current 'one size
fits all' manufacturing approach, becoming an integral part of the drug development timeline.

Keywords: 3D Printing, Inkjet printing, Microneedles, Patches, Transdermal delivery,

Pharmaceutics

1. Introduction
The historical application of therapeutic substances, including herbal ointments and various
drugs (such as scopolamine, estradiol, fentanyl, rivastigmine), on the human skin serves both
medical and cosmetic purposes. Over the past decades, the skin has proven to be an accessible
surface for drug administration, making systematic therapy through percutaneous drug
absorption feasible (Prausnitz and Langer, 2008; Alkilani et al., 2015; Pastore et al., 2015). The
transdermal route emerges as an attractive alternative to traditional methods like oral
administration or hypodermic injections. Oral administration may face issues of partial drug
absorption, complications related to gastrointestinal metabolism, and slow onset, making it
impractical for emergency cases. Hypodermic injections, while effective, are invasive, pose
infection risks, require skilled administration, and generate medical waste (Awodele et al.,
2016).
Contrarily, transdermal systems offer advantages such as bypassing metabolic systems,
ensuring higher bioavailability, and promoting sustained and controlled drug release.
Additionally, transdermal drug delivery (TDD) holds promise for vaccinations due to the
abundance of dendritic cells in the skin. This patient-friendly approach is noninvasive,
contributing to psychological well-being, and provides independence as it doesn't require
professional care for repositioning, removal, or replacement.
However, TDD faces limitations, primarily stemming from the skin barrier's nature. The
Stratum Corneum, the outermost skin layer, acts as a significant barrier due to its density and
low hydration (15–20%). Overcoming this impermeable barrier has been the focus of TDD
research, presenting both challenges and opportunities for future progress.

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In the current global trend towards personalized patient care, traditional mass-production
methods of drug delivery systems are being questioned for their ability to tailor dosages cost-
effectively. New technologies, particularly Additive Manufacturing (AM), such as 3D printing,
are investigated for their potential in pharmaceutical technology. Initially introduced in the
1980s, 3D printing has gained attention across various industries, contributing to the production
of complex structures beyond the capabilities of conventional techniques.
The application of 3D printing in pharmaceuticals is relatively recent, aiming to produce
targeted-release and customized drug delivery systems (Goole and Amighi, 2016). In the field
of TDD, although studies are limited, they demonstrate the transformative potential of 3D
printing. This review explores the existing research on 2D and 3D printing as direct or indirect
fabrication methods for TDD systems. The materials and drugs associated with 3D printing in
TDD systems are also examined in this context.

2. 3D printing techniques for optimized Transdermal Drug Delivery Systems

Additive Manufacturing (AM), commonly referred to as 3D printing or Solid Freeform
Fabrication (SFF), encompasses various techniques that utilize a virtual Computer Aided
Design (CAD) model to construct a physical object by depositing consecutive layers.
Introduced in the 1980s, 3D printing has revolutionized industrial and scientific sectors,
offering fast and precise production of intricate structures beyond the capabilities of traditional
methods. The medical field quickly recognized the transformative potential of 3D printing,
leading to the creation of customized implants, prosthetics, and ongoing investigations into live
tissue printing (Chia and Wu, 2015).
The application of 3D printing in drug delivery has recently gained attention, with the FDA
approval of Spritam, the first 3D printed oral administration tablet. This has given rise to the
term 'pharmacoprinting' (Prasad and Smyth, 2015; Jacob et al., 2014; Goyanes et al., 2015; Di
Prima et al., 2016). While its impact on oral drug delivery is well-established, 3D printing's
potential in transdermal drug delivery (TDD) is currently under exploration, with a growing
body of relevant studies.

2.1. Inkjet Printing

Inkjet printing, involving the controlled deposition of small droplets, has seen successful
applications in medicine but is yet to be extensively explored for TDD. Studies have used inkjet
printing for coating microneedles with various agents, demonstrating its potential for controlled
and selective deposition on suitable substrates (Boehm et al., 2011, 2013, 2014; Ross et al.,
2015; Uddin et al., 2015). While its application for building complex three-dimensional TDD
structures remains unexplored, inkjet printing's high resolution and selective deposition make
it promising for microneedle coating, enabling personalized dosages with high reproducibility.

2.2. Photopolymerization-based Technologies

A significant group of 3D printing technologies relies on selective polymerization of photo-
sensitive polymers through laser emissions or light projections. Techniques like
Stereolithography (SLA) and Digital Light Processing (DLP) enable layer-wise polymerization
of UV-sensitive polymers. These technologies offer versatility in geometric complexity and
resolution, making them suitable for TDD applications. Studies have utilized micro-
stereolithography (DLP) to create microneedle arrays indirectly, contributing to the
customization of therapeutic approaches (Boehm et al., 2014, 2011, 2012).
Photopolymerization-based 3D printing has proven applications in fabricating TDD systems,
offering high resolution and flexibility.

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2.3. Fused Deposition Modelling (FDM)

Fused Deposition Modelling (FDM), based on the melt-extrusion process, is a versatile 3D
printing technique with potential applications in TDD. While FDM's limitations in resolution
and sensitivity to process parameters are acknowledged, its ability to produce structures
through extrusion without solvents makes it a compelling choice for certain pharmaceutical
applications. The combination of FDM with hot-melt extrusion (HME) processes holds
promise for producing drug/polymer blends for cost-effective TDD system fabrication.

3. Materials:
In the contemporary landscape, 3D printing technologies possess the capability to manipulate
a diverse array of materials, ranging from ceramics and metals to polymers. The categorization
of these techniques implies that the choice of a specific technology inherently limits the
materials compatible with the corresponding printing apparatus. For instance, SLA or DLP
printers exclusively handle photo-cured polymers, while FDM printers utilize thermoplastic
filaments. This limitation poses challenges for the widespread adoption of 3D printing as a
direct manufacturing technique for Transdermal Drug Delivery (TDD) systems, as the material
must meet specific criteria for integration into such systems. (Sharma et al.2011) Essential
parameters include stability, biodegradability without toxic by-products, mechanical strength,
and non-reactivity with the drug. Material biocompatibility is a critical consideration, as
evidenced by studies involving Gantrez, a biocompatible copolymer used in TDD applications.
While there's evidence of manufacturing Gantrez biocompatible microneedles using 3D printed
molds, the multi-step nature of this approach may hinder mass production scalability. (Boehm
et al., 2014, 2011, 2012, Donnelly et al., 2012).
Numerous polymers with biomedical and pharmaceutical applications show promise for
integration into 3D printed TDD systems. Polyvinyl alcohol (PVA) and poly lactic acid (PLA)
are examples. However, challenges exist, such as PVA's limited biodegradability and PLA's
slow degradation rates and poor mechanical properties when employed in FDM technology.
Biopolymers, like chitosan and collagen, exhibit favorable attributes for TDD. Bioprinting
advancements further expand the possibilities. Yet, ongoing research on materials remains vital
for the evolving field of 3D printed TDD. (Economidou et al. 2018)

4. 3D Printed Transdermal Drug Delivery Systems- Future Challenges and Expected

Impact:
Transdermal Drug Delivery (TDD) systems, facilitated by 3D printing, hold potential as a user-
friendly, personalized pharmaceutical therapy. The layer-by-layer fabrication inherent in 3D
printing aligns well with TDD requirements. This technology enables the creation of systems
with varying drug concentrations across layers, catering to individual needs. Customization
possibilities enhance TDD efficiency, addressing factors like skin hydration and thickness
variations among patients. In vaccination, microneedles offer promise, particularly in regions
facing challenges with traditional administration methods. 3D printing's role in reducing costs
and providing needle-free solutions is crucial for global health initiatives. However, challenges
such as limited biomaterial options, dosing constraints, and drug degradation characteristics
need resolution. The development of 3D printable materials and improvements in existing
technologies could drive the evolution of TDD systems. (Economidou et al. 2018)

5. Regulatory Considerations:
For the commercialization of 3D printed TDD systems, adherence to regulatory requirements
is essential. Despite the FDA's approval of the first 3D printed oral tablet, TDD systems face
unique regulatory challenges. Microneedle patches, viewed as medical devices, must adhere to
Good Manufacturing Practice (cGMP) guidelines. The FDA emphasizes technical

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considerations, including the impact of printing parameters, in-situ quality control, design
validation, sterilization, and post-process cleaning. A 2017 guidance document provides
recommendations for 3D printed medical devices, addressing issues like patient-matched
devices and data protection. Sterilization, a regulatory requirement, presents challenges for
microneedles. Future success depends on addressing material limitations, improving
technology, and establishing specific regulatory frameworks for 3D printed TDD systems.
(Economidou et al. 2018)

6. Conclusion:
Since its inception, 3D printing has revolutionized fabrication methods, with potential
applications in medicine and pharmaceutics. Despite being in the early stages, advancements
in 3D printed Transdermal Drug Delivery (TDD) systems show promise. Inkjet printing,
photopolymerization-based technologies, and FDM have been explored, with inkjet printing
successfully depositing films on microneedle surfaces and commercializing 3D inkjet printed
microneedles. The integration of elaborate microneedle array systems with precise 3D printing
techniques has the potential to reshape modern drug administration. Overcoming engineering,
chemistry, and material challenges through interdisciplinary research is crucial. Regulatory
considerations, addressed by the FDA, are imperative to ensure the safety and effectiveness of
3D printed TDD systems. Success hinges on resolving material limitations, advancing
technology, and establishing specific regulatory frameworks.

References:
1. Alkilani, A.Z., McCrudden, M.T.C., Donnelly, R.F., 2015. Transdermal drug delivery:
innovative pharmaceutical developments based on disruption of the barrier properties
of the stratum corneum. Pharmaceutics 7 (4), 438–470.
2. Awodele, O., Adewoye, A.A., Oparah, A.C., 2016. Assessment of medical waste
management in seven hospitals in Lagos, Nigeria. BMC Public Health 1–11.
3. Boehm, R.D., Miller, P.R., Hayes, S.L., Monteiro-Riviere, N.A., Narayan, R.J., 2011.
Modification of microneedles using inkjet printing. AIP Adv. 1 (2), 022139.
4. Boehm, R.D., Miller, P.R., Singh, R., Shah, A., Stafslien, S., Daniels, J., Narayan, R.J.,
2012. Indirect rapid prototyping of antibacterial acid anhydride copolymer
microneedles. Biofabrication 4 (1), 11002.
5. Boehm, R.D., Miller, P.R., Schell, W.A., Perfect, J.R., Narayan, R.J., 2013. Inkjet
printing of amphotericin B onto biodegradable microneedles using piezoelectric inkjet
printing. JOM 65 (4), 525–533.
6. Boehm, R.D., Miller, P.R., Daniels, J., Stafslien, S., Narayan, R.J., 2014. Inkjet printing
for pharmaceutical applications. Mater. Today 17 (5), 247–252.
7. Chia, H.N., Wu, B.M., 2015. Recent advances in 3D printing of biomaterials. J. Biol.
Eng. 9 (4).
8. Di Prima, M., Coburn, J., Hwang, D., Kelly, J., Khairuzzaman, A., Ricles, L., 2016.
Additively manufactured medical products – the FDA perspective. 3D Print. Med. 2
(1), 1–6.
9. Donnelly, R.F., Singh, T.R.R., Garland, M.J., Migalska, K., Majithiya, R., McCrudden,
C.M., Kole, P.L., Mahmood, T.M.T., McCarthy, H.O., Woolfson, A.D., 2012.
Hydrogel forming microneedle arrays for enhanced transdermal drug delivery. Adv.
Funct. Mater. 22 (23), 4879–4890.
10. Economidou, S. N., Lamprou, D. A., & Douroumis, D., 2018. 3D printing applications
for transdermal drug delivery. International journal of pharmaceutics. 544(2), 415-424.

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11. Gittard, S.D., Miller, P.R., Jin, C., Martin, T.N., Boehm, R.D., Chisholm, B.J.,
Stafslien, S.J., Daniels, J.W., Cilz, N., Monteiro-Riviere, N.A., Nasir, A., Narayan, R.J.,
2011.
12. Goole, J., Amighi, K., 2016. 3D printing in pharmaceutics: A new tool for designing
customized drug delivery systems. Int. J. Pharm. 499 (1–2), 376–394.
13. Goyanes, A., Wang, J., Buanz, A., Martinez-Pacheco, R., Telford, R., Gaisford, S.,
Basit, A.W., 2015. 3D printing of medicines: engineering novel oral devices with
unique design and drug release characteristics. Mol. Pharm. 12 (11), 4077–4084.
14. Jacob, J., Coyle, N., West, T.G., Monkhouse, T.G., Surprenant, H.L., Jain, M.B., 2014.
Rapid Disperse Dosage form Containing Levetiracetam. US 20140271862 A1.
15. Jessen, L., Kovalick, L.J., Azzaro, A.J., 2008. The selegiline transdermal system
(Emsam): a therapeutic option for the treatment of major depressive disorder. Pharm.
Ther. 33 (4), 212–246.
16. Pastore, M.N., Kalia, Y.N., Horstmann, M., Roberts, M.S., 2015. Transdermal patches:
history, development and pharmacology. Br. J. Pharmacol. 172 (9), 2179–2209.
17. Prasad, L.K., Smyth, H., 2015. 3D printing technologies for drug delivery: a review.
Drug Dev. Ind. Pharm. 9045 (January), 1–13.
18. Prausnitz, M.R., Langer, R., 2008. Transdermal drug delivery. Nat. Biotechnol. 26 (11),
1261–1268.
19. Ross, S., Scoutaris, N., Lamprou, D., Mallinson, D., Douroumis, D., 2015. Inkjet
printing of insulin microneedles for transdermal delivery. Drug Deliv. Transl. Res. 5
(4), 451–461.
20. Sharma, K., Singh, V., Arora, A., 2011. Natural biodegradable polymers as matrices in
transdermal drug delivery. Int. J. Drug Dev. Res. 3 (2), 85–103.
21. Uddin, M.J., Scoutaris, N., Klepetsanis, P., Chowdhry, B., Prausnitz, M.R., Douroumis,
D., 2015. Inkjet printing of transdermal microneedles for the delivery of anticancer
agents. Int. J. Pharm. 494 (2), 593–602.

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69

In-silico Prediction of Phytoconstituents from Hymenodictyon

excelsum for Antimalarial Activity Targeting Hypoxanthine-guanine
phosphoribosyltransferase (HPRT1)
Parimal Katolkar*, Jagdish Baheti
Department of Pharmaceutical Chemistry, Kamla Nehru College of Pharmacy, Butibori, Nagpur 441108 (MS),
INDIA
*Email: [email protected]

ABSTRACT
Objective Malaria is referred as a disease cause by parasite. The parasite spread to humans
through the infected mosquito bite. People who infected by malaria feel very sick with high
fever and chills. Natural products and their active principles as sources for new drug discovery
and treatment of diseases have attracted considerable attention of researchers. Compounds
found in medicinal plants have been the source of many conventional medications. In-silico
testing of Hymenodictyon excelsum phytoconstituents on hypoxanthine-guanine
phosphoribosylt ransferase for antimalarial efficacy was a part of our investigation.

Methods Utilizing Discovery studio, molecular docking is done to assess the pattern of
interaction between the phytoconstituents from the Hymenodictyon excelsum plant and the
crystal structure of the malarial proteins (PDB ID: 4RAC). Later, SwissADME and pkCSM
were used to screen for toxicity as well as the pharmacokinetic profile. Further, interaction
between proteins by STRING network analysis and bioactivity score analysis using
molinspiration tool were also evaluated.

Results The docked results suggest that nordamnacanthal (−8.9 kcal/mol), rubiadin (−8.2
kcal/mol) and lucidin (− 8.1 kcal/mol) for 4RAC macromolecule has best binding towards
antimalarial activity as compared to the standard (artemisinin) for 4RAC is -7.5 kcal/mol and.
Furthermore, pharmacokinetics and toxicity parameters were within acceptable limits
according to ADMET studies.

Conclusions Targeting hypoxanthine-guanine phosphoribosyl transferase against malaria by

phytoconstituents from Hymenodictyon excelsum can serveas a rational approach for designing
future antimalarial drugs.

1. INTRODUCTION
About 30 species of flowering plants belong to the genus Hymenodictyon, which is part of the
Rubiaceae family. The generic name is a combination of the Greek words for membrane and
net, hymen and diktyon. It alludes to the wing that envelops every seed. The members in this
genus have serrated leaves grouped in opposition, small, clustered blooms, and many seeds
capsules. The genus includes widely varying trees and shrubs the majority of them in tropical
and subtropical regions of Asia and Africa.1
Hymenodictyon excelsum Roxb. Wall. syn H. orixense Roxb Mobb. belonging to family
Rubiaceae, also known as Bhorsal, Kukurkat, Bhaulan, Bauranga, Pottaka, Kusan, and
Kadambu (India), Lala (Thailand), and Kuthan (Burma) is a medium to a tall, deciduous tree
that typically grows to a height of 10 to 12 metres grey-brown with hints of brown, straight
cylindrical bole, rounded crown green, glabrous leaves that are rectangular, ovate, or elliptic in
shape, greenish-white, fragrant flowers, and ellipsoid capsules that contain flying seeds Bark

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is 10-20 cm long and mostly ruffled, thick, softish scales that exfoliate in irregular shapes.
Stipules are approximately 15 mm length, linear to lanceolate to ovate to lanceolate, apex
pubescent, acuminate, and deciduous.2-4
In traditional medical systems all across the world, H. excelsum's bark and leaves are primarily
used to cure a variety of illnesses. Bark has a variety of medical benefits. Different plant
components have reportedly been used to treat emaciation, a carbuncle, a burning sensation in
the chest, as well as problems related to lactation and fever. Additionally, it makes you taste
and eat more. Inhalation remedy for diarrhoea.5-6 It has significant in-vitro and in-vivo
antimalarial action since the ethyl acetate extract of H. excelsum has showed dose-dependent
percentage suppression of Plasmodium falciparum schizont formation.7
Coumarins8 and anthraquinones9, two chemical components previously identified as being
present in this plant, were detected. The stem bark of the plant contains tannin, the poisonous
alkaloid hymenodictine, aesculin, an apioglucoside of scopoletin, and hymexelsin.6,10
Additionally isolated from roots are anthraquinones, rubiadin and its methyl ether, lucidin,
damnacanthal, nordamnacanthal, 2-benzylzanthopurpurin, anthragallol, soranjidol, and
Morindone.4 Anthragallol, 6-methyalizarin, subiadin, and its 1-methylester, soranjidol, isolated
from roots; aesculin (b-methylaesinietin), scopoletin, and hymenodictyonim; alanine, arginine,
cystine, glycine, leucine, fruitore, galactose, and glucose,11acetylenic fatty acids, a novel
triglyceride, and 11 recognised chemicals, including ursolic acid, oleaqnolic acid, uncarinic
acid E, and β-sitosterol, have also been reported in studies.8
Malaria is the most common infectious disease in Africa and many Asian nations. Malaria is
caused by a microscopic protozoon from the Plasmodium species family, which comprises
numerous subspecies. A few Plasmodium species can cause illness.12,13
Five of the 172 Plasmodium species that exist can infect people. These include P. knowlesi, P.
vivax, P. ovale, P. vivax, P. malariae, and P. falciparum.13-16 Malaria is a disease that is caused
by all of the aforementioned Plasmodium species.17
Female Anopheles mosquitos are the only ones that transmit human malaria. After being bitten
by an infected female mosquito, the parasite enters the human blood as a sporozoite and travels
on to the hepatocytes after 30 minutes of blood circulation.18

4RAC Macromolecule: As the only pathway for the synthesis of the purine nucleoside
monophosphates necessary for DNA/RNA creation, hypoxanthine, guanine, and [xanthine]
phosphoribosyltransferase (HG[X]PRT) is regarded as a crucial target for antimalarial
chemotherapy.
In many species, the enzyme hypoxanthine-guanine-[xanthine] phosphoribosyl transferase
(HG[X]PRT) plays a vital role. It causes a purine base and a 5′-phospho-D-ribosyl-1-
pyrophosphate to combine to generate 6-oxopurine nucleoside monophosphates (PRib-PP).19
Therefore, replication should stop if this enzyme is inhibited. The Plasmodium falciparum
HGXPRT is effectively inhibited by the aza-acyclic nucleoside phosphonates (aza-ANPs) (Pf
HGXPRT).
Although it has been hypothesised for a long time that Plasmodium falciparum HGXPRT
(PfHGXPRT) could be a target for medications intended to treat malaria20, inhibitors have only
recently been transformed into substances with antimalarial action.21-22
However, there are few studies on the phytoconstituents of Hymenodictyon excelsum for the
antimalarial activity. Thus, keeping the above information in view, the present investigation
was designed to identify the potential phytochemicals of Hymenodictyon excelsum against
4RAC using a molecular docking method.

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2. MATERIALS AND METHODS

2.1. Platform for molecular docking
The computational docking study of all the phytoconstituents selected as ligands with
antimalarial activity as the target was performed using PyRx software.23
2.2. Protein preparation
The macromolecule is 4RAC, in-silico analysis of selected phytoconstituents was performed
on the 2.05 Å crystal structure of antimalarial macromolecule with inhibitor, (PDB ID:4RAC
having resolution: 2.05 Å, R-Value Free: 0.244, R-Value Work: 0.194, R-Value Observed:
0.196), which was retrieved from the protein data bank (https://www.rcsb.org) 4RAC is
classified as Tranferase/Transferase inhibitors. all other molecules, such as co-crystallized
water molecules, unwanted chains, and nonstandard residues, were deleted. Using Discovery
studio.24

2.3. Ligand preparation

The three-dimensional (3D) structures of all constituents were retrieved using Avogadro
software from the PubChem database available on the NCBI website
(https://pubchem.ncbi.nlm.nih.gov/). However, the drawing of geometrical 2D structure was
performed using the ChemSketch program. The two-dimensional (2D) structures were
transformed into 3D models using the Avogadro software and the ligand structures were saved
in the PDB format. All the chemical structures are shown in Figure 1.

Rubiadin Lucidin
OH O OH
O
CH3 OH

OH
OH
O O

Damnacanthal Nordamnacanthal
-
O O O OH

O
O

OH OH

O O

2-benzylzanthopurpurin Morindone
O OH
O
HO OH

H3C
HO HO O
O

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Ursolic acid
CH3 Aesculin
OH
H3C
HO O

OH HO
CH3 CH3 H
O
O
H
HO O O
HO
H3C H
CH3

Scopoletin
H3C O

HO O O

Fig. 1. Chemical structures of all selected phytoconstituents in the molecular docking

studies
2.4 Standard Preparation
The standard is prepared steps such as, the 2D structure of standard drug was made using chem
sketch program, then the 2D structure was converted into 3D model using Avogadro Software,
it was saved in PDB format.
By using PyRx molecular docking of Artemisinin was done with 4RAC.

2.5. Molecular docking

Molecular docking evaluates the protein-ligand interactions and estimates the scoring function
based on the geometry to predict the binding affinity of the ligand molecule 25-26. We
applied molecular docking studies to investigate the binding pattern of selected
phytoconstituents (Figure 1) and the standard drug, along with the crystal structure of
antimalarial activity macromolecule (PDB ID:4RAC). The molecular docking study was
performed using PyRxsoftware, Binding affinity was explored using the Vina wizard tool. The
final results were analysed and visualized using Discovery Studio 2020 Client 27, with bound
ligands as the standard. Visualization of protein ligand interaction reflects the number of
interactions and active residues responsible for significant binding at the active site of the target
enzyme.

2.6. Absorption, distribution, metabolism, and excretion (ADME) and toxicity

prediction
The selected phytoconstituents and standard drug were further checked for drug-likeness
properties according to Lipinski’s rule. During drug development, it is necessary to predict the
tolerability of phytochemicals before being ingested by humans and animal models.
The pharmacokinetic profile (ADME) and toxicity predictions of ligands were conducted using
SwissADME (http://www.swissadme.ch) and pkCSM (an online server database predicting
small-molecule pharmacokinetic properties using graph-based
signatures, (http://biosig.unimelb.edu.au/pkcsm/prediction). To analyse the toxicological
properties of ligands, Simplified Molecular Input Line Entry System (SMILES) notations or

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PDB files were uploaded, followed by selecting the required models for generating numerous
information about structure-related effects28-29.

2.7 Interaction between Proteins by STRING Network Analysis

Search Tool for Retrieval of Interacting Genes (STRING) (https://string-db.org/) Database
predicts and integrates protein–protein interaction to identify functional relationships and
interactions between proteins. To seek potential interactions between genes involved in DNA
Polymerase λ, the STRING tool was employed. STRING provided a platform that postulated
several nodes, edges, average node degrees, protein–protein interaction (PPI) enrichment p-
values, and average and local clustering coefficient. STRING provided possible biological
processes, molecular function, and cellular components of candidate genes studied. 30

2.8 Bioactivity Score Analysis Using Molinspiration Tool

Molinspiration (http://www.molinspiration.com/cgi-bin/properties) predicts the drug resemblance
properties of the compound dependent on various descriptors. Drugs entering the body should
tie to an organic molecule to communicate its movement. Along their path, the bioactivity of
compounds was anticipated by utilizing the Molinspiration tool which gave a bioactivity score
of the phytocompound against the human receptors like GPCRs, ionic channels, kinases,
various receptors, proteases, and proteins. A complex is considered to be dynamic if the
bioactivity score is more than 0.0, modestly dynamic if somewhere in the range of−5.0 and 0.0,
and idle if under−5.0. 31

3. RESULTS AND DISCUSSION

The present study aimed to explore the inhibitory potential of the phytoconstituents present
in Hymenodictyonexcelsum targeting antimalarial activity. In this study, we performed
molecular docking studies of all phytoconstituents found
in Hymenodictyonexcelsumusing AutoDock Vina, followed by a study of interacting amino
acid residues and their influence on the inhibitory potentials of the active constituents. Selected
phytoconstituents showing the best fit were further evaluated for absorption, distribution,
metabolism, excretion, and toxicological (ADMET) properties using SwissADME and pkCSM
servers.

3.1 Molecular docking

The docking scores and binding energies of all chemical constituents
of Hymenodictyonexcelsumtargeting antimalarial activity (PDB ID:4RAC) and binding
interactions with amino acid residues are presented in Table 1 respectively.

Table 1. Binding interaction of ligands from Hymenodictyon excelsum

targeting antimalarial activity (PDB ID:4RAC)

Docking Score
Sr. No. Chemical constituent PubChem ID
4RAC
1. Rubiadin 124062 -8.2
2. Lucidin 10163 -8.1
3. Damnacanthal 2948 -8.4
4. Nordamnacanthal 160712 -8.9

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5. 2-benzylzanthopurpurin 630215 -7.8

6. Morindone 442756 -8.2
7. Ursolic Acid 64945 -8.0
8. Aesculin 5281417 -8.5
9. Scopoletin 5280460 -6.4
Standard Drug
10. Artemisinin 68827 -7.5

The binding affinities of phytoconstituents ranged from – 8.9 to -6.4 kcal/mol. From the docked
results, it is evident that the compounds, Nordamnacanthal exhibit the most favourable binding
affinity (−8.9 kcal/mol) and in complex with selected macromolecules (PDB ID:4RAC), as
compared to other docked compounds i.e. Aesculin (−8.5 kcal/mol), Damnacanthal (−8.4
kcal/mol), Rudiadin(−8.2 kcal/mol), Morindone (−8.2 kcal/mol), Lucidin (−8.1 kcal/mol), 2-
benzylzanthopurpurin (−7.8 kcal/mol), Scopoletin (− 6.4 kcal/mol).Visual examination of the
computationally docked optimal binding poses of phytoconstituents on selected
macromolecules (i.e.4RAC) revealed the significant involvement of various types of
interactions, such as hydrogen bonding and hydrophobic interactions, including π–π stacking
and π–alkyl and alkyl interactions, in the stability of the binding of the phytoconstituents to
4RAC
The number of intermolecular hydrogen bonds, the binding energy of ligand 4RAC stable
complexes, and the number of nearest amino acid residues were also determined for selected
compound Artemisinin. All synthesized derivatives formed complexes with target proteins. An
analysis of the interaction between the 4RAC protein complex and the Artemisinin ligand was
also carried out, which showed that the ligand molecule is oriented due to van der Walls
interaction with amino acid residue ILE135, ASP137, THR141, LEU101, GLU133, LEU67,
VAL66, GLX70, GLY69, LYS68, ARG199, AP193 were also found (Fig. 2).
An analysis of the interactions between the 4RAC protein complex and the Nordamnacanthal
ligand was also carried out, which showed that the ligand molecule is oriented due to one Pi–
Pi stacked bond with the PHE186 amino acid fragment, conventional hydrogen bonds
interactions with ASP193, ARG199, LYS68, GLY69, Pi-sigma interaction with IEL135, Pi-
Alkyl interaction with LEU192, Pi-Anion interaction with ASP134. In addition, six van der
Waals interactions with amino acid residues VAL187, LYS165, ASP137, THR141, LEU101,
LEU67 were also found (Fig. 3a).
Analysis of interactions of the 4RAC protein complex and ligand Rubiadin showed that the
ligand molecule is oriented due to one Pi–Pi Stacked bond with the PHE186 amino acid
fragment and to Pi-sigma interactions with amino acid residues ILE135, and forming
conventional hydrogen bonds with ARG199, ASP193, residues and Pi-Alkyl interaction with
LEU192, and Unfavorable Donar interaction with PHE186, and Pi-Anion interaction with
ASP134. In addition, seven van der Waals interactions with amino acid residues VAL187,
LYS165, ASP137, THR141, LEU101, LUE67, GLX69 were also found (Fig. 3b).
An analysis of the interactions between the 4RAC protein complex and the Lucidin ligand was
also carried out, which showed that the ligand molecule is oriented due to one Pi–Pi stacked
bond with the PHE186 amino acid fragment, conventional hydrogen bonds interactions with
the amino acid residues ASP193, LEU67, GLY69 Pi-sigma interaction with IEL135 Pi Alkyl
interaction with LEU192, Unfavorable donar interaction with ARG199, Pi-Anion interaction
with ASP134. In addition, eight van der Waals interactions with amino acid residues VAL187,
LYS165, ASP137, THR141, LEU101, ARG100, VAL66, LYS68 were also found (Fig. 3c).

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Table 2. Binding interactions of ligands with the binding site of Hypoxanthine-guanine

phosphoribosyltransferase (HPRT1)
Main amino acid interaction
Pi-alkyl, Pi-Sigma,
Binding alkyl, Pi-S/Pi-Pi
Sr. No Inhibitors energy H bond stacking/Pi-Pi T van der Waals
(kcal/mol) shaped/halogen/unfavo interaction
urable donor-donor
interactions
VAL187, LYS165,
PHE186, ILE135,
ARG199, ASP137, THR141,
1 Rubiadin -8.2 LEU192, PHE186,
ASP193 LEU101, LEU67,
ASP134
GLY69
VAL187, LYS165,
ASP193, PHE186, ILE135,
ASP137, THR141,
2 Lucidin -8.1 LEU67, LEU192, ARG199,
LEU101, ARG100,
GLY69 ASP134
VAL66, LYS68
VAL187, LYS165,
ASP193, ASP137, THR141,
PHE186, ILE135,
3 Damnacanthal -8.4 ARG199,L LYS102, LEU101,
LEU192, ASP134
YS68 ARG100, LEU67,
GLY69
ASP193,
VAL187, LYS165,
Nordamnacanth ARG199, IEL135, PHE186,
4 -8.9 ASP137, THR141,
al LYS68, LEU192, ASP134
LEU101, LEU67
GLY69
LEU101, LYS102,
2- LEU192, ILE113,
ASP137, LYS165,
5 benzylzanthopur -7.8 ASP193 ILE135, ASP134,
VAL187, ARG199,
purin THR141
GLY69, LYS68
LEU67,
ASP193, VAL187, LYS165,
6 Morindone -8.2 ARG199, PHE186, ASP134 ASP137, GLY69,
LYS68, ARG100, LEU101
LEU192
LYS185, LYS102,
VAL187, LEU101, GLU133,
PHE186, ILE135,
7 Aesculin -8.5 LYS165, LYS68, GLY70,
LEU192, ASP134
ASP193 VAL66, GLY69,
LEU67, ARG199
Standard Drug
ILE135, ASP137,
THR141, LEU101,
No GLU133, LEU67,
8 Artemisinin -7.5 No Interaction
Interaction VAL66, GLY70,
GLY69, LYS68,
ARG199, AAP193

3.2. ADMET study

Pharmacokinetic profile (ADME) and toxicity predictions of the ligands are important attentive
parameters during the transformation of a molecule into a potent drug. In the present study,
these parameters were assessed using SwissADME and pkCSM. The absorption potential and
lipophilicity are characterized by the partition coefficient (Log P) and topological polar surface
area (TPSA), respectively. For better penetration of a drug molecule into a cell membrane, the
TPSA should be less than 140 Å. However, the value of Log P differs based on the drug target.
The ideal Log P value for various drugs are as follows: oral and intestinal absorption, 1.35 −

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1.80; sublingual absorption, > 5; and central nervous system (CNS)32. The aqueous solubility of
ligands ideally ranges from − 6.5 to 0.5 33, while the blood brain barrier (BBB) value ranges
between − 3.0 and 1.2 34. In addition, non-substrate P-glycoprotein causes drug resistance 35.
In our study, all the selected ligands followed the TPSA parameter, P-glycoprotein non-
inhibition, thereby showing good intestinal absorption and an acceptable range of BBB
values. All the compounds showed aqueous solubility values within the range. Further, it was
predicted that the selected ligands do not show AMES toxicity, hepatotoxicity, and skin
sensitivity. In addition, it did not inhibit hERG-I (low risk of cardiac toxicity). Lipinski’s rule
violations, T. pyriformis toxicity, minnow toxicity, maximum tolerated dose, rat acute oral
toxicity, and chronic toxicity are depicted in table 3.

Table 3. ADME and toxicity predicted profile of ligands with superior docking scores

Aqueous Human
Blood-
ADMET MW TPSA HB Hb solubility intestinal
Formula Log P brain
Properties (g/mol) (Ȧ2) doner acceptor (Log absorption
barrier
mol/L) (%)

Rubiadin C15H10O4 254.24 2.18162 74.60 2 4 -3.243 96.938 0.025

Lucidin C15H10O5 270.24 1.3655 94.83 3 5 -2.963 70.752 -0.979
Damnacanthal C15H7O5 267.21 1.0537 94.50 1 5 -3.04 83.174 -0.288
Nordamnacanthal C15H8O5 268.22 1.6857 91.67 2 5 -3.08 92.837 -0.157
2- C21
330.33 3.464 74.60 2 4 -4.133 96.885 0.187
benzylzanthopurpurin H14O4
Morindone C15H10O5 270.24 1.88722 94.83 3 5 -3.093 74.498 -0.707
Ursolic Acid C29H46O3 442.684 6.6994 57.53 2 2 -4.508 96.081 -0.384
Aesculin C15H16O8 324.28 -0.2935 129.59 4 8 -2.482 69.689 -1.298
Scopoletin C10H8O4 192.17 1.5072 59.67 1 4 -2.467 95.015 -0.309
Standard Drug
Artemisinin C15H22O5 282.33 2.3949 53.99 0 5 -3.678 97.543 0.235

Table 3 Continued

Max
Total clearance
ADMET PGP Bioavailability AMES tolerated hERG I hERG II
[Log
Properties substrate score toxicity dose [Log inhibitor inhibitor
mL./(min.kg )]
mg/(kg.d)]
Rubiadin YES 0.129 0.55 YES 0.387 NO NO
Lucidin YES 0.073 0.55 NO 0.761 NO NO
Damnacanthal YES 0.223 0.56 YES 0.077 NO NO
Nordamnacanthal YES -0.024 0.55 NO 0.645 NO NO
2-
YES 0.167 0.55 YES 0.199 NO YES
benzylzanthopurpurin
Morindone YES -0.046 0.55 YES 0.556 NO NO
Ursolic Acid YES 0.13 0.85 NO -0.809 NO NO
Aesculin YES 0.765 0.55 NO 0.0543 NO NO
Scopoletin NO 0.752 0.55 NO 0.292 NO NO
Standard Drug
Artemisinin NO 0.98 0.55 YES 0.065 NO NO

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Table 3 Continued

Oral rat
chronic T. Minnow
Acute oral rat
toxicity Skin Pyriformi toxicity Lipinski’
ADMET toxicity, Hepatotoxicit
(Log sensitisatio s toxicity (Log s rule
Properties LD50(mol/kg y
mg/kg n (Log mmol/L violations
)
bw/day µg/L) )
)
Rubiadin 2.146 1.554 NO NO 0.848 0.364 YES (0)
Lucidin 2.283 2.359 NO NO 0.382 0.457 YES (0)
Damnacanthal 1.998 1.602 NO NO 0.975 0.678 YES (0)
Nordamnacanthal 1.57 2.389 NO NO 0.629 2.071 YES (0)
2-
benzylzanthopurpuri 2.21 1.345 NO NO 0.298 -0.995 YES (0)
n
Morindone 2.124 2.215 NO NO 0.421 1.904 YES (0)
Ursolic Acid 4.018 2.02 YES NO 0.315 -0.229 YES (1)
Aesculin 2.417 3.255 YES NO 0.282 2.865 YES (0)
Scopoletin 2.012 1.424 NO NO 0.453 1.604 YES (0)
Standard Drug
Artemisinin 2.459 1 NO NO 0.322 1.406 YES (0)

3.3 String Pathway

Hypoxanthine-guanine phosphoribosyltransferase (HPRT1): Converts guanine to
guanosine monophosphate, and hypoxanthine to inosine monophosphate. Transfers the 5-
phosphoribosyl group from 5-phosphoribosylpyrophosphate onto the purine. Plays a central
role in the generation of purine nucleotides through the purine salvage pathway.

Fig.2. Network formation of HPRT1 using STRING

3.4 Molinspiration
Lastly, we predicted bioactivity scores to identify the potency of Phyto molecules studied
(Table 4). Our result showed that nordamnacanthal, rubiadin and lucidin might essentially
serve as an interface with Hypoxanthine-guanine phosphoribosyltransferase (HPRT1) as they
acted as enzyme inhibitors. Bioactivity scores of phytoconstituents and standard were
comparable indicating that these phytocompounds might be utilized as an alternative and in
improving treatment to malaria.

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Table 4. Summary of predicted Molinspiration bioactivity score for nordamnacanthal,

rubiadin and lucidin in comparison to Artemisinin
Nuclear
GPCR Ion channel Kinase Protease Enzyme
Bioactivity receptor
ligand modulator inhibitor inhibitor inhibitor
ligand
Nordamnacanthal -0.20 -0.04 0.10 0.15 -0.28 0.22
Rubiadin -0.20 -0.19 -0.03 0.07 -0.26 0.24
Lucidin -0.02 0.04 0.13 0.19 -0.02 0.38
Standard Drug
Artemisinin -0.17 -0.31 -0.65 -0.00 -0.19 0.39

Standard Drug
1. Artemisinin,4RAC

Fig. 3. Docking scores and binding interaction of Artemisinin (PDB ID: 4RAC).
The ligand is shown in line and stick representation along with its 2D diagram and hydrogen
bond interaction.

Drugs to be considered
a. Nordamnacanthal

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b. Rubiadin

c. Lucidin

Fig. 4. Docking scores and binding interaction for antimalarial activity (PDB ID: 4RAC).
The ligand is shown in line and stick representation along with its 2D diagram and hydrogen
bond interaction

Boiled Egg Diagram

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Fig. 5 Combined boiled egg diagram of all phytoconstituents with standard.

Table 5. Molecule names in boiled egg diagram.

MOLECULES NO. DRUG NAME
1 Rubiadin
2 Lucidin
3 Damnacanthal
4 Nordamnacanthal
5 2-benzylzanthopurpurin
6 Morindone
7 Ursolic acid
8 Aesculin
9 Scopoletin
10 Artemisinin

BOILED means Brain Or IntestinaL Estimate D Permeation Predictive model:

The Boiled egg diagram shows two regions white and yellow. The white region is the
physiochemical space of molecules with highest probability of being absorbed by the
gastrointestinal tract, and the yellow region (yolk) is the physiochemical space of molecules
with highest probability to permeate to the brain.
In addition the points are coloured in blue if predicted as activated effluxed by P-gp (PGP+)
An in red if predicted as non-substrate of P-gp (PGP).

CONCLUSION
In this study, we have carried out an in-silico screening of the phytoconstituents
of Hymenodictyon excelsum plant. This study demonstrated the nine compounds from
Hymenodictyon excelsum plant, (Rubiadin, Lucidin, Damnacanthal, Nordamnacanthal, 2-
bezylzanthopurpurin, Morindone, Ursolic Acid, Aesculin, Scopoletin). The selected
phytocompounds showed docking scores ranging from – 8.9 to -6.4 kcal/mol in 4RAC. Among
all, Nordamnacanthal gave the lowest binding energy (− 8.9 kcal/mol) in complex with 4RAC

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whereas the reference compound, Artemisinin, showed a docking score with a binding energy
–7.5kcal/mol. Furthermore, these ligands exhibited good ADMET properties. To summarize,
phytoconstituents present in Hymenodictyon excelsum possess strong effects against
4RAC and could be further evaluated for their anti-malarial effect, as well as for the
development of alternative drugs with fewer side effects for the treatment of malaria.

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70

Spectrophotometric Complexation Study of Cu(II) with 8-

Hydroxyquinoline based Azo dye

Santosh M. Chavan1 Nilesh V. Rathod2, Manoj S. More2, Chandrakant D. Ghugare 1,

Ravi E. Khadse3, Jayshri S. Jadhao2, Akash V. Kubade2 Parikshit S. Thakare2,
Arun B. Patil1
1. Department of Chemistry Phulsing Naik College Pusad MS India-444204
2. Department of Chemistry R.A. Arts Shri. M. K. Commerce and Shri. S.R. Rathi Science College Washim
MS India-444505
3. Late Pundlikrao Gawali Arts and Science College Shirpur Jain MS India -444504
Email: [email protected], [email protected]

Abstract:
In present study Cu(II) and 8-Hydroxyquinoline azo dye (8-HDQ-Cu(II)) were
combined to generate a metal complex, which was then examined using IR, UV-Visible and
Scanning electron microscopic methods. The complex formation spectroscopic peak is visible
at 305 nm. The binding constant and the 1:2 M:L stoichiometry have been ascertained. As a
result, the azo dye of 8-hydroxyquinoline is a commonly used indicator in the complexometric
method to measure Cu(II).

Keywords: 8-Hydroxyquinoline, Cu(II) metal ion, UV-Visible

Introduction:
Potential applications in industry, technology, and life processes have been identified
for coordination complexes containing azo chemicals as ligands[1-4]. Copper complexes have
many different and significant applications. Copper complexes find extensive application as
fungicides, crop defenders, and polymer additives. Metal complex dyes are essential functional
materials that have been incorporated into numerous high-tech applications. The creation of
innovative azo dye structures has drawn attention; many of these structures are beneficial for
commercial applications such as polyester, polyamide, and polyacrylic, as well as blends with
other fibers [5-7]. 8-Hydroxyquinoline (8-HQ), a crucial ligand in this area, is well-known for
its potent coordination skills and special capacity to distinguish between various metals [8]. In
order to use 8-hydroxyquinoline and its derivatives, the phenolate oxygen and quinoline
nitrogen must connect both inside and outside of the cyclic structure. This makes it possible to
use it as a metal chelator because it can bind to metals in bidentate and monomeric forms [9].

Figure 1: 8-Hydroxyquinoline azo dye

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Chemicals and Instruments:

S.D. Fine Chemicals provided all of the chemical substances used in this investigation.
A single beam UV-VIS Bio-Era spectrophotometer with a quartz cell (10 mm) was used to
measure the electronic spectra, and an IR spectrometer made by Brucker was utilized to get the
FT-IR spectra. Additional double-distilled water purification was supplied by Milli-Q. The AR
grade of solvents was used. The 8-HDQ dye stock solution contained 2× 10-4 M, while the
metal solutions had 1× 10-2 M.
Preparation of Ligand and Complex:
The preparation of ligand and its complex was prepared according previously reported
literature [10].

UV-Visible Study of ligand and its Cu(II) complex:

The UV-visible spectroscopic approach was used to characterize the study using a UV-
visible spectrophotometer with a 10 mm quartz cell. This is the spectrum of an ethanol solution
of copper nitrate (5×10-4M) and ligand (5×10-5M) in water after adding some concentrated
nitric acid. It was noted from the graph (Fig.2) that the ligand exhibits its greatest absorbance
at 305 nm, which is the cause of n-π*. However, there is a little shift up to 295 nm that shows
the complex formation during complexation with copper metal ion.

8-HDQ Azo Dye

1.4
8-HDQ Azo Dye-Cu(II)

1.2

1.0
Absorbance

0.8

0.6

0.4

0.2

0.0
240 270 300 330 360 390
Wavelength (nm)

Figure 2: UV-Visible spectra of ligand and its Cu(II) Complex

Metal to ligand stoichiometry:

Cu(II) metal ion solutions and 8-HDQ azo dye were combined in similar mole ratios,
with the dye and metal ion concentrations being constant. The complex's stoichiometry was
ascertained by measuring the absorbance at a wavelength of 305 nm. The relationship between
absorbance and mole fractions is depicted in Figure 3, the resultant absorbance graph. For the
8-HDQ-Cu(II) complexes, the greatest absorbance is observed at a mole ratio of 0.4, indicating
that two ligands are bonded to the metal ion in a 1:2 ratio.

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4.0

3.5

Absorbance @ 305nm
3.0

2.5

2.0

1.5

1.0
0.0 0.2 0.4 0.6 0.8 1.0
Mole Fraction

Figure 3: Jobs plot of Complex fot the stoichiometry

IR Spectra Study:
The FTIR spectra of the developed complexes and ligands are shown in Figure 4. Table
1 displays the spectrum data obtained from FTIR spectroscopy. A broad band of moderate
intensity was seen on the ligands. The O-H bond of 8-HDQ ligands appeared at 3193 cm−1 in
the FTIR spectra, however, it moved to 3106 cm-1 when the complex formed with the Cu(II)
metal ion. In 8-HDQ, a band confirming the presence of the C-O bond was found at 1240 cm-
1
however, in the complexes, these bands migrated to 1230 cm-1. The vibrational frequencies
of the M-O and M-N bonds support the confirmation of the bonding between the ligand and
metal ion. These frequencies are roughly 602 cm-1 for the former and 437 cm-1 for the latter.

Table 1. Infra-red spectral data for 8-HDQ and Co(II) complexes.

Infra red spectral bands ʋ cm-1
Compound OH C-O C-N N=N M-O M-N
8-HDQ 3193 1240 1310 1465 - -
8-HDQ-Cu(II) 3106 1230 1305 1450 610 437

(a)

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(b)
Figure 4: IR spectra of (a) 8-HDQ (b) 8-HDQ-Cu(II) complex
The stability constant of 8-HDQ-Cu(II) complex:
The stability constant, sometimes referred to as a binding constant, is the equilibrium
constant for complex formation in solution. It serves as an indicator of how well the metal and
ligand will bind together to form a complex. The following equation was used to determine the
apparent stability constant (K) of the (1:2) Metal: Ligand complex with the use of spectroscopic
data.
K=1-α/4α3c2 (1)
α =Am-As /Am (2)
or α =AS-Am/As
where, AS is the solution's absorption when ligand and metal ions are present in stoichiometric
proportions, c is the complex solution's concentration in mol/L, and A is the degree of
dissociation. Am=is the quantity of metal absorbed in a solution containing an excess of ligand.
The solution's λmax was used to assess the values of AS and Am. The values of α, K, Am, and
AS were tabulated below Table 2, which demonstrates the high value of K, or 2.3×105. This
outcome demonstrates the strong binding between Cu(II) metal and 8-HDQ azo dye.
Table 2. The stability constant of 8-HDQ-Cu(II) Complex:
complex AS Am α KS
[Bi(L)2] 2.233 2.595 0.01325 2.3×105
Scanning Electron Microscopy Study:
The morphologies of 8-HDQ, and its Cu(II) complexes were investigated in the
scanning electron microscopy (SEM) study. The SEM pictures of the ligands exhibit an
irregular, rock like morphology with varying lateral diameters, as illustrated in Fig. 5(a,b). It's
interesting to note that the complexes' surface shape was significantly changed by metal
coordination to the ligand. In particular, the copper complex's SEM micrographs show a
noticeably uneven structure that resembles broken ice.

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Figure 5: SEM micrograph of (a) 8-Hydroxyquinoline azo dye and (b) 8-HDQ-Cu(II)
complex
Conclusion:
In summary, we have effectively created an 8-hydroxyquinoline compound using Cu(II). UV-
Vis spectroscopy has been used to study Cu(II)binding with 8-HDQ. After research, the
stoichiometry of metal ions and ligands was found to be 1:2. The complex that forms between
copper and the azo dye of 8-hydroxy quinoline has a larger stability constant, indicating that it
is stable. Additional evidence of the binding relationship between Cu(II) and ligand is provided
by SEM studies and IR spectra.

References:
1. B. K. Ghosh and A. Chakravorty, Coord. Chem. Rev. 95, 239 (1989).
2. P. K. Santra, T K Misra, D. Das, C. Sinha, A. M. Z. Salwin, J. D. Woollins, Polyhedron
18, 2869-2878 (1999).
3. S. S. Kandil, Trans. Met. Chem. 23, 461-465 (1998).
4. M. A. Awad, J. Chem. Biotechnol. 53, 227(1992).
5. V.H, Patel, M.P. Patel, R.G. patel, 2002. J. Serb. Chem. Soc 67 (11) : 727-734.
6. M. Mohorcic, J. Friedrich, and A. Pavko, 2004. Acta. Chim. Slov 51(2):619-628.
7. H.R. Maradiya, 2001, Turk. J. Chem 25(6): 441-450.
8. M. Albrecht, M. Fiege, O. Osetska, Coord Chem Rev. 2008;252(8–9):812–824
9. M. Nguyen, L. Vendier, J.L. Stigliani, B. Meunier, A. Robert Eur. J. Inorg. Chem. 2017,
3, 600–608
10. A. El., Nadia, H. F. Wakiel, S.A. Ibrahim, Appl Organometal Chem, 31(10), 1-7, (2017)

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71

A Comprehensive Review on Various Method of Synthesis and Biological

Activities of Schiff Bases with Heterocyclic Moiety

Rahul P. Rahate
Assistant Professor, Department of Chemistry, Arts, Science and Commerce, College, Chikhaldara,
Maharashtra, India, [email protected]

Abstract:

Schiff bases are versatile chemical compounds that are becoming more important because of
their numerous applications. Schiff bases, which include imines or azomethine functional
groups, are formed by the condensation of primary amines with carbonyl compounds or can
exist naturally in plants. They are widely used in industry and exhibit a variety of biological
activities, such as antibacterial, antifungal, antiviral, and anticancer properties. Researchers are
presently focusing on a wide range of biological studies of Schiff bases, which could lead to
the discovery of new lead compounds. This review covers discoveries and various methods for
synthesising specially Schiff bases with heterocyclic moiety, as well as their biological
functions.

Keywords: Schiff bases; Biological activities; Applications; Antifungal; Antiviral;

Anticancer

Introduction
Hugo Schiff (1834–1915) (Figure 1) was a German scientist. He
discovered some bases and named them Schiff bases [1]. Schiff bases
are generated by reacting a primary amine with carbonyl compounds
(aldehydes or ketones) under specific conditions. The common
structure is R1R2C=NR (R ≠ H), with the main function being the imine
or azomethine (-C=N-) group (Figure 2) [2].
Figure 1.
A portrait of
Hugo Schiff
Schiff bases, particularly those coupled to a heterocyclic moiety,
displayed a wide range of pharmacological and biological actions, including antibacterial,
cytotoxic, antifungal, antimalarial, anticonvulsant, antioxidant, and anti-inflammatory
properties [3–10]. Schiff base used in different field given below in Fig.3.
R1
R1 CHO
N R
Aldehyde
R NH2 + -H20
Amine H
OR OR
R2
R1
O
N R
R1
Ketones R2
Schiff Bases
Figure 2 R1,R2 and/or R2= Alkyl or Aryl group
Fig. 3
Various Synthesis Methods for Schiff Bases: Schiff bases have been employed in a variety
of industries. As a result, many approaches and innovative procedures for the preparation of
Schiff bases have been documented, including the following:

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i) Conventional or common heating method:

The Schiff base containing benzothiazole moiety 3 was made by reacting 2-amino-6-
methylbenzothiazole 1 with 5-bromo-2-hydroxybenzaldehyde 2 in ethanol as a solvent and
refluxing the reaction mixture for two hours. [11] (Scheme 1).

N NH2 HO N

+ N N
NH Absolute EtOH
Reflux 12-24 H N
1 2
R N HO
R R= CN/CH3 O H 3

Scheme 1
ii) Using water as a solvent:
Mono-Schiff bases 5 and 6 were prepared by the stirring of 1,2-diaminobenzene 4 with
thiophene-2-carbaldehyde and 1H- H2N N
pyrrole-2-carbaldehyde, O
S
respectively, in H2O as a S

solvent.[12] H 2 O/R.T.
H2N NH2
5

H2 N N
O
N NH
4 H
H2O/R.T
6

SScheme 2
iii) Metal catalyzed:
Cerium (III) chloride catalyzed: Schiff base 9 was produced by the reaction of aromatic
amines 7 with aldehydes 8 in the presence of CeCl3.7H2O as a catalyst under solvent-free
conditions. [13] (Scheme 3).
O
NH2
CeCl3. 7H2O N
+ R1
Solvent Free
R 8 R1
7
9
R
R= H,2-OH,4-OCH3 R1=H,4-NO2,3-NO2,4-Cl

Scheme 3
iv) Acidic and phase transfer catalyst (PTC) conditions:
The reaction of 2-amino-5-mercapto-1,3,4-thiadiazole 10 with aromatic aldehydes 11 in
ethanol in the presence of H2SO4 (acidic conditions) produced 1,3,4-thiadiazole Schiff bases
12. Benzyl triethylammonium chloride (BTEAC) was also used as a catalyst during the
process solvent-free [14] (Scheme 04)
O

N N N
EtOH/H2SO4/Reflux
N
+ N
NH2 Or BTEAC/Solvent Free
HS S S
10 11 HS
R 12
R
R= 2-OCH3,2-Cl,3-Br,4-NMe,4-F,4-OCH2Ph

v) Ultrasonic and microwave conditions

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Chiral-Schiff bases 15 were synthesized by ultrasonically reacting 2-naphthaldehyde 13 with

chiral α- amino acids 14 in the presence of K2CO3.[15]
HOOC
H
H2N COOH
O K3CO3,H2O/EtOH
+ N
Ultrasonic conditions, RT R
R H
13 14
Amino Acids = L-Valine,L-Phenyl alanine, L-Tyrosine
L-Aspartic Acid, L-Methionine 15

Scheme 5
vi) Grinding chemistry technique:
The synthesis of bioactive chemicals was carried out using grindstone technology.
Furan-2-carbaldehyde 16 and DL-alanine 17 were reacted in water as a green solvent to
produce Schiff base (E)-2-(furan-2-ylmethyleneamino) propanoic acid 18 [16].
CH3
H3C O
Grinding
+
H2O N O
O O H2N OH O
16 17 18 HO
Scheme 6
Biological activities of Schiff bases :
Antimicrobial activities:
Triazole-Schiff bases 19 showed strong antibacterial activity against Escherichia coli,
Salmonella typhi, and Bacillus subtilis. In addition, they had significant antifungal activity
against Candida albicans, Aspergillus flavus, Fusarium solani, and Candida glabrata. [17]
(Figure 4).
Isatin-Schiff base 20 showed significant antibacterial action against pseudomonas
aeruginosa (MIC = 6.25 mg/mL) [18] (Figure 5).
Acetylenic indole-Schiff base 21 shown antibacterial activity against Staphylococcus aureus,
with a MIC of 7.81 µM. Indole-Schiff base 22 showed antifungal efficacy against Candida
krusei, with a MIC of 15.62 µM [19] (Figure 6).

HO
N
H
H3CS
N
N
N N
N H
N O
N 19 R H
H O 20
Fig. 5
R=H,Cl,Br,NO2
Fig. 4 Triazole-Schiff bases

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OCH3

N
N

N
N
22
21

Figure 6. Acetylenic indole-Schiff bases

Piperazine-sulphonamide-linked to Schiff base 23 showed potent antibacterial activity against

Bacillus subtilis with MIC= 26.1 µg/mL [20] (Figure 7).
N Cl
O
N
S
N
O

23
OEt
Figure 7 Piperazine-sulphonamide linked to Schiff base 6
Anticancer activities:
1,3,5-Triazine-isatin Schiff base 24 showed anticancer activities against lung (HOP-
92), leukemia (CCRF-CEM), and leukemia (SR) cancer cell lines [21] (Figure 8).
Cl

N N

N
Cl N N
H

NH
O
Fig.8 1,3,5-Triazine-isatin Schiff base 24
Anti-inflammatory activities:
Schiff base based on quinazolin-4-one in combination with 1, 3, 4-oxadiazole moiety
25 showed anti-inflammatory effects [22] (Figure 9).
HO

N
N

N O

O N
N
Fig. 9 Schiff base based on quinazolin-4-one with 1,3,4-oxadiazole 25
Analgesic activities: Isatin-Schiff base 26 exhibited good analgesic activity [23] (Figure 10).

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N
OH

N O
H
OCH3
Figure 10 . Isatin-Schiff base 26

Anthelmintic activities:
Antipyrine-coumarin linked to Schiff bases 27 showed excellent anthelmintic activities
[24] (Figure 11).
HO
O

N CH3

N
O
N
CH3
H3C
O

Figure 11 Antipyrine-coumarin linked to Schiff bases 27

Antioxidant activities:
Halogenated Schiff bases 25 showed very low to
moderate antioxidant activities [24] (Figure 12). OH

Conclusion: N
Schiff bases were characterized by the presence of the F R
imine or azomethine (–C=N–) group. This review focused
on some synthesis and biological activities of Schiff bases. H3C
From this review, it can be concluded that Schiff bases R= C3H7,C5H11,C6H13,C7H15
especially Schiff bases-heterocyclic moiety conjugates Figure 12. Halogenated Schiff bases 28
display a wide range of pharmacological activities. For that,
Schiff bases attracted increasing attention to the scientists for the synthesis of new derivatives
for applications in medicinal and in industrial field.

References:
1. Qin, W., Long, S., Panunzio, M., & Biondi, S. (2013). Schiff bases: A short survey on an evergreen
chemistry tool. Molecules, 18(10), 12264-12289.
2. Ghosh, P., Dey, S. K., Ara, M. H., Karim, K., & Islam, A. B. M. (2019). A review on synthesis and versatile
applications of some selected Schiff bases with their transition metal complexes. Egyptian Journal of
Chemistry, 62(Special Issue (Part 2) Innovation in Chemistry), 523-547.
3. Antony, R., Arun, T., & Manickam, S. T. D. (2019). A review on applications of chitosan-based Schiff
bases. International journal of biological macromolecules, 129, 615-633.
4. Ledeţi, I., Alexa, A., Bercean, V., Vlase, G., Vlase, T., Şuta, L. M., & Fuliaş, A. (2015). Synthesis and
degradation of Schiff bases containing heterocyclic pharmacophore. International Journal of Molecular
Sciences, 16(1), 1711-1727.
5. Pund, A. A., Saboo, S. S., Sonawane, G. M., Dukale, A. C., & Magare, B. K. (2020). Synthesis of 2, 5-
disubstituted-1, 3, 4-thiadiazole derivatives from (2S)-3-(benzyloxy)-2-[(tert-butoxycarbonyl) amino]
propanoic acid and evaluation of anti-microbial activity. Synthetic Communications, 50(24), 3854-3864.
6. Magalhães, T. F. F., da Silva, C. M., Dos Santos, L. B. F., Santos, D. A., Silva, L. M., Fuchs, B. B., ... & de
Fátima, Â. (2020). Cinnamyl Schiff bases: Synthesis, cytotoxic effects and antifungal activity of clinical
interest. Letters in Applied Microbiology, 71(5), 490-497.
7. Sztanke, K., Maziarka, A., Osinka, A., & Sztanke, M. (2013). An insight into synthetic Schiff bases

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revealing antiproliferative activities in vitro. Bioorganic & Medicinal Chemistry, 21(13), 3648-3666.
8. Hassan, A. S., Askar, A. A., Naglah, A. M., Almehizia, A. A., & Ragab, A. (2020). Discovery of new Schiff
bases tethered pyrazole moiety: Design, synthesis, biological evaluation, and molecular docking study as
dual targeting DHFR/DNA gyrase inhibitors with immunomodulatory activity. Molecules, 25(11), 2593.
9. Morsy, N. M., Hassan, A. S., Hafez, T. S., Mahran, M. R., Sadawe, I. A., & Gbaj, A. M. (2021). Synthesis,
antitumor activity, enzyme assay, DNA binding and molecular docking of Bis-Schiff bases of
pyrazoles. Journal of the Iranian Chemical Society, 18, 47-59.
10. Murtaza, G., Mumtaz, A., Khan, F. A., Ahmad, S., Azhar, S., Najam-Ul-Haq, M., ... & Hussain, I. (2014).
Recent pharmacological advancements in schiff bases: A review. Acta Pol. Pharm, 71(4), 531-535.
11. Huda, A. S., Asia, A. S., & Zainab, W. S. (2020). INTERNATIONAL JOURNAL OF RESEARCH IN
PHARMACEUTICAL SCIENCES.
12. Rao, V. K., Reddy, S. S., Krishna, B. S., Naidu, K. R. M., Raju, C. N., & Ghosh, S. K. (2010). Synthesis of
Schiff's bases in aqueous medium: a green alternative approach with effective mass yield and high reaction
rates. Green Chemistry Letters and Reviews, 3(3), 217-223.
13. Ravishankar, L., Patwe, S. A., Gosarani, N., & Roy, A. (2010). Cerium (III)-catalyzed synthesis of schiff
bases: a green approach. Synthetic Communications®, 40(21), 3177-3180.
14. Mobinikhaledi, A., Jabbarpour, M., & Hamta, A. (2011). Synthesis of some novel and biologically active
Schiff bases bearing a 1, 3, 4-thiadiazole moiety under acidic and PTC conditions. Journal of the Chilean
Chemical Society, 56(3), 812-814.
15. Şendil, K., Tekin, T., Göksu, H., Oğuz, M., Anıl, B., & Gültekin, M. S. (2016). A Novel Method for the
Synthesis of Newfangled Asymmetric Schiff Bases from α‐amino Acids under Ultrasonic Conditions and
in Aqueous Medium. Journal of the Chinese Chemical Society, 63(9), 808-817.
16. Sachdeva, H., Saroj, R., Khaturia, S., & Dwivedi, D. (2012). Operationally simple green synthesis of some
Schiff bases using grinding chemistry technique and evaluation of antimicrobial activities. Green
Processing and Synthesis, 1(5), 469-477.
17. Chohan, Z. H., & Hanif, M. (2013). Antibacterial and antifungal metal based triazole Schiff bases. Journal
of Enzyme Inhibition and Medicinal Chemistry, 28(5), 944-953.
18. Tehrani, K. H. M. E., Hashemi, M., Hassan, M., Kobarfard, F., & Mohebbi, S. (2016). Synthesis and
antibacterial activity of Schiff bases of 5-substituted isatins. Chinese Chemical Letters, 27(2), 221-225.
19. Singh, G., Kalra, P., Arora, A., Singh, A., Sharma, G., Sanchita, ... & Verma, V. (2018). Acetylenic Indole‐
Encapsulated Schiff Bases: Synthesis, In Silico Studies as Potent Antimicrobial Agents, Cytotoxic
Evaluation and Synergistic Effects. ChemistrySelect, 3(8), 2366-2375.
20. Patil, R. H., Kalam Khan, F. A., Jadhav, K., Damale, M., Akber Ansari, S., Alkahtani, H. M., ... &
Sangshetti, J. N. (2018). Fungal biofilm inhibition by piperazine‐sulphonamide linked Schiff bases: Design,
synthesis, and biological evaluation. Archiv der Pharmazie, 351(3-4), 1700354.
21. Polovkovych, S. V., Karkhut, A. I., Marintsova, N. G., Lesyk, R. B., Zimenkovsky, B. S., & Novikov, V.
P. (2013). Synthesis of New Schiff Bases and Polycyclic Fused Thiopyranothiazoles Containing 4, 6‐
Dichloro‐1, 3, 5‐Triazine Moiety. Journal of Heterocyclic Chemistry, 50(6), 1419-1424.
22. Dewangan, D., Nakhate, K. T., Verma, V. S., Nagori, K., & Tripathi, D. K. (2017). Synthesis,
Characterization, and Screening for Analgesic and Anti‐Inflammatory Activities of Schiff Bases of 1, 3, 4‐
Oxadiazoles Linked With Quinazolin‐4‐One. Journal of Heterocyclic Chemistry, 54(6), 3187-3194.
23. Chinnasamy, R. P., Sundararajan, R., & Govindaraj, S. (2010). Synthesis, characterization, and analgesic
activity of novel schiff base of isatin derivatives. Journal of advanced pharmaceutical technology &
research, 1(3), 342.
24. Manjunath, M., Kulkarni, A. D., Bagihalli, G. B., Malladi, S., & Patil, S. A. (2017). Bio-important
antipyrine derived Schiff bases and their transition metal complexes: Synthesis, spectroscopic
characterization, antimicrobial, anthelmintic and DNA cleavage investigation. Journal of Molecular
Structure, 1127, 314-321.
25. Singh, B. B., Shakil, N. A., Kumar, J., Rana, V. S., & Mishra, A. (2016). Microwave synthesis,
characterization, and bio-efficacy of novel halogenated Schiff bases. Journal of Environmental Science and
Health, Part B, 51(8), 558-570.

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72

Revolutionizing Applications: A Comprehensive Review of Schiff Base

Nanoparticles at the Nanoscopic Scale

S. R. Khandekar
Department of Chemistry, Indira Mahavidyalaya, Kalamb, Dist. Yavatmal, (MH) India.
Email – [email protected]

Abstract
Nanotechnology has emerged as a transformative field with the potential to revolutionize
various scientific and technological domains. This chapter delves into the nanoscopic marvels
of Schiff base nanoparticles and their applications in advanced technologies. Schiff bases,
known for their versatile coordination chemistry, have been harnessed to engineer
nanoparticles with unique properties and functionalities. The exploration of these nanoscale
wonders encompasses synthesis methodologies, characterization techniques, and their diverse
applications in fields such as catalysis, drug delivery, sensing, and imaging. The chapter
discusses the tailored design of Schiff base nanoparticles to achieve enhanced performance,
stability, and selectivity in various applications. Moreover, it highlights the potential impact of
these nanomaterials on advancing technologies and offers insights into future directions for
research and development in this rapidly evolving field. The intricate interplay between
molecular design, synthesis strategies, and applications underscores the significance of Schiff
base nanoparticles as promising candidates for addressing contemporary challenges in
nanotechnology.
Keywords: Schiff base nanoparticles, catalysis, drug delivery, nanotechnology.
1. Introduction
The imine (-C=N-) functional group that results from the interaction of the amine amino group
with the aldehyde or ketone carbonyl group defines the Schiff bases class of chemical
compounds [1].The imine functionality of Schiff bases confers significant chemical and
biological features, enabling these molecules to coordinate and complexate with metal ions in
a variety of chemistry-related domains [2].Schiff base compounds can function as ligands to
produce transition metal Schiff base complexes because of the imine group [3] . Additionally,
the imine group (>C = N −) of SB ligands stabilizes the nanoparticles by forming bonds with
them [4]. The term nanotechnology was first used in 1974 by Japanese scientist Norio
Taniguchi and is considered a marvel of science, engineering, and technology [5-7].The word
"nano" comes from the Greek "nanos," which means "small" (10–9 m). Nanoparticles (NPs) are
essentially very small particulates that range in size from 1 to 100 nm [8].A higher level of the
NPs is ensured by the surface modification, increasing their potential in the disciplines of
analysis, electro analytical, catalysis, and biology. SB modified NPs are applied as biosensor,
adsorbent, electro catalysis, catalytic dye degradation, and oxidation of different of various
organic compounds [9-14].This chapter delves into the intricate realm of Schiff base
nanoparticles, investigating their unique properties and potential applications in various
advanced fields.
2. Schiff base complex as a heterogeneous catalyst:
A Cu(II)-Schiff base complex supported on functionalized Fe3O4 magnetic nanoparticles
(MNPs@Salen-Cu(II)) was firstly prepared by the refluxing it in the toluene solvent for two
days. Further this was stirred in ethanol at room temperature with the addition of benzaldehyde
and nitromethane. These nanoparticles were employed as effective catalysts in the manufacture
of derivatives of nitroaldol through the reaction of nitromethane with aldehydes (Scheme 1).
The reaction benefits include selectivity to products, recyclability of the catalysts, simplicity

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in workup, facile separation, cleanliness, and eco-friendliness. Using VSM, SEM, FT-IR, 1H
NMR, XRD, TGA, and EDX, the produced catalysts were characterized. Using an external
magnet, the nanocatalyst may be easily extracted from the reaction mixture and reused 14 times
without experiencing a substantial reduction in catalytic activity. Research on the catalyst's
leaching revealed that the heterogeneous catalyst's active components do not leach [15].

Scheme 1. Synthesis of nitroaldols by the MNPs@Salen-Cu(II)

3. Exploring the Multifaceted Applications of Nickel Nanoparticles Stabilized by Schiff
Base Ligands:
Paulraj Adwin Jose et al. synthesized nickel nanoparticles stabilized by Schiff base ligands
derived from pyrimidine derivatives and are verified by using spectroscopic and microscopic,
SEM and TEM techniques (Scheme 2). The synthesized compounds exhibited strong DNA
binding capability and binding through groove mode. The produced compounds enhanced
DNA denaturation and the BSA binding investigations demonstrated the compound's strong
BSA interaction. The DMPMM and DMPMM-NiNPs were the subject of antimicrobial
investigations using the Agar-Agar well diffusion method. The results of anticancer
experiments verified that the produced DMPMM-NiNPs had a specific characteristic of
interacting with cancer cells as opposed to normal cells. However the activity rose to a level
above that of regular cisplatin when it was capped with metal nanoparticles. The results of tests
on antioxidant scavenging demonstrated the strong antioxidant activity of DMPMM-NiNPs
[16].

Scheme 2. Synthesis of Schiff base ligand capped nickel nanoparticles (DMPMM-NiNPs).

4. Schiff-Bases with Anisaldehyde or Salicylaldehyde in the Presence of Silver
Nanoparticles:
Carboxymethyl chitosan-Schiff-bases (CMCs-SB) of anisaldehyde or salicylaldehyde in the
absence/presence of silver nanoparticles; with different weight ratios of AgNO3 (2 and 5% by
weight) were prepared by Reham A. Abdel-Monem et al. (Scheme 3) and evaluated as
materials with antibacterial efficiencies. The material is characterized by FTIR, SEM, TEM,
TGA.The two carboxymethyl chitosan Schiff-bases nanocomposites were found to exhibit
inhibitory power against the growth of both Gram +ve and Gram -ve types of bacteria. The
antibacterial efficiency of salicylaldehyde/CMCs-SB/AgNPs is higher than that of
anisaldehyde/CMCs-SB/AgNPs. The human hepatocellular carcinoma cell line (HEPG2) and

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the breast adenocarcinoma (MCF7) were used to test the cytotoxicity of the produced CMC
Schiff bases in the presence and absence of AgNPs (2 and 5%). The findings demonstrated that
adding AgNPs at weight ratios of 2 and 5 % enhanced the anticancer activity of the produced
Schiff bases [17].

Scheme 3. Schematic diagram for the reaction of CMCs with anisaldehyde or salicyladehyde.
Leila Zare Fekri et al. synthesized a copper/Schiff base complex immobilized on amine-
functionalized silica MMNPs (Scheme 4). Product is characterized by SEM, TEM, TGA and
VSM analysis. The synthesized product is an effective catalyst in a multicomponent reaction
involving amine, thioglycolic acid, and aldehyde while stirring. This nanoparticle is a novel
mix of organic and inorganic materials. The key benefits of this catalytic process include waste
reduction, good selectivity, easy separation and recycling of the magnetic catalyst, outstanding
product yields, and low operating complexity. Additionally, this new route is affordable and
safe for the environment [18].

Scheme.4 Synthesis of Fe3O4@SiO2@KIT-6-NH2@Schiff base complex

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5. Exploring Anticancer Potential through DNA Binding by Zn(II) Complex with Schiff
Base Ligand:
The study involves the synthesis of biologically potent Pt (II) metal ion complexes,
characterized by a proposed square planar environment through various physicochemical
investigations. These complexes exhibit higher antimicrobial activity compared to the original
ligands and demonstrate a strong ability to cleave DNA via the oxidative nuclease pathway,
confirmed by band tailing in the DNA spectrum. Additionally, the compounds show significant
in vitro anticancer activity against human breast cancer (MCF-7) cell lines, with results
compared to the standard drug Cisplatin. The Pt (II) complex induces DNA damage, as
evidenced by increased cell cycle depletion of S and G2/M at various concentrations, leading
to G1 phase arrest. A notable rise in sub-G1 cells indicates that the complex causes sufficiently
high DNA damage, resulting in a large number of cells undergoing apoptosis [19].
Charles, A. et al. synthesized the ZnO film's polycrystalline nature, with its hexagonal
wurtzite structure and (002) preferred orientation (Scheme 5). The compound is characterized
by Atomic force microscopy (AFM), optical research, and X-ray diffraction (XRD). It was
calculated that the grain size was in the nm range. The direct bandgap value of 3.2 eV in the
produced films is in close agreement with the stated value of 3.37 eV found in literature. The
characterisation studies verify that the deposited ZnO thin films are suitable for use in solar
cells and optoelectronic devices [20].

Scheme 5: Synthesis of N-((1H-pyrrol-2-yl)methylene)-4-methoxyaniline Schiff

base(PMMA) and its copper complex [Cu(PMMA)2]Cl2.2H2O.

A novel compound of Zn(II) was synthesized and characterized using a metal with a
molar ratio of 1:2 and a Schiff base ligand by Masumeh Galini et al. They used FT-IR, 1H-
NMR, UV-Vis X-ray data to determine the structure of the synthesized compound.
Furthermore, ZnO nanoparticles of [ZnL(2-aminopyridine)Br] using the combustion synthesis
method was created Using FESEM, TEM, FT-IR, and XRD, the shape and crystalline structure
of the produced zinc oxide nanoparticles were examined. Additionally, it was shown through
the use of molecular docking techniques that the Zn(II) complex binds to the DNA helix's
minor groove. [ZnL(2-aminopyridine)Br] will function as a tyrosine kinase inhibitor to treat
cancer by blocking the receptor for the epidermal growth factor [21].

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Scheme 6. Synthetic procedure for the preparation of Schiff base ligand and complex (2:1)
and protocol of ZnO nanoparticles.
6. Conclusions
In conclusion, the exploration of Schiff Base Nanoparticles in this review highlights their
remarkable potential for advanced applications at the nanoscopic level. The synthesis and
characterization of these nanoparticles, with a focus on their unique properties, have paved the
way for diverse applications in various fields. From enhanced antimicrobial activity and DNA
cleavage capabilities to potent in vitro anticancer effects, Schiff Base Nanoparticles emerge as
versatile candidates for innovative biomedical and materials science applications. As we delve
into the intricacies of their physicochemical properties and biological activities, it becomes
evident that Schiff Base Nanoparticles offer a promising avenue for addressing complex
challenges in medicine, materials science, and beyond. The observed cell cycle depletion, G1
phase arrest, and induction of apoptosis underscore their potential as impactful agents in cancer
therapeutics. The integration of Schiff Base Nanoparticles into the realm of nanotechnology
signifies a frontier in the ongoing pursuit of advanced materials with tailored functionalities.
Further research and development in this area are essential to unlock the full spectrum of
possibilities and overcome potential challenges. As we embark on this journey, the nanoscopic
marvels of Schiff Base Nanoparticles hold immense promise for shaping the future of advanced
applications, offering unprecedented solutions for a myriad of scientific and technological
endeavors.
References
[1] Berhanu, A.L.; Gaurav; Mohiuddin, I.; Malik, A.K.; Aulakh, J.S.; Kumar, V.; Kim, K.-H. A review
of the applications of Schiff bases as optical chemical sensors. TrAC Trends Anal. Chem., 116, (2019)
74–91.
[2] Soufeena, P.P.; Nibila, T.A.; Aravindakshan, K. Coumarin based yellow emissive AIEE active
probe: A colorimetric sensor for Cu2+ and fluorescent sensor for picric acid. Spectrochim. Acta Part A
Mol. Biomol. Spectrosc., 223, (2019) 117201.
[3] Aytac, S.; Gundogdu, O.; Bingol, Z.; Gulcin, ˙I. Synthesis of Schiff Bases Containing Phenol Rings
and Investigation of Their Antioxidant Capacity, Anticholinesterase, Butyrylcholinesterase, and
Carbonic Anhydrase Inhibition Properties. Pharmaceutics, 15, (2023) 779.
[4] P.A. Jose, M. Sankarganesh, J.D. Raja, G.S. Senthilkumar, R.N. Asha, S.J. Raja, C. D. Sheela, Bio-
inspired nickel nanoparticles of pyrimidineSchiff base: In vitro anticancer, BSA and DNA interactions,
molecular docking and antioxidant studies, J. Biomol. Structure, Dynamics (Pembroke, Ont.) (2021) 1–
15
[5] Y. Wang, A. Chinnathambi, O. Nasif, S.A. Alharbi, Green synthesis and chemical characterization
of a novel anti-human pancreatic cancer supplement by silver nanoparticles containing Zingiber
officinale leaf aqueous extract, Arabian Journal of Chemistry 14 (2021), 103081.

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[6] E.E. Elemike, E.O. Dare, I.D. Samuel, J.C. Onwuka, 2-Imino-(3,4-dimethoxybenzyl) ethanesulfonic
acid Schiff base anchored silver nanocomplex mediated by sugarcane juice and their antibacterial
activities, J. App. Res. Tech. 14 (2016) 38–46.
[7] K. Kalimuthu, B.S. Cha, S. Kim, K.S. Park, Eco-friendly synthesis and biomedical applications of
gold nanoparticles: A review, Microchemical Journal 152 (2020) 104296.
[8] M. Sajida, J.P. Wasylka, Nanoparticles: Synthesis, characteristics, and applications in analytical and
other sciences, Microchemical Journal 154 (2020), 104623.
[9] S. Kannaiyan, Easwaramoorthy, K. Kannan, V. Andal, Andal, Synthesis, characterisation, and
antimicrobial efficacy of acid fuchsin Schiff base-modified silver nanoparticles, Nanotech. Russia 15
(11-12) (2020) 828–836.
[10] S. Moradinasab, M. Behzad, Removal of heavy metals from aqueous solution using Fe3O4
nanoparticles coated with Schiff base ligand, Desalination, Water Treat. 57 (9) (2016) 4028–4036.
[11] R. Mehdaoui, L. Chaabane, E. Beyou, M. Hassen, V. Baouab, Sono-heterogeneous Fenton system
for degradation of AB74 dye over a new tetraaza macrocyclic Schiff base cellulose ligand-loaded
Fe3O4 nanoparticles, J. Iran. Chemi. Soci. 16 (2019) 645–659.
[12] J.M. Abad, I. Bravo, F. Pariente, E. Lorenzo, Multi-tasking Schiff base ligand: a new concept of
AuNPs synthesis, Analytical and Bioanalytical Chemistry 408 (9) (2016) 2329–2338.
[13] S. Rayati, E. Khodaei, M. Jafarian, A. Wojtczak, Mn-Schiff base complex supported on magnetic
nanoparticles: Synthesis, crystal structure, electrochemical properties and catalytic activities for
oxidation of olefins and sulfides, Polyhedron (2017).
[14] S.A. Hamrahian, J. Rakhtshah, S.M.M. Davijani, S. Salehzadeh, Copper Schiff base complex
immobilized on silica-coated Fe3O4 nanoparticles: a recoverable and efficient catalyst for synthesis of
polysubstituted pyrroles, Appli. Organometal. Chem. (2018) 1–12.
[15] Niloufar Parandeh-Khoozani and Mohsen Moradian, Synthesis of nitroaldols through the Henry
reaction using a copper(II)–Schiff base complex anchored on magnetite nanoparticles as a
heterogeneous nanocatalyst, J Coord Chem .,74 (12) (2021) 2035–2054.
[16] P. A. Jose, M. Sankarganesh, J. D. Raja, G. S. Senthilkumar, R. N. Asha, S. J. Raja & C. D. Sheela,
Bio-inspired nickel nanoparticles of pyrimidine-Schiff base: In vitro anticancer, BSA and DNA
interactions, molecular docking and antioxidant studies, Journal Of Biomolecular Structure And
Dynamics (Taylor and Francis), 40 (21) (2022) 10715-10729.
[17] Reham A. Abdel-Monem, Ahmed M. Khalil, Osama M. Darwesh, Ahmed I. Hashim & Samira T.
Rabie, Antibacterial properties of carboxymethyl chitosan Schiff-base nanocomposites loaded with
silver nanoparticles, J. Macromol. Sci. A., (Taylor and Francis), 57 (2) (2020) 145-155.
[18] Leila Zare Fekri, Shohreh Zeinali, Copper/Schiff-base complex immobilized on amine
functionalized silica mesoporous magnetic nanoparticles under solvent-free condition: A facile and new
avenue for the synthesis of thiazolidin-4-ones, Appl Organometal Chem. (2020) e5629.
[19] Bhavana Gupta, Anita Kumari, Savita Belwal, R. V Singh & Nighat Fahmi, Synthesis,
characterization of platinum(II) complexes of Schiff base ligands and evaluation of cytotoxic activity
of platinum nanoparticles, Inorganic and Nano-Metal Chemistry, 50:10, (2020) 914-925.
[20] Charles, A., Sivaraj, K., Krishnaraj, S., Synthesis of Copper(II) Schiff Base Complex and Its Mixed
Thin Layer with ZnO Nanoparticles, Iran. J. Chem. Chem. Eng., 40 (3) (2021) 758-764.
[21] Masumeh Galini, Mehdi Salehi, Maciej Kubicki, Mehdi Bayat, Rahime Eshaghi Malekshah,
Synthesis, structural characterization, DFT and molecular simulation study of new zinc-Schiff base
complex and its application as a precursor for preparation of ZnO nanoparticle, Journal of Molecular
Structure, 1207 (2020) 127715.

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73

Screening Of Antioxidant Property Present In Psidium Guajava Linn

V D Mane & P P Mhasal.

Department of Chemistry,
Shankarlal Khandelwal Arts, Science and Commerce College, Akola.( 444 002 ).
[email protected]

Abstract:
Psidium guajava(Linn) is a member of Myrtaceae family, is a common tropical plant with a
long history of traditional usage. Psidium guajava have been reported to possess variety of
biology activity. Antioxidants both are natural and man-made substance that protects our cell
from free radical. In present study we find out the antioxidant and phytochemical property of
and water extract of Psidium guajava leaves by using DPPH as a free radical scavenger and
colorimeter. The activity was evaluated by the decrease in absorbance as the result of DPPH
colour change from purple to yellow. The higher the sample concentration used, the stronger
was the free radical-scavenging effect. From this study we found that the Psidium guajava
(linn) has good Antioxidant property. This study revealed that guava leaf extracts comprise
effective potential source of natural antioxidants.

Key word: Psidium guajava leaves antioxidant property, DPPH.

Introduction
The guava is a small tree with tortuous branches and a smooth trunk. The fruit is a berry
with a fleshy yellow or pink pulp and many seeds near the center. Many different varieties have
been developed that vary in colour, size and in shape from round to ovoid to pear – shaped.
The guava (Psidium guajava L.) tree , belonging to the Myrtaceae family, is a very unique and
traditional plant which is grown due to its diverse medicinal and nutritive properties. Guava
has been grown and utilized as an important fruit in tropical areas like India, Indonesia,
Pakistan, Bangladesh, and South America. Different parts of the guava tree, i.e., roots, leaves,
bark, stem, and fruits, have been employed for treating stomachache, diabetes, diarrhea, and
other health ailments in many countries. Guava leaves (Psidii guajavae folium; GL) are dark
green, elliptical, oval, and characterized by their obtuse-type apex. Guava leaves, along with
the pulp and seeds, are used to treat certain respiratory and gastrointestinal disorders, and to
increase platelets in patients suffering from dengue fever. GLs are also widely used for their
antispasmodic, cough sedative, anti-inflammatory, antidiarrheic, antihypertension, antiobesity,
and antidiabetic properties. Studies on animal models have also established the role of GL
isolates as potent antitumor, anticancer, and cytotoxic agents.

Material and method:

The guava leaves were collected and shade dried at room temperature and coarsely powdered
using an electric grinder. The coarse powdered materials of leaves were kept in the airtight
polythene bag and stored in dry place. The powdered guava leaves were then extracted in
distilled water.
Solvent extraction
Preparation of Aqueous extract of guava leaves.

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Aqueous extracts of powdered guava leaves was prepared by boiling method. About 10 gram
of powder of guava leaves was taken in 50 gram of distilled water and was warmed for about
10 min, and solution was then allowed to cool and filtered through a filter paper. The filtrate
then was used for studying the antioxidant activity.

Phytochemical investigation of psidium guajava,linn.

Sr. no Constituent Aqueous Extract

1 Carbohydrate +

2 Tannins +
3 Saponins -
4 Flavonoids +
5 Alkaloids -
6 Phenols +
7 Glycosides +
8 Proteins -
9 Sterols +
10 Anthocyanin -

Study of qualitative antioxidant activity of psidium guajava linn.

Freshly prepared 0.02% of DPPH solution in ethanol was prepared. Single of Aqueous extract
of guava leaves was taken on TLC plates, after drying the spots the TLC plates were dipped in
DPPH solution and tested for Antioxidant activity.
The aqueous extract of guava showed prominent bleaching of purple colour of DPPH indicating
that it has Antioxidant activity.
Qualitative antioxidant activity shown by aqueous extract of Psidium guajava.

Before DPPH. After DPPH

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STUDY OF ANTIOXIDANT ACTIVITY BY DPPH

The antioxidant activity of aqueous extract of guava leaves was assessed on the basis of radical
scavenging effect of the stable 1,1- diphenyl 2- picrylhydrazyl (DPPH). The diluted working
solutions of the test plant extract was prepared in water. 0.02% DPPH was prepared in ethanol
2 ml of solution having increasing concentration of guava extract, and optical density was
measured at 517nm using colorimeter. A blank reading of DPPH was also recorded and %
inhibition was calculated using the formula given below.

Percentage (%) Inhibition of DPPH (%AA) = A-B × 100

A
Where, A = Blank O.D of DPPH
B = O.D of sample solution.
From this we can calculate IC50 Value of each sample.
IC50 = Max (%AA) – 50% ( Max - Min%AA )
Result and discussion:

Study on Quantitative Antioxidant activity of water extract of guajava leaves.

Table: Optical density and percent Antioxidant of Aqueous extract of Guava.
Concentration 0.1 0.2 0.3 0.4 0.5 0.6 0.7 0.8 0.9
in mg/ml
O.D of sample 0.60 0.56 0.53 0.49 0.44 0.40 0.36 0.32 0.30

% AA 30.23 34.88 38.37 43.02 48.83 53.48 58.13 62.79 65.11

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IC50 = Max (%AA) – 50 % ( Max – Min % AA)

= 65.11 – ½ ( 65.11 – 30.23)
IC50 = 47.67

Conclusion:
The results obtained for the antioxidant assay by DPPH for water extract of leaves Psidium
guajava plant were reported. The remarkable decrease in O.D value of the test plant samples
were observed from the graph, showed antioxidant activity. The IC50 value for water extract of
leaves of Psidium guajava was found to be 47.67mg/ml .

References
1) Bipul Biswas, Kimberly Rogers and Anand Yadav. Antimicrobial Activities of leaf
extracts of guava. Int J Microbial. 2013 : 746165. Doi : 10. 1155/ 2013/ 74165, PMCID:
PMC3817707, PMID : 24223039, Oct 2013.
2) Rika Hartati, HashifahI. Nadifan, Irda Fidrianny. Crystal Guava: Evaluation of in Vitro
antioxidant capacities and Phytochemical content. The Scientific World Journal
2020(3) : 1-7, doi : 10 . 1155/ 9413727, License. CC BY, 2020.
3) Ravi Narayan Venkatachalam, Kanchahlata Singh, ThankamaniMarar. Phytochemical
Screening and in Vitro Antioxidant activity of guava. Home / Archives/ Vol. 2, No. 1;
free radical and Antioxidants. ISSN : 2231 – 2536, 2012.
4) EziucheAmadikeUgbogu, ChibuikeIbe, Emmanuel Dike, Victor ChibuereUde,
Celestine NwabuEkweogu. Arabian Journal of Chemistry. Volume 15, Issue 5, 103759,
May 2022.
5) Larissa Takeda, Lucas Laurindo, ElenGuiguer, AnupamBishayee. Food Reviews
International. DOI : 10. 1080/ 87559129. 2021. 2023819, Feb 2022.
6) MaryanZahin et al. Antioxidant and antimutagenicpotential of Psidiumguajava leaf
extracts. Drug ChemToxical. Apr. 2017.
7) N. S. Sampath Kumar, NorizahMhdSarbon, Sandeep Singh Rana, Anjani Devi,
Chintagunta et al. Extraction of bioactive compounds from Psidiumguajava leaves and

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its utilization in preparation of Jellies. (2021) 11: 36, https: // doi. org/ 10. 1186/ 513568
– 021 – 01194 – 9, 2021.
8) R. Manikandan and Vijay Anand. Antioxidant activity of Psidiumguajava. Research
Journal of Pharmacy and Technology. 8(3): 339, 2015.
9) SumraNaseer, Shabbir Hussain, NaureenNaeem, Muhammad Pervaiz and
MadhihaRahman. Phytochemical and medicinal value of guava. Clinical Phytoscience
(Vol. 4, Issue 1) ,Dec.12, 2018.
10) N.K. Jain, Zahoor Ahmad Lone. Phytochemical screening of guava. International
journal of innovation in engineering research and management. ISSN: 2348- 4918,
Aug 2022.
11) Manoj Kumar, RyszardAmarowicz, VivekSaurabh,Surinder Singh, SushilChandan,
Mukesh k. Berwal. Foods. 2021, Apr; 10(4) : 752. DOI: 10.3390/foods 10040752,
PMCID: PMC8066327.
12) AnumitaBhadra, RattandeepSingh. Phytochemical properties of guava plants. Vol.
11(2023) : Ahead of print 1, DOI: https://doi,org //10.7770/ Safer –V11N1-art 2385.
13) Arun K. Verma, V. Rajkumar and ArunK. Das. Guava Powder as an Antioxidant
Dietary fibre in Sheep Meat Nuggets. Asia –Australian Journal of Animal Sciences.
26(6) : 886 – 895.Doi: 10.5713/ ajas. 2012.12671, PMCID : PMC4093245, PMID :
25049864, 2013.
14) Laily N., Kusumaningtyas R.W., Sukarti I., Rini M.R.D.K. The potency of
guava Psidium guajava (L.) leaves as a functional immunostimulatory
ingredient. Procedia Chem. 2015;14:301–307. doi: 10.1016/j.proche.2015.03.042.
15) Chen H.Y., Yen G.C. Antioxidant activity and free radical-scavenging capacity of
extracts from guava (Psidium guajava L.) leaves. Food Chem. 2007;101:686–694.
doi: 10.1016/j.foodchem.2006.02.047.
16) . Ashraf A., Sarfraz R.A., Rashid M.A., Mahmood A., Shahid M., Noor N. Chemical
composition, antioxidant, antitumor, anticancer and cytotoxic effects of Psidium
guajava leaf extracts. Pharm. Biol. 2016;54:1971–1981.
doi: 10.3109/13880209.2015.1137604.
17) . Jiang L., Lu J., Qin Y., Jiang W., Wang Y. Antitumor effect of guava leaves on lung
cancer: A network pharmacology study. Arab. J. Chem. 2020;13:7773–7797.
doi: 10.1016/j.arabjc.2020.09.010.
18) Manoj kumar, Maharish Tomar, Ryzard Amarowicz, Vivek saurabh, M S Naire,Chirag
Maheshwari, Minnu sasi, Uma Prajapati, Muzzafar Hasan, Surinder singh, Shushil
changan, R K Prajapat, Mukesh Berwal, warsha Satankar, Foods. 2021 Apr; 10(4): 752.

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74

In-Silico Prediction of Phytoconstituents From Solanum Indicum for

Antiepileptic Activity

Pooja.P.Patle*, Parimal Katolkar, Pradeep Raghatate, Jagdish Baheti

Kamla Nehru College of Pharmacy, Butibori, Nagpur 441108 (MS), INDIA
*Email: [email protected]

ABSTRACT
Objective
A persistent, non-communicable brain disorder is epilepsy. Its distinctive characteristic is
recurrent seizures. One or more parts of the body may experience partial or generalised
seizures, which are short bursts of uncontrollable movement that can occasionally be followed
by loss of consciousness and control over bowel or bladder function. Compounds found in
medicinal plants have been the source of many conventional medications. In-silico testing of
Solanum indicum phytoconstituents for antiepileptic efficacy was a part of our investigation.
Methods
Utilizing Discovery studio, molecular docking is done to assess the pattern of interaction
between the phytoconstituents from the Solanum indicum plant and the crystal structure of the
epilepsy proteins (PDB ID: 6O4L). Later, SwissADME and pkCSM were used to screen for
toxicity as well as the pharmacokinetic profile.
Results
The docked results suggest that Solafuranone (-7.8 kcal/mol), Isofraxidin (-6.1 kcal/mol) for
6O4L macromolecule has best binding towards antiepileptic activity as compared to the
standard (Acetazolamide) for 6O4L is -5.1 kcal/mol. Furthermore, pharmacokinetics and
toxicity parameters were within acceptable limits according to ADMET studies.
Conclusion
Results from the binding potential of phytoconstituents aimed at antiepileptic activity were
encouraging. It promotes the usage of Solanum indicum and offers crucial details on
pharmaceutical research and clinical care.
1. INTRODUCTION
Solanum indicum, also known as Birhata, Badi Kateri, or Indian night shade, belongs to the
Solanaceae family. It is a common upright undershrub in warmer parts of India, Asia, and
Africa that grows up to 1.5 metres in height. Its height ranges from 0.30 to 1.8 metres.
Nationally, 500–1000 MT are needed each year. The berries, leaves, roots, seeds, and stems of
this plant have all been used in traditional medical systems to treat a wide range of ailments,
including bronchitis, asthma, dry cough, rhinitis, dysuria, leucoderma, sexual dysfunction,
insomnia, heart weakness, and pruritis.1
Solanum indicum, which is consumed as a vegetable in some regions of Africa, may prevent
cardiovascular diseases. It was exciting to read the studies on how it may be used to treat
hypertension. The extract's potent therapeutic and preventative benefits against hypertension
may not have been possible without the presence of chlorogenic acids. The antihypertensive
effect might be due to other components. The findings urge further research into the extract as
a possible treatment for hypertension.2
However, there are few studies on the phytoconstituents of Solanum indicum for the
antiepileptic activity. Thus, keeping the above information in view, the present investigation
was designed to identify the potential phytochemicals of Solanum indicum against 6O4L using
a molecular docking method.

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2. MATERIALS AND METHODS

2.1. Platform for molecular docking
The computational docking study of all the phytoconstituents selected as ligands with
antiepileptic activity as the target was performed using PyRx software.3
2.2. Protein preparation
The macromolecule is 6O4L, in silico analysis of selected phytoconstituents was performed on
the 1.85 Å crystal structure of antiepileptic macromolecule with inhibitor, (PDB ID: 6O4L,
having resolution Resolution: 1.85 Å, R-Value Free: 0.220, R-Value Work: 0.174, R-Value
Observed: 0.176), which was retrieved from the protein data bank (https://www.rcsb.org). 6O4L
is classified as Crystal Structure of ALDH7A1 mutant E399D complexed with NAD all other
molecules, such as co-crystallized water molecules, unwanted chains, and nonstandard
residues, were deleted. Using Discovery studio.4
2.3. Mechanism of Action
6O4L: Aldehyde dehydrogenase 7A1 (ALDH7A1) is an enzyme that catabolizes lysine, and
some loss-of-function mutations in this gene result in pyridoxine-dependent epilepsy (PDE).
To understand how the mutations affect NAD+ binding, the crystal structures of the mutant
enzymes in complex with NAD+ were examined. At the tested dose of NAD+, these
alterations result in less active tetrameric ALDH7A1.5
2.4. Ligand preparation
The three-dimensional (3D) structures of all constituents were retrieved using Avogadro
software from the PubChem database available on the NCBI website
(https://pubchem.ncbi.nlm.nih.gov/). However, the drawing of geometrical 2D structure was
performed using the ChemSketch program. The two-dimensional (2D) structures were
transformed into 3D models using the Avogadro software and the ligand structures were saved
in the PDB format. All the chemical structures are shown in Figure 1.

Solanidine Solasodine
OH H
H3C
H3C NH CH3
H3C H
H3C H CH3
HH O
H CH3
N H

HO
CH3

Solavetivone Solafuranone
CH2 H3C
CH3 H3C O
O
CH3
O
H3C CH3

CH3

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Scopoletin N-p-trans-Coumaroyltyramine

HO
H
N

O
OH

Isofraxidin Lauric acid

H3C O HO CH3

O
HO O O
O
H3C
𝛽-sitosterol Diosgenin
H
O
H3C

H3C H
H
H
O H

O H
H

Fig. 1. Chemical structures of all selected phytoconstituents in the molecular docking

studies
2.5. Standard Preparation
The standard is prepared steps such as, the 2D structure of standard drug was made using chem
sketch program, then the 2D structure was converted into 3D model using Avogadro Software,
it was saved in PDB format.
By using PyRx molecular docking of Acetazolamide was done with 6O4L.
2.6. Molecular docking
Molecular docking evaluates the protein-ligand interactions and estimates the scoring function
based on the geometry to predict the binding affinity of the ligand molecule6,7. We
applied molecular docking studies to investigate the binding pattern of selected
phytoconstituents (Figure 1) and the standard drug, along with the crystal structure of
antiepileptic activity macromolecule (PDB ID: 6O4L). The molecular docking study was
performed using PyRx software, Binding affinity was explored using the Vina wizard tool. The
final results were analysed and visualized using Discovery Studio 2020 Client 8, with bound
ligands as the standard. Visualization of protein ligand interaction reflects the number of
interactions and active residues responsible for significant binding at the active site of the target
enzyme.
2.7. Absorption, distribution, metabolism, and excretion (ADME) and toxicity prediction
The selected phytoconstituents and standard drug were further checked for drug-likeness
properties according to Lipinski’s rule. During drug development, it is necessary to predict the
tolerability of phytochemicals before being ingested by humans and animal models.

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The pharmacokinetic profile (ADME) and toxicity predictions of ligands were conducted using
SwissADME (http://www.swissadme.ch) and pkCSM (an online server database predicting
small-molecule pharmacokinetic properties using graph-based
signatures, (http://biosig.unimelb.edu.au/pkcsm/prediction). To analyse the toxicological
properties of ligands, Simplified Molecular Input Line Entry System (SMILES) notations or
PDB files were uploaded, followed by selecting the required models for generating numerous
information about structure-related effects9,10.
3. RESULT AND DISCUSSION
The present study aimed to explore the inhibitory potential of the phytoconstituents present
in Solanum indicum targeting antiepileptic activity. In this study, we performed molecular
docking studies of all phytoconstituents found in Solanum indicum using AutoDock Vina,
followed by a study of interacting amino acid residues and their influence on the inhibitory
potentials of the active constituents. Selected phytoconstituents showing the best fit were
further evaluated for absorption, distribution, metabolism, excretion, and toxicological
(ADMET) properties using SwissADME and pkCSM servers.
3.1 Molecular docking
The docking scores and binding energies of all chemical constituents of Solanum
indicum targeting antiepileptic activity (PDB ID: 6O4L) and binding interactions with amino
acid residues are presented in Table 1 respectively.

Table 1. Binding interaction of ligands from Solanum indicum targeting antiepileptic

activity (PDB ID:6O4L)

Chemical Docking Score

Sr. No. PubChem ID
constituent 6O4L
1. Solanidine 65727 -10.0
2. Solasodine 5250 -9.0
3. Solavetivone 442399 -7.3
4. Solafuranone 11107208 -7.8
5. Scopoletin 5280460 -6.5
N-p-trans-
6. 5372945 -6.7
Coumaroyltyramine
7. Isofraxidin 5318565 -6.1
8. Lauric acid 3893 -3.7
9. 𝛽-sitosterol 222284 -7.1
10. Diosgenin 99474 -8.1
Standard Drug
11 Acetazolamide 1986 -5.1

The binding affinities of phytoconstituents ranged from –10.0 to -3.7 kcal/mol. From the
docked results, it is evident that the compounds, Solanidine exhibit the most favourable binding
affinity (−10.0 kcal/mol) in complex with selected macromolecules (PDB ID: 6O4L) as
compared to other docked compounds i.e., Solsodine (-9.0 kcal/mol), Diosgenin (-8.1
kcal/mol), Solafuranone (−7.8. kcal/mol), Solavetivone (−7.3 kcal/mol), Beta sitosterol (-7.1
kcal/mol), N-p-trans-Coumaroyltyramine (−6.7 kcal/mol), Scopoletin (−6.5 kcal/mol),
Isofraxidin (−6.1 kcal/mol), Lauric acid (−3.7 kcal/mol). Visual examination of the
computationally docked optimal binding poses of phytoconstituents on selected
macromolecules (i.e., 6O4L) revealed the significant involvement of various types of
interactions, such as hydrogen bonding and hydrophobic interactions, including π–π stacking

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and π–alkyl and alkyl interactions, in the stability of the binding of the phytoconstituents to
6O4L.
The binding affinity of the standard (Acetazolamide) for 6O4L is -5.1 kcal/mol.
3.1.1. Solafuranone, 6O4L
The number of intermolecular hydrogen bonds, the binding energy of ligand 6O4L stable
complexes, and the number of nearest amino acid residues were also determined for selected
compound Solafuranone All synthesized derivatives formed complexes with target proteins.
Analysis of interactions of the 6O4L protein complex and ligand Solafuranone showed that the
ligand molecule is oriented due to Conventional Hydrogen bond with THR A: 346 amino acid
residue and Pi-Pi Stacked interaction with PHE A: 401 amino acid residue were found.
3.1.2. Isofraxidin, 6O4L
An analysis of the interactions between the 6O4L protein complex and the Isofraxidin ligand
was also carried out, which showed that the ligand molecule is oriented due to conventional
hydrogen bond with the ASP A: 399, THR A: 346, PHE A: 401 amino acid residue and Pi-
Alkyl interaction with ALA A: 349 amino acid residue were found.
3.1.3. Acetazolamide, 6O4L
The binding affinity of the standard (Acetazolamide) for 6O4L is -5.1 kcal/mol. the interactions
between the 6O4L protein complex and the Acetazolamide ligand was also carried out, which
showed that the ligand molecule is oriented due to conventional hydrogen bond with the PHE
A:166, THR A:164 amino acid residue, Carbon hydrogen bond with PRO A:193 amino acid
residue, Pi-Sulfur interaction with PHE A:166 amino acid residue, Pi-Cation interaction with
LYS A:190 and Pi-Alkyl interaction with ALA A:165, ALA A:192 amino acid residue were
found.
Table No. 2. Interactions with amino acid residue.
Main amino acid interactions
Binding
Sr. Pi-alkyl, alkyl, Pi-S/Pi-Pi, T Van der
Molecule Energy H bond
No. shaped/halogen/unfavourable Waals
(kcal/mol)
donor-donor interactions interactions
No ALA A:192, ALA A: 165, No
1 Solanidine -10.0
interactions VAL A: 250 interactions
GLY A: No
2 Solasodine -9.0 VAL A: 250, PRO A: 193
226 interactions
ASN A:
No
3 Solavetivone -7.3 379, ASN TYR A: 41
interactions
A: 46
THR A: No
4 Solafuranone -7.8 PHE A: 401
346 interactions
THR A:
No
5 Scopoletin -6.5 164, PHE CYS A: 302, ALA A: 165
interactions
A: 401
N-p-trans- SER A: PHE A: 168, CYS A: 302, TRP No
6 -6.7
Coumaroyltyramine 460 A: 175, PHE A: 468 interactions
ASP A: No
7 Isofraxidin -6.1 ALA A: 349
399, PHE interactions
PHE A: PRO A: 288, LEU A: 285, PRO No
8 Lauric acid -3.7
292 A: 458 interactions
TRP A: 31, ALA A: 207, LYS
No No
9 𝛽-sitosterol -7.1 A: 204, LYS A: 208, VAL A:
interactions interactions
209, ALA A: 92
ASN A: No
10 Diosgenin -8.1 TYR A: 41, PHE A: 166
379 interactions

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THR A:
164, PHE
ALA A: 165, ALA A: 192, No
11 Acetazolamide -5.1 A: 166,
LYS A: 190 interactions
PRO A:
193

3.2. ADMET study

Pharmacokinetic profile (ADME) and toxicity predictions of the ligands are important attentive
parameters during the transformation of a molecule into a potent drug. In the present study,
these parameters were assessed using SwissADME and pkCSM. The absorption potential and
lipophilicity are characterized by the partition coefficient (Log P) and topological polar surface
area (TPSA), respectively. For better penetration of a drug molecule into a cell membrane, the
TPSA should be less than 140 Å. However, the value of Log P differs based on the drug target.
The ideal Log P value for various drugs are as follows: oral and intestinal absorption, 1.35 −
1.80; sublingual absorption, > 5; and central nervous system (CNS)11. The aqueous solubility of
ligands ideally ranges from − 6.5 to 0.5 12, while the blood brain barrier (BBB) value ranges
between − 3.0 and 1.2 13. In addition, non-substrate P-glycoprotein causes drug resistance 14.
In our study, all the selected ligands followed the TPSA parameter, P-glycoprotein non-
inhibition, thereby showing good intestinal absorption and an acceptable range of BBB
values. All the compounds showed aqueous solubility values within the range. Further, it was
predicted that the selected ligands do not show AMES toxicity, hepatotoxicity, and skin
sensitivity. In addition, it did not inhibit hERG-I (low risk of cardiac toxicity). Lipinski’s rule
violations, T. pyriformis toxicity, minnow toxicity, maximum tolerated dose, rat acute oral
toxicity, and chronic toxicity are depicted in table.

Table 3. ADME and toxicity predicted profile of ligands with superior docking score
Aqueou
Human Blood
MW HB Hb s
ADMET TPSA intestinal Brain
Formula (g/mol Log P done accepto solubilit
Properties (Ȧ2) absorptio Barrie
) r r y (Log
n (%) r
mol/L)
397.64 23.47
Solanidine C27H43NO 5.655 1 2 -4.927 92.975 0.695
7 Ȧ2
41.49Ȧ
Solasodine C7H43NO2 413.64 5.2869 2 2 3 -3.809 92.324 0.035
17.07Ȧ
Solavetivone C15H22O 218.33 3.9042 2 0 1 -4.615 95.873 0.635
232.32 3.1876 26.30Ȧ
Solafuranone C15H20O2 2 0 2 -3.551 95.523 0.206
3 4
59.67Ȧ
Scopoletin C10H8O4 192.17 1.5072 2 1 4 -2.504 95.277 -0.299
N-p-trans-
283.32 69.56Ȧ
Coumaroyltyrami C17H17NO3 2.4699 2 3 3 -3.165 90.031 -0.552
7
ne
222.19 68.90Ȧ
Isofraxidin C11H10O5 1.5158 2 1 5 -2.458 95.588 -0.377
6
200.32 37.30Ȧ
Lauric acid C12H24O2 3.9919 2 1 1 -4.181 93.379 0.057
2
414.71 20.23Ȧ
𝛽-sitosterol C29H50O 8.0248 2 1 1 -6.773 94.464 0.781
8
38.69Ȧ
Diosgenin C27H42O3 414.63 5.7139 2 1 3 -5.713 96.565 0.2
C4H6N4O3S 222.25 - 151.66
Acetazolamide 2 6 -2.428 59.043 -0.622
2 1 0.8561 Ȧ2

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Table 3 Continued
Total Max
clearanc tolerate
P-glyco- AME hERG
e Bioavailabilit d hERG I
ADMET protein S II
[Log y score dose inhibito
Properties substrat toxicit inhibito
mL/ score [Log r
e y r
(min.kg mg/
)] (kg.d)]
Solanidine YES 0.028 0.55 NO -0.882 NO YES
Solasodine YES 0.09 0.55 NO -0.375 NO YES
Solavetivone NO 1.225 0.55 NO 0.044 NO NO
Solafuranone NO 1.256 0.55 NO 0.526 NO NO
Scopoletin NO 0.73 0.55 NO 0.614 NO NO
N-p-trans-
Coumaroyltyrami YES 0.265 0.55 NO -0.213 NO YES
ne
Isofraxidin NO 0.713 0.55 NO 0.56 NO NO
Lauric acid NO 1.623 0.85 NO -0.34 NO NO
𝛽-sitosterol NO 0.628 0.55 NO -0.621 NO YES
Diosgenin NO 0.328 0.55 NO -0.559 NO YES
Acetazolamide NO -0.01 0.55 NO 1.263 NO NO

Table 3 Continued
Oral rat Minno
T.Pyrifor
Acute oral chronic w Lipinski
Skin mis
ADMET rat toxicity. toxicity Hepatotoxic toxicit ’s rule
sensati toxicity
Properties LD50(mol/ (Log ity y (Log Violatio
on (Log
kg) mg/kgbw/d mmol/ n
μg/L)
ay) L)
Solanidine 2.596 1.334 YES NO 0.378 -0.493 YES (1)
Solasodine 2.489 1.332 YES NO 0.311 0.381 YES (1)
Solavetivone 1.643 1.19 NO YES 1.453 0.874 YES (0)
Solafuranone 1.865 1.947 NO YES 2.151 0.557 YES (0)
Scopoletin 1.95 1.378 NO NO 0.516 1.614 YES (0)
N-p-trans-
Coumaroyltyra 2.17 1.271 NO NO 1.008 1.514 YES (0)
mine
Isofraxidin 2.326 1.825 NO NO 0.431 1.862 YES (0)
Lauric acid 1.511 2.89 NO YES 0.954 -0.084 YES (0)
𝛽-sitosterol 2.552 0.855 NO NO 0.43 -1.802 YES (1)
Diosgenin 1.921 1.452 NO NO 0.399 0.247 YES (1)
Acetazolamide 2.292 2.143 NO NO 0.239 2.895 YES (0)

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3.3. Interaction of Standard Drug (Acetazolamide) with 6O4L

Fig. 2. Docking scores and binding interaction of Acetazolamide (PDB ID: 6O4L). The ligand
is shown in line and stick representation along with its 2D diagram and hydrogen bond
interaction.

3.4. Interactions of phytoconstituents with 6O4L

A.Solafuranone

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B.Isofraxidin

Fig. 3. Docking scores and binding interaction of phytoconstituents (PDB ID: 6O4L). The
ligand is shown in line and stick representation along with its 2D diagram and hydrogen bond
interaction.

3.5. Boiled Egg

Fig no. 4. Combined Boiled Egg Diagram

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Table no. 4. Name of molecules contained in Boiled Egg Diagram

MOLECULE NUMBER MOLECULE NAME
1 Solanidine
2 Solasodine
3 Solavetivone
4 Solafuranone
5 Scopoletin
6 N-p-trans-Coumaroyltyramine
7 Isofraxidin
8 Lauric acid
9 𝛽-sitosterol
10 Diosgenin
11 Acetazolammide

BOILED means Brain Or IntestinaL EstimateD permeation predictive model. The boiled egg diagram
shows two regions white region and yellow region.
The white region is the physicochemical space of molecules with highest probability of being absorbed
by the gastrointestinal tract, and the yellow region (yolk) is the physicochemical space of molecules
with highest probability to permeate to the brain.
In addition, the points are coloured in blue if predicted as actively effluxed by P-gp (PGP+) and in red
if predicted as non-substrate of P-gp (PGP-).

4. CONCLUSION
In this study, we have carried out an in-silico screening of the phytoconstituents of Solanum
indicum plant. This study demonstrated the sixteen compounds from Solanum indicum plant,
(Solanidine, Solasodine, Solavetivone, Solafuranone, Scopoletin, N-p-trans-
Coumaroyltyramine, Isofraxidin, Lauric acid, 𝛽-sitosterol, Diosgenin). The selected
phytocompounds showed docking scores ranging from –10.0 to −3.7 kcal/mol in 6O4L.
Among all, Solafuranone and Isofraxidin gave the highest binding energy (−7.8 kcal/mol) and
(−6.1 kcal/mol) in complex with 6O4L, whereas the reference compound, Acetazolamide
showed a docking score with a binding energy of -5.1 kcal/mol. Furthermore, these ligands
exhibited good ADMET properties. To summarize, phytoconstituents present in Solanum
indicum possess strong effects against 6O4L and could be further evaluated for their
antiepileptic effect, as well as for the development of alternative drugs with fewer side effects
for the treatment of epilepsy.

REFRENCES
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HPTLC fingerprinting of different parts of Solanum indicum L.: A dashmool species.
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2. Bahgat A, Abdel‐Aziz H, Raafat M, Mahdy A, El‐Khatib AS, Ismail A, Khayyal MT.
Solanum indicum ssp. distichum extract is effective against l‐NAME‐induced hypertension
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5. Liles JT, Corkey BK, Notte GT, Budas GR, Lansdon EB, Hinojosa-Kirschenbaum F, Badal
SS, Lee M, Schultz BE, Wise S, Pendem S. ASK1 contributes to fibrosis and dysfunction
in models of kidney disease. The Journal of Clinical Investigation. 2018 Oct
1;128(10):4485-500.
6. Verdonk ML, Cole JC, Hartshorn MJ, Murray CW, Taylor RD. Improved protein–ligand
docking using GOLD. Proteins: Structure, Function, and Bioinformatics. 2003
Sep;52(4):609-23.
7. Leach AR, Shoichet BK, Peishoff CE. Prediction of protein− ligand interactions. Docking
and scoring: successes and gaps. Journal of medicinal chemistry. 2006 Oct 5;49(20):5851-
5.
8. SAMANT L, Javle V. Comparative docking analysis of rational drugs against COVID-19
main protease.
9. Arora S, Lohiya G, Moharir K, Shah S, Yende S. Identification of potential flavonoid
inhibitors of the SARS-CoV-2 main protease 6YNQ: a molecular docking study. Digital
Chinese Medicine. 2020 Dec 1;3(4):239-48.
10. Shah S, Chaple D, Arora S, Yende S, Moharir K, Lohiya G. Exploring the active
constituents of Oroxylum indicum in intervention of novel coronavirus (COVID-19) based
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11. Kaloni D, Chakraborty D, Tiwari A, Biswas S. In silico studies on the phytochemical
components of Murraya koenigii targeting TNF-α in rheumatoid arthritis. Journal of Herbal
Medicine. 2020 Dec 1;24:100396.
12. Joshi T, Sharma P, Joshi T, Chandra S. In silico screening of anti-inflammatory compounds
from Lichen by targeting cyclooxygenase-2. Journal of Biomolecular Structure and
Dynamics. 2020 Aug 12;38(12):3544-62.
13. Nisha CM, Kumar A, Vimal A, Bai BM, Pal D, Kumar A. Docking and ADMET prediction
of few GSK-3 inhibitors divulges 6-bromoindirubin-3-oxime as a potential inhibitor.
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in patients with rheumatoid arthritis. World journal of experimental medicine. 2015 Nov
11;5(4):225.

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75

Development and Characterization of Piroxicam Matrix Based

Transdermal Patch

Pragati Hasbe , Zohra Firdous M.S.A*, Pankaj Dhapke, Jagdish Baheti

Kamla Nehru College of Pharmacy, Butibori, Nagpur 441108, Maharashtra, India.
e-mail address: [email protected]

ABSTRACT:
The present study aims to develop and characterize matrix based transdermal patch for the
treatment of dysmenorrhea using the drug Piroxicam, which is a non-steroidal anti-
inflammatory drug of oxicam derivative. Matrix-based transdermal patches were prepared
using a solvent casting method consisting of polymer (HPMC E5LV) and drug along with PEG
400 (plasticizer) and capsicum oleoresin (penetration enhancer) in a methanol and
dichloromethane solution. All F8 formulation batches prepared were evaluated and optimized.
The result shows that the F3 batch gives consistent results and all the evaluated parameters were
within the standard limits. The in-vitro and ex-vivo %CDR of the optimized F3 batch was found
to be 98.76% and 97.65% respectively which indicates that the permeability of the drug through
the goat skin within the time period of 2 hours is less than that of the dialysis membrane.The
research work concluded that piroxicam matrix based transdermal patch was successfully
prepared and evaluated which can provide direct entry of the drug into systemic circulation for
immediate action in 2 hours. Therefore, piroxicam matrix based transdermal patches can be
used to safely deliver the drug for the treatment of dysmenorrhea while avoiding therapy
associated side effects and with good patient compliance.

KEYWORDS: Dysmenorrhea, Piroxicam matrix based transdermal patch, Capsicum

oleoresin, HPMC E5LV, PEG 400.

INTRODUCTION
Dysmenorrhea has a negative impact on social, academic, sporting and daily activities. It is the
most common problem that can be seen in adolescent and young women all over the world.
Worldwide statistics shows that 51% of adolescent girls suffer from dysmenorrhea.
Dysmenorrhea can be treated by using non-steroidal anti-inflammatory drugs (NSAIDs).
Piroxicam is a non-steroidal anti-inflammatory drug of the oxicam derivative that are used to
relieve pain, stiffness, tenderness and swelling by preventing the production of endogenous
prostaglandins. This results in inhibition of prostaglandin biosynthesis. It has a role as an
analgesic, a COX1 inhibitor, a non-steroidal anti-inflammatory drug, antipyretic, etc. It is used
in the treatment of certain inflammatory conditions like primary & secondary dysmenorrhea,
rheumatoid and osteoarthritis and postoperative pain. The transdermal drug delivery system
(TDDS) is a route of administration where in active ingredients are delivered across the skin
for systemic distribution. Transdermal patch is one of the conventional forms of transdermal
drug delivery system.TDD is a painless method of delivering drugs systemically by applying a
drug formulation onto intact and healthy skin. The drug initially penetrates through the stratum
corneum and then passes through the deeper epidermis and dermis without drug accumulation
in the dermal layer. When drug reaches the dermal layer, it becomes available for systemic
absorption via the dermal microcirculation. Therefore, this work is formulated and evaluated
the piroxicam matrix based transdermal patch for the treatment of Dysmenorrhea.

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Application of Transdermal Delivery System

 Transdermal patches adhere to the skin as a way to deliver drugs. They provide a specific,
predetermined dose of medication that is absorbed through the skin and penetrates into the
bloodstream
 NSAIDs had been in first line of choice for transdermal drug delivery system for a long
times due to their improved local effects and possibility to avoid the gastro-irritating effects.
 NSAIDs transdermal patches are most suitable techniques for reducing the adverse effects,
increasing the bioavailability by avoiding first-pass hepatic metabolism, elevating the
permeability of NSAIDs, and facilitating sustained release of drug for the longer duration
of time.
 The drugs such as analgesic, antipyretic, antidepressant, antianginal and NSAIDs can be
given by using transdermal drug delivery system.
MATERIALS AND METHODS
Chemicals
Piroxicam was purchased from Yarrow Chem Products, Mumbai, India. HPMC E5 LV was
procured from Central Scientific Company, Nagpur, India. PEG 400 and methanol was
procured from The Global Marketing, Nagpur, India and dichloromethane were procured from
Genex Scientific Company, Nagpur, India. Capsicum oleoresin was purchased from Avi
Naturals, Delhi, India. All other chemicals and reagents used in the study were of analytical
grade.
Method of preparation of matrix based transdermal patch

RESULTS AND DISCUSSION

Evaluation of Transdermal patches
Physical appearance
The formulated patches were found to be clear, smooth, uniform, flexible and light yellow
colour in their physical appearance as shown in figure

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In-vitro diffusion cell study

% %
Drug
Weight Folding Moisture Moisture
Formulation Thickness Content
Uniformity Endurance Content Uptake
Code (mm)±S.D (%) ±
(mg) ± S.D ± S.D (%w/w) (%w/w)
S.D
± S.D ± S.D
0.690 ± 540.75 ± 2.84 ± 98.8 ±
F1 7.67 ± 1.16 6.6 ± 0.19
0.051 0.88 0.57 0.45
0.554 ± 499.5 ± 2.53 ± 4.25 ± 98.4 ±
F2 6±1
0.024 0.75 0.21 diffusion0.51
0.25 Figure 1: In-Vitro cell study for % Cumulative
0.323 ± 301.12 ± 2.24 ± 3.31 ± Drug 99.17 ±
Release
F3 9±1
0.007 0.35 0.02 0.13 0.19
0.648 ± 4.14 ± 98.18 ±
F4 566 ± 0.92 5.67 ± 0.58 5.2 ± 0.56
0.048 0.31 0.33
0.544 ± 470.37 ± 2.64 ± 94.1 ±
F5 6.33 ± 2.31 4.6 ± 0.29
0.018 0.74 0.19 0.70
0.615 ± 520.5 ± 4.36 ± 95.8 ±
F6 8±2 5.95±0.37
0.036 0.80 0.04 0.39
0.337 ± 355.75 ± 2.45 ± 3.77 ± 97.8 ±
F7 6.66 ± 1.53
0.008 0.46 0.03 0.27 0.79
0.450 ± 388.62 ± 3.03 ± 5.88 ± 98.6 ±
F8 7.33 ± 1.52
0.009 0.51 0.06 0.20 0.25
The % of drug release orders was as follows: F3 > F1 > F8 > F2 > F4 > F7 > F6 > F5. The
formulation F3 batch showed a better in-vitro drug release profile across the dialysis membrane
when compared to the other formulations (figure 12). This might be attributed to the nature of
the polymer; plasticizer and the permeation enhancer used. Thus formulation F3 is considered
an optimized formulation
Ex-vivo skin permeation study
The F3 batch ex-vivo skin permeation study was performed. The patch was exposure to the
goat skin, 97.65 % of the drug was permeated and 98.76 % of the drug was permeated from in-
vitro diffusion of optimized F3 batch as shown in table 12. Thus, the amount of the drug
permeated through the goat skin was less than the dialysis membrane. The graph (figure 13) of
cumulative drug release, ex-vivo skin permeation and in-vitro cell study of an optimized batch
(F3)

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Figure 2: % Cumulative Drug Release ex-vivo skin permeation Vs in-vitro study of an

optimized batch (F3)

Sr. Time In-Vitro %CDR Ex-Vivo %CDR (F3) Stability study of optimized F3
No. (min.) (F3) (%) (%) batch
1 0 0 0 Stability study was performed
2 20 8.52 ± 1.16 4.25 ± 1.39 for optimized F3 batch
(piroxicam matrix based
3 40 27.46 ± 1.75 15.12 ± 2.26 transdermal patch) was
4 60 44.19 ± 3.47 34.69 ± 3.96 wrapped in aluminium foil by
placing in the zip lock bag and
5 80 65.8 ± 5.34 57.40 ± 1.94
kept in stability chambers at
6 100 81.85 ± 5.29 80.06 ± 1.95 40°C/75% RH for accelerated
7 120 98.76 ± 0.57 97.65 ± 0.65 study of 6 months. Formulation
was removed at each time point
(0 day, 1 month, 3 month and 6 month) and evaluated for physical appearance, weight
uniformility, thickness, folding endurance and drug contents. The in-vitro & ex-vivo
permeation study were also performed. The stability study was completed as shown in table
In- Ex-
Weight Drug Vitro Vivo
Time Thickness Folding
Appearance Uniformity Content %CDR %CDR
(months) (mm) Endurance
(mg) (%) (F3) (F3)
Pale light 0.323 ± 301.12 ± 99.17 ± 98.76 ± 97.65 ±
0 9±1
yellow 0.007 0.35 0.19 0.57 0.65
Pale light 0.320 ± 98.95 ± 98.35 96.76 ±
1 298 ± 0.39 10 ± 1.15
yellow 0.008 0.35 ± 0.71 0.61
Pale light 0.324 ± 98.43 ± 98.12 95.98 ±
3 306.1 ± 0.1 12 ± 2.2
yellow 0.010 0.5 ± 1.6 1.5
Pale light 0.319 ± 299.07 ± 98.54
6 10 ± 1.05 99 ± 1 97 ± 1
yellow 0.009 0.37 ± 0.8

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CONCLUSION
It is concluded that piroxicam matrix based transdermal patch was successfully formulated and
evaluated which may provides direct entry of the drug into the systemic circulation for
immediate action in 2 hours. The identification of drug shows pure piroxicam. The prepared
matrix based transdermal patch evaluation reveals that the F3 batch is an optimized batch which
shows good uniformity in the evaluation parameters of the patch. Such drugs delivery systems
can be used to avoid side effects associated with therapy and safely deliver the drug for the
treatment of dysmenorrhea with good patient compliance.

References
1. Azagew AW, Kassie DG, Walle TA. Prevalence of primary dysmenorrhea, its
intensity, impact and associated factors among female students’ at Gondar town
preparatory school, Northwest Ethiopia. BMC women's health. 2020;20(1):1-7
2. Singh A, Kiran D, Singh H, Nel B, Singh P, Tiwari P. Prevalence and severity of
dysmenorrhea: a problem related to menstruation, among first and second year female
medical students. Indian J Physiol Pharmacol. 2008;52(4):389-97.
3. Dharshini AV, Sangeetha A, Hemachandrika C. Primary dysmenorrhea and its impact
on academic performance among adolescent females–a cross sectional study. Annals
of the Romanian Society for Cell Biology. 2021:13681
4. Negriff S, Dorn LD, Hillman JB, Huang B. The measurement of menstrual symptoms:
factor structure of the menstrual symptom questionnaire in adolescent girls. Journal of
health psychology. 2009;14(7):899-908.
5. Rafique N, Al‐Sheikh MH. Prevalence of primary dysmenorrhea and its relationship
with body mass index. Journal of Obstetrics and Gynaecology Research.
2018;44(9):1773-8.
6. Speer LM, Mushkbar S, Erbele T. Chronic pelvic pain in women. Am Fam Phys.
2016;93:380.
7. Kulkarni A, Deb S. Dysmenorrhoea obstetrics, gynecology & reproductive medicine.
Elsevier. 2019; 29(10):286-291.
8. Ju H, Jones M, Mishra G. The prevalence and risk factors of dysmenorrhea.
Epidemiologic reviews. 2014 Jan 1;36(1):104-13.
9. Osayande AS, Mehulic S. Diagnosis and initial management of dysmenorrhea. Am
Fam Physician. 2014;89(5):341-6.
10. Sultan C, Paris F, Jeandel C, Lumbroso S, Galifer RB, Picaud JC. Ambiguous genitalia
in the newborn: diagnosis, etiology and sex assignment. Pediatric and Adolescent
Gynecology. 2004; 7:23-38.

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76

Analytical Method Development and Validation of Antidiabetic Drugs

Prajakta Sontakke, Disha Dhabarde, Jagdish Baheti
Kamla Nehru College of Pharmacy, Butibori, Nagpur
[email protected]
[email protected]

ABSTRACT
The present study focuses on the analytical method development and validation of
antidiabetic drugs by Uv-visible spectrophotometry and High-Performance Liquid
Chromatography. Developing single analytical method is challenging task. A simple, rapid,
precise and reliable method was developed for the estimation of bulk antidiabetic drugs. The
estimation was carried out using an Inertsil-C18 BDS column (250 mm × 4.6 mm, 5 µm) and
the mobile phase composed of methanol: H2O (80:20) at flow rate 1.0ml/min. Detection and
Quantification were performed by Uv-visible detection at 230 nm. The retention time of
metformin and dapagliflozin was found to be 3.08 and 3.87 minutes respectively. The validated
method was successfully applied to the commercially available pharmaceutical dosage form,
yielding good and reproducible results.

Keywords: Metformin and Dapagliflozin, Uv-visible spectrophotometry and High-

Performance Liquid Chromatography

INTRODUCTION:
Metformin is a biguanide antihyperglycemic agent and first-line
pharmacotherapy used in the management of Type II diabetes.
Metformin is considered an antihyperglycemic drug because it lowers
blood glucose concentrations in Type II diabetes without causing
hypoglycemia. It is commonly described as an "insulin sensitizer",
leading to a decrease in insulin resistance and a clinically significant
reduction of plasma fasting insulin levels. Fig.1
Dapagliflozin is a sodium-glucose cotransporter 2 (SGLT2) inhibitor,
and it was the first SLGT2 inhibitor to be approved. indicated for
managing diabetes mellitus type 2. When combined with diet and
exercise in adults, dapagliflozin helps to improve glycemic control by
inhibiting glucose reabsorption in the proximal tubule of the nephron
and causing glycosuria.
Fig2
Method
Selection of wavelength
Wavelength determination for HPLC analysis involved recording UV spectra (200-400 nm)
for individual Metformin and Dapagliflozin solutions. UV spectrum of standard Metformin and
Dapagliflozin was observed at 230 nm.
Chromatographic conditions
The developed method employs a reverse-phase C18 column, with a mobile phase consisting
of Methanol: H2O in the ratio of 80:20 v/v. The mobile phase flows at a rate of 1.0 ml/min,
and each injection involves 20μl of the sample. Detection was performed at a wavelength of
230 nm.
Preparation of Standard drug Solution of Metformin and Dapagliflozin

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Accurately weighed quantity of 120 mg Metformin diluted to 10 ml of diluent and 24 mg of

Dapagliflozin was dissolved in diluent and volume was made up to 100 ml mark by same to
obtain 1000 µg/ml. Then further diluted to obtain 100 ppm stock solution. From standard stock
solution pipette out 2 ml and dissolved in 20 ml of diluent to obtain 10 µg/ml of solution.
RESULT AND DISCUSSION
Method development
A reverse-phase HPLC method was developed with consideration for key system suitability
parameters, including the resolution factor (Rs) between peaks, peak asymmetry (A), number
of theoretical plates (N), run time, and cost-effectiveness. In conclusion, the developed method
is deemed specific. System precision was evaluated through six replicate injections of the
standards mixture at working concentration, revealing a % RSD (Relative Standard Deviation)
less than 2 for peak area of both drugs. This signifies acceptable reproducibility and precision
within the system.

Fig4
Method validation
The HPLC method developed underwent validation following International Conference on
Harmonization (ICH) guidelines. The validation included assessments for linearity, accuracy,
system precision, intra-day precision, ruggedness, limit of detection (LOD), and limit of
quantification (LOQ). Specificity was confirmed by observing peaks only in the standards and
sample solutions at working concentrations, with no interference from the blank. The method
is considered specific.
System precision was evaluated through six replicate injections of the standards solution at
working concentration, showing % RSD (Relative Standard Deviation) less than 2 for peak
area of both drugs, indicating acceptable reproducibility and precision.
Method precision was determined by assaying the sample under repeatability conditions
(intraday precision) at working concentrations.
System suitability
System suitability is an essential pharmacopeial requirement utilized to assess the
reproducibility of a chromatographic system, ensuring its adequacy for accurate analysis
In the study was assessed through recovery studies, involving the addition of standard drug
solution to reanalyzed sample solutions at three concentrations: 80%, 100%, and 120% of the
spiked levels. The accuracy was determined based on recovery studies conducted using the
spiked method, covering the range of 80-120% of the labeled claim.
Linearity
Linearity in analytical methods indicates results directly proportional or following a defined
mathematical transformation relative to analyte concentration within a specified range.
Assessment involves analyzing samples across the claimed concentration range, creating a
graph of area versus analyte concentration, and calculating percentage curve fittings to confirm
linearity

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Precision
Acceptance criteria require that the Relative Standard Deviation (RSD) should not exceed 2%
for the test. The Standard Deviation (SD) and the Relative Standard Deviation (RSD) are
calculated from the mean of five injections of the Standard solution and reported for evaluation.

Robustness
Robustness testing of the analytical method aimed to verify its stability against minor variations
in optimized parameters. Small changes were introduced in factors such as flow rate, mobile
phase composition, and wavelength. Specifically, variations of ±0.1 ml/min in flow rate, ±5 in
mobile phase composition, and ±2 nm in wavelength were implemented. The robustness of the
method was assessed by calculating the % RSD values to ensure the method's reliability despite
these slight variations.
SUMMARY AND CONCLUSION
An RP-HPLC method
Name Preparation %Assay was established and
applied to pharmaceutical
dosage forms for
Preparation-1 100.27
1.0 ml/mi Metformin and
n Dapagliflozin. A
Preparation-2 98.86 validated simple reverse-
phase liquid
Preparation-1 98.95 chromatographic method
0.9 ml/mi
was developed. A UV-
n
Preparation-2 99.08 spectrophotometric
method has been
Mean 99.29 developed,
demonstrating accurate
SD 0.6595 results for estimating
drugs in a mixture. The
%RSD (NMT 2) 0.66 separation method
utilized a mobile phase
composed of Methanol, Water, and Trifluoroacetic acid in a ratio of 80:20. Detection was
performed using UV–Visible SPD 20 A at 230 nm. The column employed was Hypersil BDS
C18 (250 mm × 4.6 mm, 5μm), and the flow rate was set at 1 ml/min.
The retention times for Metformin and Dapagliflozin were determined as 3.08 and 3.87,
respectively. The asymmetry factors or tailing factors of 1.17 and 1.10 suggested a symmetrical
peak nature at ambient temperature with an injection volume of 20 µl. The number of
theoretical plates was determined as 17196 and 15475, indicating the efficiency performance
of the column. Linearity studies were conducted, and specified concentration levels were
determined.
The method was validated according to ICH guidelines, demonstrating accuracy, precision,
selectivity, and cost-effectiveness. With sharp and well-defined peaks, the method is suitable
for the estimation of Metformin and Dapagliflozin in both bulk and pharmaceutical dosage
forms.

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REFERENCES
1. Chin B. and Anusha M., “Method Development And Validation For Symultaneous
Estimation Of Citicoline And Methylcobalamin By Rp-Hplc Method”, IPAPR, 2015,
6(10), 342-8.
2. Keval L. and Dilip G., “Rp-Hplc Method For The Estimation Of Epalrestat And
Methylcobalamin In Their Combined Dosage Form”, Indo American Journal of
Pharmaceutical Research, 2014, 4(6), 2697-2705
3. Chatwal A, "Instrumental Method of Chemical Analysis”, Himalaya Publishing House,
p.no.615-623.
4. Bhardwaj S., Dwivedi K, Agarwal D. A review: HPLC method development and
validation. International Journal of Analytical and Bio analytical Chemistry. 2015;
5(4):76-81.
5. Charde M., Welankiwar A.S, Kumar J. Method development by liquid chromatography
with validation. International Journal of Pharmaceutical Chemistry. 2014; 4(02):57-61.
6. Murugan S, Elayaraja A, Niranjan Babu M, Chandrakala K, Prathap Naik K, Ramaiah P,
Vulchi C. A Review on Method Development and Validation by using HPLC.
International journal of novel trends in pharmaceutical sciences. 2013 Oct; 3(3):78-81.
7. Hanif A, Bushra R, Ismail NE, Bano R, Abedin S, Alam S, Khan MA, Arif HM.
Empagliflozin: HPLC based analytical method development and application to
pharmaceutical raw material and dosage form. Pak. J. Pharm. Sci. 2021 May; 34(3):1081-
7.
8. Manoel J, Primieri GB, Bueno LM, Wingert NR, Volpato NM, Garcia CV, Schapoval EE,
Steppe M. The application of quality by design in the development of the liquid
chromatography method to determine Empagliflozin in the presence of its organic
impurities. RSC advances. 2020; 10(12):7313-20.
9. Shirisha V, Krishnaveni B, Illendula S, Rao KN, Dutt HR. A new simple method
development, validation and forced degradation studies of Empagliflozin by using Rp-
Hplc. International Journal of Pharmacy and Biological Sciences. 2019; 9(1):25-35.
10. 16. Naseef H, Moqadi R, Qurt M. Development and validation of an HPLC method for
determination of antidiabetic drug alogliptin benzoate in bulk and tablets. Journal of
Analytical Methods in Chemistry. 2018 Sep 24; 2018

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77

Synthesis of colloidal metal oxide using reverse micelle technique and its
application in coupling reactions for Benzoxazole formation
Ramesh N. Zade1, Pravin S. Bodakhe*, Kishor B. Raulkar2, Bhupesh M. Mude3
1. Siddharth College of Arts, Science and Commerce, Fort, Mumbai
2. Vidya Bharati Mahavidyalaya, Camp Amravati
3. Vidya Bharati Mahavidyalaya, Camp Amravati
4. Ramanarain Ruia College, Matunga, Mumbai

Abstract:
Metal oxides have profound applications in the various advanced technologies using its
catalytic, gas sensing, optoelectronic, ceramic, piezoelectric properties. Physiochemical
properties of metal oxides are tuned using various synthetic methods to synthesize particles of
various shapes, sizes and geometries. Therefore, focus of the researchers and scientists is to
prepare metal oxides nanoparticles using various synthetic routes of Chemical, physical and
biological methods. But either these synthetic routes are complicated, energy intensive or
producing nanoparticles having potent toxicity which can restrict its long-term use in the
various applications and technologies. Potential toxicity of nanoparticles is due to its
accumulation in vital organs of the body. Exposure to nanoparticles could trigger the
production of reactive oxygen species which subsequently disturb the metabolism as well as
damage proteins, enzymes, cell membranes and also DNA. Therefore, for sustainable
development of nanotechnology use of nanoparticles in consumer products, environmental
benign techniques for the synthesis of metal oxides are to be practiced. Colloidal metal oxides
particles have shown promising results in this direction, as colloids are biocompatible in its
synthetic ways as well as its application modes. Therefore, in this article, we have focused to
prepare copper oxide and nickel oxide using some synthetic methods which successfully lead
to synthesize colloidal metal oxides. Characterization of synthesized metal oxide is done for
its shape, size, morphology using techniques such as SEM, XRD, FT-IR and BET. These metal
oxides subsequently used for reactions which forms the basis of complicated reactions taking
place in plant and animal body. These reactions are mainly coupling reactions involving C-N
bond and C-O bond formation.
Key words: Metal oxide, nanotechnology, toxicity, reverse micelle, Coupling reactions,
Benzoxazole
1. Introduction:
Metal ions are fundamental elements present in plants and animals. Their substantial role in
biological systems was recognized a long time ago. They are essential for the maintenance of
life and their absence can cause growth disorders, severe malfunction, carcinogenesis or death.
They are protagonists as macro or microelements in several structural and functional roles,
participating in many biochemical reactions, and arise in several forms. They participate in
intra and inter cellular communications, in maintaining electrical charges and osmotic pressure,
in photosynthesis and electron transfer processes, in the maintenance of pairing, stacking and
the stability of nucleotide bases, and also in the regulation of DNA transcription. They
contribute to the proper functioning of nerve cells, muscle cells, the brain and the heart, the
transport of oxygen and in many other biological processes up to the point that we cannot even
imagine a life without metals.[1]
Heterocyclic moieties play a significant role in the field of drug discovery. C−N and C−O bond
formation reactions are the primary synthetic sequence for the generation of heterocyclic
molecules. The generation of C−N and C−O bonds involves the use of mostly Pd or Cu

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catalysts although other transition metal catalysts are also involved. However, in C−N and C−O
bond formation reactions, several problems were faced such as catalytic systems containing
costly ligands, lack of substrate scope, lots of waste generation, and high temperature
conditions. So it is imperative to uncover new eco-friendly synthetic strategies which provides
a short reaction time, tolerance for functional groups, and less waste production. [2, 8, 9]
Microbial organisms possess an incredible capability of fabricating extremely specialized
inorganic nanostructures. These magnificent skills of the living creatures have caught the
attention of many material scientists towards these biological systems to acquire knowledge
and recuperate the skills for the precise formulation of nanostructures. Generally, synthesis of
inorganic nanomaterials by living organism has been categorized as biologically controlled and
biologically induced synthesis. Biologically controlled synthesis is well-known to occur
naturally in a few organisms. During this type of synthesis, the organisms are able to modulate
the particle size, composition and surface area of the produced particles. Though biologically
controlled synthesis of metal oxide nanoparticles exhibits high control over the morphology
and composition of the nanoparticles. [3,4]
The basic idea of the colloidal method is to limit particle growth. As the formation of very
small crystals is thermodynamically unfavourable due to the high surface energy of such
particles, nanosynthesis must be controlled by kinetics. To achieve this, surfactant molecules
are used to limit particle growth. One method uses a non-polar solvent, leading to the formation
of reversed micelles. In the polar core of the micelle, a minute amount of water is present in
which precipitation of the nanocrystal takes place. The size of the micelles thus determines the
size of the particles. [5,6]
Benzoxazoles are an important class of heterocyclic compound that are encountered in a
number of natural products and are used in drug and agrochemical discoveries. As a
consequence, much effort has been devoted not only to construct basic skeleton of benzoxazole
molecules but also to prepare derivatives through C-O or C-N atom bond formation. Hence
development of an efficient methodology for the preparation of benzoxazole and its derivatives
has gained a lot of importance in current research. [10,11]
2. Synthesis and characterisation of MO:
Cuprous oxide (Cu2O) is produced by chemical reduction of copper sulphate salts in water-in-
oil microemulsion solution using NaBH4 as a reductant. Water-in-oil (w/o) microemulsions
also called reverse micelles are water pools stabilized by surfactants, dispersed in oil phase,
that serves as nanoreactors in which nanoparticles form and serve in controlling the size of
nanoparticles. The water pools being stabilized by the lipopeptidal surfactant act both as
nanoreactors for the process reaction and prevent particle aggregations as the surfactants get
adsorbed on the particle surfaces when the particle size approaches the water pool, resulting in
fine and uniform particle size distribution. Colloidal particles were then supported on γ-Al2O3
are then dried in oven at 800C for an hour. Characterisation of this colloidal metal oxide is done
by FTIR, SEM and XRD.

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Figure 1 : SEM micrographs at different O/W ratio.

Figure 2: XRD

Figure 3: FTIR

3. Application of MO:
CuO prepared is further used as catalyst for coupling reaction in the formation of new C-N
and C-O bonds. To illustrate this application following coupling reaction is taken where
biologically important Benzoxazole is prepared using amide and aryl diazonium salt.

Conditions of temperature and solvents are optimised at 2 hours of heating and effects
of equivalents of CuO is shown below
Sr.No Equivalents of catalyst Temperature ( oC) Yield (%)
1 1 110 68
2 2 110 85
3 3 120 85
4 4 120 82

Reaction conditions: amide(1mmol), diazonium salt (1mmol), catalyst (1mol %), solvent
( 2 mL), KI (1.5mmol), K2CO3 (3mmol) 1100C, time 2 Hrs; isolated yield
4. Result and Conclusion: Efficient, economical, base free protocol for benzoxazole
synthesis has been developed with considerable good yield, high functional group
tolerance, cost effective (due to use of diazonium salts). Moreover, Cuo NP used is prepared

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by reverse micelle which is biocompatible. Therefore, it has prospectus to use in biological

systems for diagnose metabolism and also for curative action in case of diseases and
deficiencies.
References:
1. Materials (Basel). 2021 Feb; 14(3): 549. Published online 2021 Jan 24. doi: 10.3390/ma14030549
PMCID: PMC7866148 PMID: 33498822 The Role of Metal Ions in Biology, Biochemistry and
Medicine, Michael Moustakas
2. Copper Catalyzed Synthesis of Heterocyclic Molecules via C−N and C−O Bond Formation under
Microwaves: A Mini-Review Sushovan Jena and Kaushik Chanda
3. Toxicity of metal and metal oxide nanoparticles: a review, Ayse Busra Sengul & Eylem
Asmatulu Environmental Chemistry Letters volume 18, pages1659–1683 (2020)
4. Metal oxide nanoparticles and their applications in nanotechnology Murthy S. Chavali, ·
Maria P. Nikolova Springer Nature Switzerland AG 2019
5. Colloidal synthesis of metal oxide nanocrystals and thin films Fredrik Söderlind, Linköping Studies in
Science and Technology Dissertation No. 1182.
6. Colloidal Metal Oxide Nanoparticles Synthesis, Characterization and Applications, Metal Oxides, 8 -
Colloidal metal oxides in energy technologies, Sungwook Chung 2020, Pages 183-201
7. Polymer-Metal Oxide Composite (PPy–MoO3) for Ammonia and Ethanol Gas Sensor, Surendra M.
Yenorkar, Ramesh N. Zade, Bhupesh M. Mude, Vijay M. Mayekar, Kushal M. Mude, Kishor B. Raulkar,
Ranjeet R. Mistry, A.N. Patange, Macromolecular SymposiaVolume 400, Issue 1/2100049, 22 December
2021.
8. ASeung Hwan Cho, Ji Young Kim, Jaesung Kwak and Sukbok Chang* Chem. Soc. Rev., 2011, 40,
5068–5083
9. Xiao-FengWua*, Helfried Neumann , Stephan Neumann , Matthias Beller, Tetrahedron Letters 54
(2013) 3040–3042
10. Sachin A. Sarode, Jeevan M. Bhojane, Jayashree M. Nagarkar*,Tetrahedron Letters 56 (2015) 206–210
11. Russell D. Viirre, GhotasEvindar, and Robert A. Batey*, J. Org. Chem. 2008, 73, 3452–3459

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78

Synthesis, Charecterization and Biological Evaluation of Coumarin –

Chalcone Derivatives

Disha M. Dhabarde, Punam B. Rathi, Ashish Telrandhe

[email protected]
Kamla Nehru College of Pharmacy, Butibori, Dist.-Nagpur, 441122

ABSTRACT –
In the present study, an attempt has been made to synthesize and characterize some coumarin-
chalcone derivatives and to evaluate them for their anti-microbial activity. The compounds
were prepared as per the reported procedure in literature. The physicochemical characteristic
like melting point, percentage yield, Rf value were evaluated. The synthesized compounds
were characterized by IR, NMR and Mass spectroscopy. Compounds exhibited good
antimicrobial activity when compared with standard. Further, there is a large scope for the
development of other derivatives and their pharmacological screening. Since, the synthesized
scheme is simple and now well established it would be easy to synthesize other derivatives by
incorporating various substitutions and screenings them for other pharmacological activities
like Antibacterial, antiviral, anti-tumor, anti-hyperglycemic, analgesic, anti-inflammatory,
Moreover the newly synthesized Derivatives can be subjected for QSAR analysis and docking
to emphasize the Pharmacophoric requirement and to study drug-receptor interaction and
binding for specific target.

Keywords – Chalcone derivatives, Coumarin, Anti-microbial activity

INTRODUCTION –
Coumarin nucleus is widely distributed in nature in plant kingdom and forms an important
class of oxygen heterocycle1. Over the years, coumarins have been established as well-known
naturally occurring oxygen-heterocyclic compounds isolated from various plants2. They are
the family of lactones containing benzopyrone skeletal framework that have enjoyed isolation
from plant as well as total synthesis in the laboratory. The plant extracts containing coumarin-
related heterocycles are employed as herbal remedies in traditional systems of medicine. The
synthesis of coumarin derivatives has attracted considerable attention of organic and medicinal
chemists due to its wide usage in food additives, fragrances, pharmaceuticals and
agrochemicals. Given this, coumarins have attracted intense interest in recent years because of
their diverse pharmacological properties. Hence, many researchers have reported different
biological activities of coumarins derivatives such as antibacterial, antiviral, antitumor, anti-
hyperglycemic, analgesic, anti-inflammatory activities and other pharmacological activities3-
11
. Chalcones are α,β -unsaturated ketones which constitute an important group of natural
products that serve as precursors for the synthesis of various heterocyclic compounds like
pyrimidines, imidazoles, pyrazoles, 2- pyrazoline and flavonoids12,13. Cyclization of chalcones,
leading to thiazines, pyrimidines, pyrazoline has been a developing field within the realm of
heterocyclic chemistry for the past several years because of their ready accessibility and the
broad spectrum of biological activity of the products as antimicrobial, antibacterial, antifungal,
antiprotozoal, anti-inflammatory substances14. With this background it has been thought worth
to synthesize some novel heterocyclic compounds comprising of chalcone and coumarin in a
single moiety. Also, to evaluate these new compounds for their potency as an anti-microbial
agent.

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MATERIAL AND METHOD

SYNTHESIS OF COUMARIN CHALCONE DERIVATIVES

A mixture of 3-acetyl-7- hydroxy-9H-benzopyrano- 1,4-oxaxin -2,9- dione and

substituted aromatic aldehyde was stirred for 12 hr. under refluxed at temperature 60° C to
obtain coumarin chalcone. The compounds were characterized by Infra-red, Mass and NMR
Spectroscopy. In-vitro antimicrobial activities of synthesized compound were studied by pour
plate method using ciprofloxacin as standard for antibacterial and ketoconazole as for
antifungal activity respectively. The synthesized compounds were evaluated for their anti-
bacterial activity against Staphylococcus aureus and Escheria coli and antifungal activity
against Aspergillus Niger. The activity of synthesized compounds showed that synthesized
compounds exhibited good antibacterial activity and antifungal activity.

REFERENCES

1. Cintas P. Activated metals in organic synthesis. CRC Press; 2020 Jan 29.;7-10
2. Bhat BA, Dhar KL, Puri SC, Saxena AK, Shanmugavel M, Qazi GN. Synthesis and biological evaluation of
chalcones and their derived pyrazoles as potential cytotoxic agents. Bioorganic & medicinal chemistry letters.
2005 Jun 15;15(12):3177-80.
3. Monica Kachroo, Synthesis of some new chalcone derivatives and evaluation of their anticancer activity. Int. J.
Drug Dev. and Res, (2013); 5(3):309-315.
4. Mokle SS. Sayeed MA. Kothawar and Chopde. Int. J. Chem. So 2004;2(1):96-100
5. Cheng JH, Hung CF, Yang SC, Wang JP, Won SI, Lin CN. Synthesis and eytotoxic, anti-inflammatory, and anti-
oxidant activities of 2, 5% dialkoxyfchalcones as cancer chemo preventive agents. Hioorganic & medicinal
chemistry. 2008 Ang 1:16(15):7270-6
6. Seema 1. Habib et al. Chemical and biological potential of chalcone as a source of Drug: A review. LIPPR, (2018);
11(02):104-118.
7. Rajendra Prasad Yejella et al. Synthesis, antimicrobial, and computational evaluation of novel Isobutyl chalcones
as antimicrobial agents, UMC, (2017), 1- 14.
8. Sunny Jalhan et al. Various Biological activities of coumarin and Oxadiazole derivatives. Asian J. Pharm Clin
Res, (2017); 10(7):38-43)
9. Chapter 5 Synthesis of 7-hydroxy-4-methyl coumarin over Zapo-5 and Lewis acid metal ion-exchanged Zapo-5
molecular sieves 79.90.
10. Antimicrobial Merriam-Webster online dictionary. Archived from the original on (2009).
11. Alivelu Samala et al. Synthesis, characterization of some novel coumarin derivatives and evaluation of their
Pharmacological activities. Der Pharma chemical, (2016), 8(12):19-24.
12. Kubba et al. Synthesis and characterization of new coumarin derivatives containing various moieties with
antibacterial activities. Int. J. Pharm, Pharm sci, (2015); 7(8):70-74.

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79

Synthesis and Characterization of (3-(3,5-dichloro-2-hydroxyphenyl)-5-

(pyridin-2-yl) isoxazol -4-yl)(phenyl) methanone with microwave
irradiation.
P. S. Nandurkar1, M. M Rathore2
1
Department of chemistry, Sant Gadge Baba Amravati University, Amravati
2
Department of chemistry Vidyabharati Mahavidyalaya Amravati, Amravati

Abstract:
Isoxazoles1 is an unsaturated aromatic heterocyclic compound containing three carbon one
oxygen and one nitrogen in a ring atom. Isoxazole being an azole with an oxygen atom next to
the nitrogen exhibits a broad spectrum of biological activity and also forms a part of various
biodynamic agents2. A mixture of 3-benzoyl-6,8-dichloro-2-(1-phenylprop-1-en-2-yl)-4H-
chromen-4-one (VIa) (0.01 M), hydroxylamine hydrochloride irradiated with DMSO in
microwave.
Introduction:
Isoxazole being an azole with an oxygen atom next to the nitrogen exhibits a broad spectrum
of biological activity and also forms a part of various biodynamic agents. The substituted
isoxazoles are also considered to be important structure due to their versatility toward chemical
transformations to useful synthetic intermediates such as 1,3-dicarbonyl, 1,3-iminocarbonyl;
and γ-amino alcohols.
The Chlorosubstituted isoxazoles have a wide range of pharmaceutical applications. In most of
the skin creams substituted isoxazoles are the main constituents. Various kinds of skin diseases
are cured by chlorosustituted isoxazoles. The healing process of skin specially done by the
plant extract found in nature. It is interesting to notice that structure of isoxazole is found in
plant extract used for healing process.
Methods of preparation:
A mixture of 3-benzoyl-6,8-dichloro-2-(pyridin-2-yl)-4H-chromen-4-one (VIc) (0.01 mol) and
hydroxylamine hydrochloride was irradiated in DMSO (20ml) containing 0.5ml of piperidine
for 3 min. 15 sec. in microwave. After cooling the mixture, it was decomposed into the ice and
solid product thus obtained crystallized from methanol. The gray coloured crystals of the
compound (IXc) were obtained. Yield: 70%, m.p: 1910C.
Cl OH

Cl O

O N
NH2OH.HCl Cl

N
Cl DMSO
O
O O N

MW, 800 W, time: 3 min. 15 sec., Solvent: DMSO.

Microwave: 3 min. 15 sec, Yield: 70%
Conventional: 1 hour 20 min., Yield: 69%
All the synthesized compounds have been characterized on the basis of their chemical
properties and spectral data.

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1. The UV-VIS spectrum of the compound (3-(3,5-dichloro-2-hydroxyphenyl)-5-(pyridin-

2-yl) isoxazol -4-yl)(phenyl)methanone showed λmax value 355 nm respectively
corresponding to n→ π * transition.

2. The IR spectrum (Spectrum No.54) of the compound (Xa) and compound (Xd) (Spectrum
No.57) recorded in KBr showed following main absorption bands.
Compound Frequency (cm-1) Intensity Correlation
IXd 3430 vb Ar-OH stretch
Spectrum No.51
2919 vb Ar C-H stretch
1651 s C=O stretch
1438 s C=N stretch
1169 s C-O stretch
642 w C-Cl stretch

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1. The H1 PMR spectrum of the compound (II) (Spectrum No.52) recorded in CDCl3 with
TMS as an internal standard. The observed chemical shifts can be correlated as:
Compound Chemical shifts δ Name of No.of Types of
(IXd) ppm peak protons protons
Spectrum No.52
12.71 s 1H Ar-OH
6.99 to 7.64 m 9H Ar-H
Cl OH
O

Cl

N
O
N
(3-(3,5-Dichloro-2-hydroxyphenyl)-5-(pyridin-2-yl)isoxazol-4-yl) (phenyl)methanone

Conclusion and Discussion of the Result

The short reaction time and expanded reaction range were offered by the microwave assisted
synthesis. Chlorosubstituted isoxazoles need 6-9 min. exposure to microwaves incomparable
to condensation reaction where it takes 2 hours so, it was thought interesting to save time in
eco-friendly synthesis of chlorosubstituted isoxazoles.

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References:
1. Claisen.; Ber, (1891), 24, 3900.
2. Hantzsch; Ann, (1888), 130.
3. T. Gilchrist ; Heterocyclic Chemistry, (1985), The Bath press.
4. S.Chauhan, Y.Joshi; Rasayan J. Chem, (2008), Vol.1, No.3 475-480.
5. V.Mane, D.Mahajan, P. Rajput; Journal of Medicinal Chemistry and Drug Discovery,
(2017), Issue 03, Vol.02, 590-598.
6. Claisen.; Ber, (1891), 24, 3900.
7. Hantzsch; Ann, (1888), 130.
8. T. Gilchrist ; Heterocyclic Chemistry, (1985), The Bath press.
9. S.Chauhan, Y.Joshi; Rasayan J. Chem, (2008), Vol.1, No.3 475-480.
10. V.Mane, D.Mahajan, P. Rajput; Journal of Medicinal Chemistry and Drug Discovery,
(2017), Issue 03, Vol.02, 590-598.
11. S. Raja, K. Jayaveera, S. Subramanyam, A. S. Reddy and C. Prakash; IJPSR, (2016);
Vol. 7(6): 2573-2585.
12. P. Kumar, M. Behera, M. Sambaiah, V. Kandula, N. Payili, A. Shree, and S. Yennam;
Hindawi Publishing Corporation Journal of Amino Acids, (2014) Article ID 721291, 14.
13. M. Maczynski, A. Drynda, B. Obminska-Mrukowicz, S. Ryng; Immunopharmacology
and Immunotoxicology, 37, (2015), 148-157.
14. J. Shaw, B. Chen, J. Bourgault, H. Jiang, K. Narendra, M. Jayshree, A. Frederick, M. Joe,
B. Kevin, P. Halina, E. Matthew and R.Peter; Am J Biomed Sci., (2012), 4(1), 14-25.
15. D. Kashinath, S. Nagaraju, N. Satyanarayana, B. Paplal, A. Vasu, S. Kanvahb; RSC Adv,
(2015), 5, 81768.
16. N Seelam , S. Shrivastava , S. Prasanthi, S. Gupta; Journal of Saudi Chemical Society,
(2016), 20, 411–418.
17. E. El-Sawy , A. Mandour, S. Hallouty, K. Shaker, H. Abo-Salem; Arabian Journal of
Chemistry, (2013), 6,67–78.
18. D. Patel, P. Kumari, N. Patel; Arabian Journal of Chemistry, (2017), 10, S3990–S4001.
19. B. Kendre, M. Landge, S. Bhusare; Arabian Journal of Chemistry, (2015),
doi.org/10.1016/jarabjc.
20. S. Kobayashi,T. Tanaka,Y. Soeda ,O. Almeid, A.Takashima; EBioMedicine, (2017), 20,
120–126.
21. L. Ramdani, O. Talhi, N. Taibi,L. Delort, C. Decombat, A. Silva, K. Bachari,M. vasson,
F. caldefie-chezet; Anticancer research, (2016), 36: 6399-6408. doi:10.21873/anticanres.
11237.
22. L. Bernardes , R. Rosaa, M. Moraesb L. Zimmermanna, E. Paulo, Schenkel, M. Steindel;
European Journal of Medicinal Chemistry, (2016), doi: 10.1016/j.ejmech.2017.01.029.
23. F. Manetti, A. Santucci, G. Bernardini, D. Braconi, E. Petricci; J. Med. Chem, (2017),
60, 4101-4125.
24. R. Hartmann , J. Emmerich, C. Koppen, J. Burkhart,Q. Hu, L. Siebenburger, C. Boerger,
C. Scheuer, M. Laschke, M. Menger; J. Med. Chem, (2017), 60, 5086-5098.
25. A. Merlo , G. Vilela, T. Fernandes, R. Rosa, S. Kelly, S. Kitney; Polym. Bull, (2015).
DOI 10.1007/s00289-015-1529-7.

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80

Microbial Examination of Nanoparticle of Tetra-O-acetyl-B-D-Glucosyl-

5aryl-4-Dithiobiurets.

Ashish G. Sarap, P.T. Agrawal

Department of Chemistry, Shri R. L. T. College of Science, Akola-444001 (Maharashtra) India
Email: [email protected], [email protected]

Abstract :-
This discipline focuses on the design, characterization, manufacture, and use of structures,
devices, and systems by manipulating form and size at the nanoscale scale. Recent years have
seen an increase in the branch of current study known as nanotechnology. Here we screened
for their antibacterial and antifungal activities against common pathogens like Escherichia coli,
Proteus vulgaris, Staphylococcus aureus, Pseudomonas aeruginosa, Aspergillus niger and
Penicillium. Some compounds exhibit less to good activity while some are resistant to the said
micro-organisms.

Keywords: TAG Isothiocyanate , Dithiobiurets , Nanoparticles and Antimicrobial Activities.

Introduction :- Carbohydrates are vastly diverse group of organic compounds occurring in all
known plants, animal and microbial life. The function of carbohydrate is to provide energy and
strength in plants and mammalian tissues they provide a whole variety of specialized functions
ranging from cell and organ differentiation to immune protection for new born babies.
Antimicrobial agents can be divided into many categories based on how they work and the
objectives they serve. Depending on the category of microorganisms impacted, division may
be made. Hence, substances that affect bacteria are known as bacteriostatic or bacteriocidal,
whereas substances that affect fungus are known as fungistatic or fungicidal1. The cup plate
agar diffusion method2-3 provides a simple, convenient reliable test specially applicable in
routine clinical bacteriology laboratory.
In particular, these substances have been effectively tested against a number of illnesses and
have therefore earned medical significance. Chemical substances are used for treatment of
diseases and has been known since the 1500’s. The chemical substances used for the treatment
of infectious diseases and diseases caused by the proliferation of malignant cell are called as
chemotherapeutic agents. Antibacterial agents are any chemicals that prevent microorganisms
from growing or that kill them. Despite the fact that many different compounds possess these
qualities. The phrase is often limited to chemicals that are active at concentrations adequate for
practical purposes when it is employed at sufficiently high concentrations.
Carbohydrate represents an important chemical class as many drugs and drug
intermediates4 are based on carbohydrates chemistry and many drugs such as amino glycoside
antibiotics containing carbohydrate structure.
Among all carbohydrates our interest is to synthesized nitrogen linked glucosyl
compounds due to its applications in medicinal chemistry and in many other ways5-6. Sugar
isocyanate are versatile synthetic intermediate in carbohydrate chemistry. They have attracted
considerable interest in synthetic and medicinal chemistry7-8. The glycosides have found use
as divertic agent, analgesics, antidiabetic compounds and in many other ways9. Methyl -
lactosyl can significantly reduce the formation of tumor colonies in mice10. To increase its
efficiency multivalent β-lactosyl have been synthesized in Roy’s group. Heterocyclic
derivative of sugars were found to possess anti-tumor and anti-bacterial activity. Besides these
and other pharmaceutical applications of glycosyl urecides, they also found to possess
applications in paper, textile and food industries

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EXPERIMENTAL
The research work presented deals with the study of antimicrobial activities of newly
synthesized N-glucosides against pathogenic organisms. Screening of following
compounds were carried out against the microbes like E. coli, P. vulgaris, S. aureus, P.
aeruginosa, A. niger and Penicillium.

Experiment No. 1:-1-Tetra-O-acetyl-β-D-glucosyl-5-phenyl-4-dithiobiuret. (IIIa)

To a toluene solution of tetra-O-acetyl- β -D-glucosyl isothiocyanate (0.005 M, 1.9g in 20 ml)

was added toluene solution of phenyl thiocarbamide (0.005 M, 0.76 g in 10 ml) and reaction
mixture was refluxed over boiling water bath for 3hr. Afterwards, solvent was distilled off and
sticky mass obtained as residue was triturated several times with petroleum ether afford a white
solid. It was crystallized with ethanol-water, m.p. 95°C. [Found: C, 50.30; H, 4.85; N, 7.93; S,
6.19, C22H26O9N3S2. requires; C, 50.38, H, 4.96; N, 8.01, S, 6.10%].
The product was found soluble in ethanol, acetone, chloroform and benzene
while insoluble in water and petroleum ether. It charred on heating with conc. sulphuric acid.
It was found desulphurisable when boiled with alkaline plumbite solution. It was optically
active and its specific rotation was found to be [α]D32 = -136.94° (c,0.74 in chloroform). The
purity of the product was checked by TLC, Rf value 0.69 (CCl4: EtOAc, 3:2)..

RESULTS AND DISCUSSION:-

The synthesis of N-glycosyl Dithiobiurets is a simple and reliable route. This strategy can be
successfully applied to prepare a wide range of glycosyl Dithiobiurets and their derivatives
which can be widely used for the preparation of biologically active molecules and good active
lead in Medicinal Chemistry. Thus the synthesized novel N-glycosyl Dithiobiurets exhibits
antibacterial and antifungal activities against the organisms tested. The method adopted in the
synthesis and investigation is simple, efficient and inexpensive in synthesizing
pharmacologically important molecule.
Antimicrobial Activity :- These newly synthesized thiocarbamides were screened for their
microbial activity against different pathogenic microbes for their antibacterial and antifungal
activities using well method16. The compounds were screened for antibacterial activity against
E. coli, P. vulgaris, S. aureus, P. aeruginosa, A. niger and Penicillium. in potato dextrose agar
medium. Procedure for antimicrobial screening Media used (Nutrient broth): Peptone – 10 g,
NaCl – 10 g and yeast extract 5 g, Agar 20 g in 1000 ml of distilled water. Initially, the stock
culture of bacteria were revived by inoculating in broth media and grown at 37 0C for 18 h.
The agar plates of the above media were prepared and wells were made in the plate. Each plate
was inoculated with 18 h old culture (100 µL, 104 cfu) and spread evenly on the plate. After
20 min. the wells were filled with different concentrations of samples. The control wells were

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filled with Gentamycin. All the plates were incubated at 37 OC for 24 h and the diameter of
inhibition zones were noted in mm. The activity was quantitatively assessed on the basis of
inhibition zone

Table 1 : Antimicrobial activities of 1-Tetra-O-acetyl- β -D-glucosyl-5-aryl -4

dithiobiurets (I a-g)
Compounds E.coli P.vulgaris S.aureus P.aeruginosa A.niger Penicillium
I-a +++ +++++ +++ ++++ ++++ ++
I-b +++ +++ +++ ++++ ++ +++
I-c ++ ++ +++ ++++ ++ ++++
I-d ++ ++++ +++ ++++ +++ +++
I-e ++ ++++ ++ ++++ +++ ++
I-f -- +++ ++++ ++ +++ ++++
I-g +++ +++ +++ ++++ +++ ++
N.B. : ++++ Strongly active (above 20 mm)
+++ Moderately active (15 mm to 20 mm)
++ Weakly active (8 mm – 14 mm)
-- Inactive (below 8 mm)
Bore size = 7 mm

The compounds 1-tetra-O-acetyl– β -D-glucosyl-5-aryl-4-dithiobiurets (I-7) showed

comparable activity. Compounds 1,2,3 showed strong activity against P.vulgaris and S.aureus
Penicillium. Compounds 1-7 showed moderate activity against used microorganisms.

CONCLUSION
Derivatives were synthesized and characterized for their structure elucidation. Various
chemical and spectral data supported the structures. Some of the compounds synthesized
showed promising antimicrobial activities. The newly synthesized thiocarbamides and
thiocarbamates exhibits comparable antibacterial and antifungal activities against the
organisms tested. The method adopted in this investigation is simple, efficient and inexpensive
and is useful in synthesizing pharmacologically important molecules. The method adopted in
the synthesis and investigation is simple, efficient and inexpensive in synthesizing
pharmacologically important molecules.
Acknowledgement
Authors are thankful to PDKV Akola for providing the spectral data and also to DR. V. D.
Nanoty Principal, Shri R. L. T. College of Science, Akola for providing necessary facilities

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Reference :
[1] I. Goodman.;Carbohydrate-urea-phenol-based adhesives: Transient
formation of mono- and di-D- glucosylureaAdv. Carbohydr.
Chem.,1958 13, 215-236
[2] L.T. Shih, M-C, Cheng, S-H, Wu,.Tetrahedronlett., 2002 ,44, 7921-7923
[3] Y. Mishikawa, T. Terkeda, S.Shibata, F.Fukuoka., Bull. Chem. Pharma, 1969., 17,
1910-1916
[4] A.V. Berenguel, F.O. Caballero, F.Santoya-Gonzalez, J.J.Gracia-Lopez,
J.J.Gimenez-Martinez, L.Gracia- fuentes, E.O. Salmeron., J. Eur. Chem.,
2002., 8, 812-827
[5] Gunawardana, G.P.; Kohmoto, S.; Gunasekera, S.P.; McConnel, O.J.; Koehn,
F.E. Dercitine, a new biologically active acridine alkaloid from a deep water
marine sponge, Dercitus sp. J. Am. Chem. 1988, 110, 4856–4858.
[6] Noel, S.; Cadet, S.; Gras, E.; Hureau, C. The benzazole scaffold: A SWAT to
combat Alzheimer’s disease. Chem. Soc. Rev. 2013, 42, 7747–7762.
[7] Prajapati, N.P.; Vekariya, R.H.; Borad, M.A.; Patel, H.D. Recent advances in
the synthesis of 2- substituted benzothiazoles: A review. RSC Adv. 2014, 4,
60176–60208.
[8] Kok, S.H.L.; Gambari, R.; Chui, C.H.; Yuen, M.C.W.; Lin, E.; Wong, R.S.M.;
Lau, F.Y.; Cheng, G.Y.M.; Lam, W.S.; Chan, S.H.; et al. Synthesis and anti-
cancer activity of benzothiazole containing phthalimide on human carcinoma
cell lines. Bioorg. Med. Chem. 2008, 16, 3626–3631.
[9] Heo, Y.; Song, Y.S.; Kim, B.T.; Heo, J.N. A highly regioselective synthesis of
2-aryl-6- chlorobenzothiazoles employing microwave-
promoted Suzuki–Miyaura coupling
reaction. Tetrahedron. Lett. 2006, 47, 3091–3094.
[10] Alaimo, R.J.; Pelosi, S.S.; Freedman, R. Synthesis and Antibacterial Evaluation
of 2-(Substituted Phenylureido)-4-thiocyanatobenzothiazoles. J. Pharm. Sci.
1978, 67, 281–282.

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81

Synthesized PANI/Cu-NPs / Aloe-Vera thin films Biocomposites for

ammine gas sensor stimulator.

Dr. D. B. Dupare@ *
1*@ Department of Chemistry, Shri R.G. Rathod Arts and Science College, Murtizapur, Di.Akola.

Abstract:
The synthesized polyaniline disperse Copper Nano particle (Cu-NPs) by the usage of
ultrasonicator, microwave instrumentation approach to metal salt on Aloe Vera Leaf Extract
thin film on glass subtract through chemically oxidative polymerization approach. The cupper
chloride metallic salt disperse Nano particle (Cu-NPs) Stabilized on Aloe Vera Leaf Extract.
The secondary phytochemical of Aleovera leaf plant extract polyphenols stabilizing and
capping system for the more balance the cupper metallic chlorides. We synthesized PANI-Cu
-NPs. Aloe Vera leaf extract studied their U.V., FTIR, XRD and Electronic Microscopy,
Characterization for electrochemical behavior and current -voltage characteristics. The four -
probe instrumentation system shows that the PANI-Cu-NPs modified electrode turned into
accurate conductivity and stability on glass substrate. The thin film of PANI-Cu-NPs
discovered gas sensing conduct for 10-50 ppm ammine gas sensor

Key word: Polyaniline, I-R Characterization, Thin film, PANI-Cu-NPs and Biocomposites

Introduction:
The biomaterial sciences include systematic study of engineering materials with various
application purposes. A biomaterial is an engineering material which is specific range of
properties like to evaluate the chemical, physical, mechanical and biological properties. Due to
this synthetic polymer can be merged with biomaterial including metals, ceramics, composites
and hybrid systems (1-2). The last three decay the numerous scientific efforts and publication
reported related to biomaterial. The biomaterial science is a multidisciplinary area in science
and technology included medicinal chemistry such as inert material gold, silver, and platinum,
used in the early designed bone fracture plates like acrylic methyl methacrylate polymer or
commercialized polymers such as Dacron used in the vascular implants and the heart valves
(3-4).

Polyaniline exhibits excellent characteristic such as good electrical conductivity also

environmental stability and an easy synthetic route. The synthetic biocompatible polymers
show great physicochemical properties. The polymers PANI are the most commonly
investigated IEPs because of several feature like good thermal stability, cost effectiveness of
the used aniline less expensive monomer, simple synthetic procedure and good conductivity(5).
PANI included metal salt materials have widespread application in the electronics, sensor,
smart material batteries optical devices and biomaterial (6).
In this paper we reported the synthesis PANI-Cu-NPs -Aloe Vera hybrids thin film composites
on glass substrate via simple chemical oxidative polymerization technique by using ultra
sonicator. The Cu-NPs are first synthesized through the reduction of copper chlorides salts with
Aloe -Vera leaf extracts of polyphenol. It is believed that these polyols can play a vital role as
stabilizers/capping agents in the production of the Cu-NPs nanoparticles. The Novelty of this
works is that plant extract polyols as well as flavonoids are used to stabilize the Copper particle
in formation of thin films on glass substrate that is biomaterial or composites is synthesized
and their utility for electrochemical characterizations in different application as sensor.

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2. Materials and Methods

2.1 Materials and chemical

Aniline and Ammonium oxysulphides from Sd -Fine chemical are used. Aniline distilled prior
to use. The copper (II) chloride hexahydrate (CuCl2.5H2O, >98%) was from Sd-Fine chemicals.
Aloe-Vera was bought from a local college medicinal garden. Double Distilled deionized water
(DDW) was from the analytical laboratory, Chemistry Department, was used for all aqueous
preparations.
2.2 Extraction of Aloe-Vera gel and Cu -NPs synthesis

We are collected the fresh leaves of Aloe-Vera was bought from our college medicinal
garden and remove the hard green coat from leaf by using knife, the after near about 10 g semi
liquid parts gel and 10 ml of double distilled water are mixed in 100ml beaker by using ultra
sonicator for 20 minutes. The resultant supernatant was collected, filtered and stored at 10 °C
before use. The CuNPs were prepared by adding 0.1 M to 0.01M Copper chloride solution to
the Aloe-Vera extract (supernatant at ambient temperature) in a series of 1:1,1:2,1:3,1:4,1:5
and also 5:1,4:1,3:1, 2:1 volume ratio. The mixture was hand shaken for 1 min and allowed to
stand at room temperature for 30 minutes in ultra sonicator at 40 to 70 °C to prepare the
nanoparticles. The all beaker kept in refrigerator for 24 hour at 10°C.

2.1 Synthesis of PANI-Cu NPs of Aloe-Vera films.

The optimize stoichiometric concentration of 2ml aniline and 0.05, upto 0.1 M Concentration
of Cu-NPs of Aloe-Vera extract solution added to in series of 100 ml beaker with glass substrate
to oxidized with 0.5 M ammonium peroxidisulphate under continuous stirring at room
temperature in hydrochloric acid solution and kept in ultrasonicator for 20 minutes then cool it
below -10 °C freezers for 24 hours to obtained homogenous stable thin film formation. The
thin films substrates wash with deionized water kept to dry at room temperature with drier. The
Figure shown in Fig 1.1 is that Synthesis of PANI-CuNPs of Aloe-Vera films after oxidation
the formation Biocomposites films of PANI-CuNPs and Fig 1.2 indicates the formation of thin
films of PANI--CuNPs on Glass substrate.

Fig 1.1 PANI-CuNPs of Aloe-Vera after oxidation Fig 1.2 thin films of PANI-CuNPs

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2.2 Characterization Technique of the PANI-CuNPs

The Synthesized biocomposites of PANI-CuNPs dissolved in DMSO solvent to observe U.V.-
Visible spectroscopy was recorded on UV- VIS Spectrophotometer Carry Agilent Tech at
central instrumentation cell, Department of chemistry SGBAU, Amravati for formation of
biocomposites and electronic transition is present to this biomaterial to show the
electrochemical behaviors. The same composites solvent observed FTIR spectra were recorded
on Bunker Alpha-T FT IR Spectrometer at central instrumentation cell, Department of
chemistry SGBAU, Amravati for identification of functional group present and this functional
group have vibration and rotation stretching of molecules. The XRD Spectra of dry powder
observed by powder XRD spectroscopy for find the copper metal present in this biocomposites
formation of biomaterials of Aleovera plants and metal dopant is this biocomposites. The
application point if we need morphology of biocomposites is necessary this morphology
observed by using High electron density microscopy by Department of Botany with in same
institutes. The ectro-chemical behavior, Current –Voltage characteristics, and ammine gas
sensing behavior were observed by four probe technique in Department of Physics BAMU
university.
3. Results and Discussion.
3.1 Electron microscopy of PANI-CuNPs film.
To the view of application, we observed the morphology and structure of film for gas
sensing application as well as I-V chaterization and other advance application like electronic
device like ploy membrane, biomaterial, battery electrode or capacitor we need the uniform
formation of films having their particular appearance of like tubular, granular or some Nano
size metal slat occurrence of film observed.

Fig 3.1Electron microscopy of PANI Fig 3.2Electron microscopy of PANI-

CuNPs

In Fig 3.1 observed that the Electron microscopy image of polyaniline films on glass substrate
is uniform, having porous in morphology which applicable in chemical gas sensing as well as
electrochemically behavior nature can be applicable for battery storage application as well as
chemical and biological sensor. In Fig 3.2 observed that the Electron microscopy image of
PANI-CuNPs Biocomposites of Aleovera plant which is uniform, porous dark green in
colouration, the film having some dark globular spot confirm the presence of cupper metal in
granule morphology which have active role in gas sensor application. These two images
confirm that there is formation PANI-CuNPs Biocomposites.
3.2 U.V Spectra of PANI-CuNPs film.
The U.V.-Visible spectroscopy was studied in range between 200nm to 700nm after dissolution
of PANI and PANI/Cu-NPs / Aloe-Vera thin films in DMSO solvents. The spectrum shows a
prominent peak at absorption maxima of 330 nm which has been assigned to the π→π*

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transitions of the Aloe-Vera polyols. A broad peak extending from 310-390 nm with
absorption maxima at 330 nm was seen.
The new peak formed was slightly shifted bathochromically compared to the 282 nm peak of
the Aloe-Vera extracts. This peak at 282 nm and the smaller hub at around 330 nm have been
shown to be due to the presence of the Cu-NPs nanoparticles in the Cu (II) state. It means the
reductant polyols within the matrices successfully reduced the Cu (II) ions to indicating the
formation of the Cu-NPs polyols hybrid. The PANI/ Cu-NPs / Aloe-Vera films composite films
have conducting in nature it observed band at 560 nm to 690nm due to presence of conducting
polaron of protonated ring of polyaniline. The PAni-Cu-NPs show in U.V. spectra it more
intensive peak or activated to n→π* at 684 nm to delocalization of lone pair of electron with
Cu-metal proton deprotonation polarons.
1.8 U.V.VISIBLE SPECTRA OF PANI-CuNPS and PANI
1.6
1.4A PANI-CuNPs
1.2b
PANI
1s
0.8o
0.6r
0.4p
0.2t
0
i
0 200 400 600 800 1000
Wavelength
In Fig 3.2 U.V Visible spectra of PANI-CuNPs

3.3 FTIR Spectra of PANI-CuNPs film.

FT-IR spectra of PANI-CuNPs exhibited the characteristic absorption bands of emeraldine salt
at 1518 and 1616 cm−1, which correspond to the C- C stretching vibration of benzenoid (N -B-
N) and quinonoid (N =Q= N) ring, respectively [7]. The peaks in the range of 1300–1400 cm−1
as arise from C-N stretching vibrations of the secondary aromatic amines, while the intense
absorption peak at 1193 cm−1 is associated to the B- NH -B and protonated Q =NH+-B
stretching modes of PANI chains [8].
Figure 3.3 of the hybrid PANI/Cu-NPs/Aloe-Vera bio composite materials indicated a
considerable red shift (∼15–20 cm−1) of the N-B-N and N =Q=N vibrational bands at ∼1501
and ∼1600 cm−1respectively, suggesting interaction of PANI chains with anionic
heteropolyacids. The IR spectra of the hybrid samples indicate the presence of Cu-Od (terminal
bonds) and Cu-Ob -Cu (bridge bonds between the corner-sharing CuCl octahedra; M = Cu)
stretchingvibrations at ∼972–988 and ∼879–890 cm−1 respectively [9]. The intense absorption
bands at∼1140 and ∼1080 cm−1 observed in the samples of Cu-Nps –Aleovera and PANI–Cu-
Nps -Aleovera are ascribed to the clusters, respectively. The aromatic C-N and O-H stretching
occurs at 3550cm-1 presence of aromatic Ar-OH and Ar-NH that alkolides and amino nature
of fictional groups. The C-O, C-N Stretching at 1774cm-1 and 1914cm-1. The various secondary
metabolites all confirm by the presence of their functionality stretching and bending occurs in
this FTIR spectra of PANI–Cu-Nps.

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100.0
99.4 99.5 99.6 99.7 99.8 99.9
Transmittance [%]

3895.18
3817.70
3766.28

3653.41

3521.80

3320.22
3234.56

3093.25
2991.06

2617.43
2518.97

2360.06

2233.04
2178.21
2092.71
2002.46
1944.04

1778.80
1692.82

1495.39

1306.62

1146.58

824.04

559.99
497.38
3500 3000 2500 2000 1500 1000 500
Wavenumber cm-1

C:\Users\Chemistry-IR\Documents\Bruker\OPUS_7.5.18\DATA\MEAS\Polyaniline +Aloevera + Copper Chloride.0 Polyaniline +Aloevera + Copper

21-04-2023
Chloride Instrument type and / or a

Page 1/1

Figure 3.3 of the PANI/Cu-NPs / Aloe-Vera bio composite

3.4.I -V characterization of PANI-CuNPs film.
The I-V characterization measurement of the PANI /Cu-NPs /Aloe-Vera films was
recorded by an indigenously developed computer controlled using four- probe method at room
temperature. The current–voltage (I-V) characteristics of the synthesized PANI /Cu-NPs thin
film was studied to ensure a ohmic or non ohmic behavior of the film. A linear and non lenar
relationship of the I-V characteristics shown in Fig.3.4. reveals that the PANI /Cu-NPs
composites film has at 300k ,305 k and 310k of different temperature for applied the voltage
at 0.5 to 10 voltages applied then for 300k it observed that initial from 0.5 ev potential to 3.5 it
shown liner ohmic charter but to increase in voltage from 3.5 to 7ev voltage it shown bent or
deflected in nature a then from 7 ev it stabilized and steady in nature appearance. The same
film at 305k near about same behavior but 310k it disappears the linearity its non linear in
nature. The average conductivity at 300k to applied the potential it shown in between 2.6x10-
5
s/cm to 70. 6x10-5s/cm.
I-V Charaterization at diifierent Temperature
100 300k
90
80
70 305k
60
50
310k
40
30
20
10
0
0 2 4 6 8 10 12

3.5. Ammine gas for 10 -50ppm sensitivity PANI-CuNPs film.

The synthesized having good morphology film of PANI-CuNPs film observed ammine gas
sensing conduction for low 10 ppm to change 50ppm hinger level. the films expose to 10
minutes for passing the ammine gas and observe the change in resistivity recorded by using
computer control four probe system and to observer the recovery period the again expose to
higher 50ppm ppm level. Due to present of Cu NPs biomaterial, it has recover and response
are good because the film have conducive as well as uniform and porous in condition it

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applicable to gas sensing behavior it can again expose to Carbon monoxide gas but it dangerous
to handle at our laboratory to take care safety purpose. Fig 3.5 observed that the ammonia gas
exopse for 10ppm level. 10 ppm ammonia gas exposed to gas sensor chamber for 5 minute
(300 second) and observed the change in resistivity of film, which is continuously increasing
but the for recovery of this gas the chamber is open and observered the change bin resistivity
it required 6.5 minutes (690 second) the recovery time is more as compared to response time
by using four prob method.

Conclusion:
This paper deal with study of PANI /Cu-NPs / films biomaterial composite. The
characterization techniques such as U.V, FTIR and EM analysis confirmed that Copper
chloride metal salt with Aleo Vera were successfully dispersed into the PANI matrix. Effect of
different concentration of cupper with Aleo vera on structural, electrical and properties of
PANI /Cu-NPs Biocomposites were studied. FTIR analysis showed good intermolecular
interaction between PANI Cu-NPs / Aloe-Vera matrix. The surface morphology of PANI /Cu-
NPs examined using EM analysis which is applicable for 10 ppm ammonia gas sensing
behavior. The I-V Chaterization at highest conductivity with a value of 2.6x10-5Scm−1,
observed electrochemical behavior which is linear and nonlinear behavior.
References
1. George Soupions, Pantelitsa Georgiou and Loukas Zoumpoulakis “Polymer Composite
Materials Fiber-Reinforced for the Reinforcement/Repair of Concrete
Structures”Polymers 2020, 12(9), 2058 pp 1-14;
https://doi.org/10.3390/polym12092058.
2. L S faeq “Thermal and Mechanical Properties of Polymer/Nickel Composites” 3rd
International Conference on Sustainable Engineering Techniques (ICSET 2020) IOP
Conf. Series: Materials Science and Engineering 881 (2020) 012092 IOP Publishing pp
1-8 doi:10.1088/1757-899X/881/1/012092.
3. Ewelina Mackiewicz, Tomasz Wejrzanowski , Bogusława Adamczyk-Cie´slak and
Graeme J. Oliver “Polymer–Nickel Composite Filaments for 3D Printing of Open
Porous Materials”Materials 2022, 15(4), 1360 pp 1-15;
https://doi.org/10.3390/ma15041360.
4. Hind Ahmed, Bahaa H. Rabee, Hussein Hakim, Ahmed Hashim and Saba R. Salman
“Preparation and Study of Optical Properties of (Polymer-Nickel Nitrate)
Composite”Advances in Physics Theories and Applications ,ISSN 2225-0638,Vol.20,
2013 pp 152-157.

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5. R. K. Goyal, R. Sulakhe“Study on poly(vinylidene fluoride)/nickel composites with

low percolation”Adv. Mater. Lett. 2015, 6(4), 309-317DOI: 10.5185/amlett.2015.5627.
6. M. Lahelin a, I. Aaltio b, O. Heczko b, O. Söderberg b, Y. Ge b, B. Löfgren a, S.-P.
Hannula b, J. Seppälä a “DMA testing of Ni–Mn–Ga/polymer composites”omposites
Part A: Applied Science and Manufacturing publishes Volume 40, Issue 2, February
2009, Pages 125-129 https://doi.org/10.1016/j.compositesa.2008.10.011.
7. Hafiza Vaneeza Hussain,MateebAhmad, Muhammad Tamoor Ansar , Ghulam M.
Mustafa , Saira Ishaq , Shahzad Naseem , Ghulam Murtaza , Farah Kanwal and Shahid
Atiq “Polymer based nickel ferrite as dielectric composite for energy storage
applications”Synthetic Metals, Volume 268, October 2020, 116507
https://doi.org/10.1016/j.synthmet.2020.
8. Mohd Mohsin Nizam Ansari, Shakeel Khan and Naseem Ahmad “A comprehensive
investigation of structural, thermal and electrical properties of T0.35Zn0.55Cu0.1Fe2O4 (T
= Mn, Ni) nano ferrites”Physica B: Condensed Matter,Volume 566, 1 August 2019,
Pages 86-95.https://doi.org/10.1016/j.physb.2019.05.003.
9. A. Suhasiniaand K. P. Vinod Kumar “Solubility and swelling studies of polymer
hybrid-nickel oxide nanocomposite”J. Indian Chem. Soc.,Vol. 96, January 2019, pp. 7-
8
10. Wangping Wu, Dingkai Xie, Jiaqi Huang,Qinqin Wang and Junjun Huang “Adhesion
enhancement for nickel layer deposited on carbon fiber reinforced polymer (CFRP)
composites by pretreatment processes for lightning
strike”https://doi.org/10.1080/00218464.2022.2082870.

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82

Structural and Ion-Exchange Properties of Copolymer Derived from 4, 4’-

Biphenol - Benzidine - Formaldehyde

P. P. Kalbende1*, M. S. Dhore2, S. S. Butoliya3

1*
Department of Chemistry, Jagadamba Mahavidyalaya, Achalpur-444806, Dist.-Amravati, (MS) India.
E-mail:- [email protected]
2
Department of Chemistry, G. H. Raisoni University, Amravati-444701, (MS) India
3
Department of Chemistry, Shri Ramdeobaba College of Engineering & Management, Nagpur-440 013, (MS)
India

Abstract
A new copolymer resin has been synthesized by condensation polymerization reaction
of 4,4’-Biphenol and Benzidine with Formaldehyde in presence of 2M hydrochloric acid as
catalyst. Composition of resin was determined on the basis of their elemental analysis and
further characterized by UV-Visible, Infra-red and Nuclear magnetic resonance spectroscopy
to confine the most probable structure of synthesized copolymer.
Newly synthesized copolymer was proved to be a selective chelating ion-exchanger for
certain metal ions and were studied for Fe3+, Cu2+, Ni2+, Co2+, Zn2+, Cd2+, Hg2+ and Pb2+ ions
using their metal nitrate solutions. A batch equilibrium method was employed to study the
selectivity of metal ions uptake involving the measurements of the distribution of a given metal
ion between the copolymer sample and a solution containing the metal ion. The study was
carried out over a wide pH range, shaking time and in media of different electrolytes at different
ionic strengths. Distribution ratios of metal ions were found to be increased by rising pH of the
solutions. Hence, it can be used to recover certain metal ions from waste water, for the purpose
of purification of water and removal of iron from boiler water.

Key words Copolymers, Ion-exchangers, Distribution ratio, Metal ion uptake, Distribution
ratio.

1. Introduction
Ion exchangers are often employed in chromatographic separation of cation mixture by
using complexing agents to increase the efficiency of separation. Process of ion exchange
between liquid and solid phase in which the complex equilibria are involved, includes
conventional cation exchanger resin containing sulphonate or carboxylate group, is used which
do not form the complexes with the cations present and the complexing agent is present as a
solute in liquid phase. Chelating resins having a functional group as a complexing agent fixed
to the matrix is used when either the liquid phase may or may not contain a complexing agent
as a solute, permits the solution of some difficult analytical problems like separation of similar
ions and the enrichment of trace elements in a simpler and far less tedious manner than the
methods applied so far. This challenge has encouraged the numerous attempts by analytical
chemists for synthesizing new chelating ion exchanger with definite selectivity towards certain
ions or group of ions.
The introduction of ion exchange terpolymer resins offered new possibilities for the
separation of species, and the application of various complexing agents has lead to much
extremely effective separation procedure. In an analytical as well as synthetic inorganic
chemistry, there exist a need for good chelating polymers that combines the ease of operation
of conventional ion exchangers and selectivity of organic analytical reagents. The selectivity
of most of the reagents for metals remains predominantly in their ability to form the complexes

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with certain cations. Attempts have been made in the early fifties of the twentieth century to
synthesize organic polymers which contain complex forming groups as exchanging functions.
The selective behavior of these resins is based on the different stabilities of the metal complexes
on the resin at appropriate pH values. The optimum conditions for maximum efficiency for
given separation can be established by variation of the pH with time interval.
Selective removal, separation and recovery of metals from industrial effluents is a
problem of environmental and economic concern, in consideration of the more stringent limits
enforced for pollutants control and raising costs of raw materials. Ion exchange as a
conservative technology allows for selective removal and recovery of heavy metals. Chelating
ion exchangers are suitable for this purpose in presence of one or more donor atoms as Lewis
bases which are able to coordinate most of the toxic polyvalent metal ions acting as Lewis
acids. Accordingly, the selectivity scale of different chelating exchangers toward metals
reflects the stability of the corresponding metal-ligand complexes in the liquid phase.
Toxic and heavy metal ions, traces of organic and inorganic contaminants, radioactive
nuclides are widely occurred in aquatic systems as a result of rapid globalization and
industrialization. Sorption processes such as electrodeposition, coprecipitation, solid liquid
extraction, activated charcoal etc. have been used for the removal of these contaminants. In
recent years, terpolymer ion-exchange resins have been developed due to their wide application
in waste water treatment and pollution control, hydrometallurgy, antibiotic purification and
separation of radioisotopes, identification of specific metal ion and metal recovery [1-4]. A
batch equilibrium method has gained a rapid attention of researchers because of its acute degree
of selectivity, variety of sorbent phases and enhanced hydrophilicity [5]. Some publications on
the synthesis, characterization and applications of chelating ion exchange resins from other
researchers [6-18] and from our research group on some terpolymer resins have been reported
[19-23].
Lutfor et al [6] have prepared a chelating ion-exchange resin containing amidoxime
functional group, was characterized by FT-IR spectra, TG and DSC analysis and the chelating
behavior was studied with Cu2+, Zn2+, Ni2+, Cd2+ and Pb2+ metal ions. Removal of transition
metals from dilute aqueous solution by carboxylic acid group containing absorbent polymers
has been studied by Liu Z S and Rempel G L [12].
Extensive literature is available to interpret the experimental results in the light of
practical applicability of various terpolymer resins. Ion exchange properties of terpolymer
resins derived from o-nitrophenol and thiourea with paraformaldehyde by A. Burkanudeen et
al [7]; and salicylic acid, melamine and formaldehyde by Gurnule et al [8] have been studied.
The terpolymer derived from 2,4-dichlorophenyl acrylate/8-quinolinyl methacrylate and
acrylic terpolymer derived from 8-quinolinyl methacrylate were found to be a good cation
exchanger as it has pendant ester-bound quinolinyl group [9, 10]. Terpolymers derived from
salicylic acid and formaldehyde with resorcinol [11], 2,4-dihydroxybenzoic acid-urea-
formaldehyde [13], salicylic acid-biuret–formaldehyde [14], 8-hydroxyquinoline, guanidine
and formaldehyde [15], anthranilic acid-resorcinol-formaldehyde [16], 4-
hydroxyacetophenone -biuret-formaldehyde [17] have been proved to be good ion exchangers.
A detailed study on preparation and characterization of a new class of polymeric ligand
exchangers for selective removal of trace contaminants from water has been reported by Henry
and coworkers [18].
For chelating resin, however, reported in the publications with respect to the desired
requirement is seldom presented. But only occasionally, it proved to be valuable for separation
on laboratory scale. In this context, as reported earlier [19-23], the present study has been
carried out on synthesis, characterization, selectivity and ion exchange capacity of copolymer
resin (4,4’-BP-BD-F) derived from 4,4’-Biphenol (4,4’-BP) and Benzidine (BD) with
Formaldehyde (F).

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2. Experimental
2.1 Materials
The chemicals, 4,4’-biphenol and benzidine from Acros Chemicals, Belgium and
formaldehyde (37%) from S. D. Fine Chemicals, India of analytical grade purity were
purchased. All the solvents like N, N-dimethylformamide, dimethyl sulphoxide,
tetrahydrofuran, acetone, diethyl ether were used after double distillation. All the metal nitrates,
indicators, electrolytes and disodium salt of ethylenediaminetetraacetic acid of analytical grade
purity were used, procured from Merck, India.
2.2 Synthesis of 4,4’-biphenol-benzidine-formaldehyde (4,4’-BP-BD-F) copolymer resin
A new copolymer was prepared by condensation polymerization reaction as shown in
Figure 1, by using the molar proportion 3:2:8 of reacting monomers i.e. 4,4’-biphenol (5.58
gm, 0.3 mol) and Benzidine (4.00 gm, 0.2 mol) with formaldehyde (30 ml, 0.8 mol) in presence
of 2M HCl as a catalyst at temperature 130 oC in an oil bath for about 3 h of continuous heating
with occasional shaking. Dark reddish brown colored solid product was immediately removed,
filtered and purified by dissolving in 1:1 (v/v) conc. NaOH/water with constant stirring and
then filtered. The resulting polymer sample was washed several times with boiling water and
dried in a desiccator at room temperature. Further dried polymeric sample extracted with
diethyl ether to remove the excess 4,4’-biphenol-formaldehyde copolymer which might be
present along with the 4,4’-BP-BD-F copolymer. Finally, the terpolymer was passed through
300-mesh size sieves and kept in a vacuum over silica gel.
OH

2M HCl
3n + 2n NH2 NH2 + 8n CH2O
1300C, 3hrs
Formaldehyde

Benzidine

OH
4,4'-biphenol

OH OH OH

NH NH *

NH *
NH

OH OH
OH
n
Fig. 1. Chemical reaction of 4,4’-BP-BD-F copolymer.

2.3 Characterization of copolymer by spectral studies

Terpolymer was subjected to elemental analysis for carbon, hydrogen and nitrogen on
Perkin Elmer 2400 Elemental Analyser. UV-Visible spectra were recorded by preparing
solution in dimethylsulphoxide on Shimadzu, UV-Visible double beam spectrophotometer in
the range of 200-850 nm. Infrared spectra was recorded using KBr pellet in nujol mull on
Perkin-Elmer-spectrum RX-I spectrophotometer in the range of 4000-500 cm-1. 1H-NMR
studies were performed in solvent, dimethylsulphoxide, on Bruker Advance-II 400 MHz proton

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NMR spectrophotometer. All the analytical and spectral studies were carried out at
Sophisticated Analytical Instrumentation Facility (SAIF) Punjab University, Chandigarh.
2.4 Study of ion-exchange properties
As the batch equilibrium method has gained a rapid attention of researchers because of
its acute degree of selectivity, variety of sorbent phases and enhanced hydrophilicity [5, 19-23]
and hence has been applied to study the ion-exchange properties in the present investigation.
2.4.1. Determination of metal ions uptake in presence of electrolyte at different concentrations
The copolymer sample (25 mg) was suspended in an electrolyte solution (25 ml) of
known concentration and the pH of the suspension was adjusted to required value by using
either 0.1 M HNO3 or/and 0.1 M NaOH, and was stirred for 24 h at 30 ºC. To this suspension
2 ml of 0.1M solution of the metal ion was added and adjusted to required pH value, and then
again stirred at temperature 30 oC for 24 h and filtered. The filtrate and washing of the solid
product were combined together for the determination of metal ion content by titration against
standardized ethylenediaminetetraacetic acid solution. The amount of metal ion taken up by
copolymer was calculated from the difference between a blank reading and experimental
reading. The experiment was repeated in the presence of several electrolytes at various ionic
strength.
2.4.2 Evaluation of rate of metal ions uptake
Series of experiments, of the type described above, were carried out in order to estimate
the time required to reach the state of equilibrium under the given experimental conditions, in
which the metal ion taken up by the chelating resins was estimated from time to time, at
temperature 30 oC, in presence of 25 ml of 1 M NaNO3 solution. It was assumed that, under
the given conditions, the state of equilibrium was established within 24 h. The rate of metal
ions uptake is expressed as percentage of the amount of metal ions taken up after a certain time
related to that at the state of equilibrium, using following relationship.
𝐴𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝑚𝑒𝑡𝑎𝑙 𝑖𝑜𝑛 𝑎𝑏𝑠𝑜𝑟𝑏𝑒𝑑
𝑀𝑒𝑡𝑎𝑙 𝑖𝑜𝑛 𝑢𝑝𝑡𝑎𝑘𝑒 = × 100
𝐴𝑚𝑜𝑢𝑛𝑡 𝑜𝑓 𝑚𝑒𝑡𝑎𝑙 𝑖𝑜𝑛 𝑎𝑡 𝑒𝑞𝑢𝑖𝑙𝑖𝑏𝑟𝑖𝑢𝑚

2.4.3 Distribution of metal ions at different pH

The distribution of each one of the eight metal ions i.e. Fe3+, Cu2+, Hg+2, Ni2+, Co2+,
Zn2+, Cd2+ and Pb2+ between the polymer phase and the aqueous phase was determined at
temperature 30 oC in presence of 1 M NaNO3 solution. The experiments were carried out as
described above at different pH values and the distribution ratio (D), as defined, was calculated
using following relationship.
Weight (mg ) of metal ion taken up by 1 gm of terpolymer
D
Weight (mg ) of metal ions present in 1ml of solution
3. Results and discussion
The resin sample was dark reddish brown in color and the solubility was tested in
commonly used solvents and found to be soluble in DMF, DMSO, THF and pyridine without
any precipitation and degradation. It was characterized by elemental and various spectral
analyses and further studied their ion exchange behavior.

3.1. Characterization of terpolymer resin

3.1.1. Elemental analysis
Elemental analysis was carried out for Carbon, Hydrogen and Nitrogen contents which
agrees well with theoretically calculated values, as compared in Table 1 and derived its
empirical formula (C68H54N4O) and empirical formula weight (1054).

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Table 1. Elemental analysis data of terpolymer.

Elemental Analysis (%)
Monomer Empirical
Terpolymer empirical formula C H N
formula weight
Cal. Exp. Cal. Exp. Cal. Exp.
4,4’-BP-BD-F
C68H54N4O6 1022 77.41 76.62 5.12 5.02 5.31 5.24

3.1.2. UV-Visible spectra

Pure DMSO solvent was used to record the UV-visible spectrum of terpolymer sample
in the wavelength region 200-850 nm and shown in Figure 2. The spectrum displayed two
absorption maxima in the region 255 and 298 nm. The former and more intense band may be
due to n→π* transitions [24, 25] observed at 255 nm, indicates the presence of oxybiphenyl
moiety. Hypsochromic shift observed in biphenyl may be due to introduction of -O- group
between two phenyl groups which destroys the conjugation. Later and less intense band
observed at 298 nm is accounted for π →π* transition which may be due to biphenol group.

Fig. 2. UV-Visible spectrum of terpolymer.

3.1.3. FT-IR spectra
The FT-IR spectrum of terpolymer is represented in Figure 3 which shows a broad band,
appeared at 3400.8 cm-1, may be assigned to stretching vibration of the phenolic -OH groups,
exhibiting intermolecular hydrogen bonding [26, 27]. Presence of weak peak at 2925.5 cm -1
describes the -NH- stretch in benzidine moiety which is present in terpolymeric chain [27].
Sharp and weak peak obtained at 2856.1 cm-1 indicates the presence of stretching vibrations of
methylene -CH2- group in the terpolymer chain [26]. Stretching vibration of >C=C< in
aromatics is represented by a medium band displayed at 1662.8 cm-1. C-O stretching in phenol
is represented at 1250.5 cm-1. Bending, wagging and rocking vibrations can be accounted by
the presence of bands appeared at 1460 cm-1,1345-1350 cm-1, 750-770 cm-1 respectively, may
be due to presence of methylene bridges (-CH2-) in the polymeric chain [28, 29]. The weak
band appearing at 1010.5 and 837.7 cm-1 indicates the presence of tetrasubstitution in aromatic
ring [27].

Fig. 3. FT-IR spectrum of terpolymer

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3.1.4. 1H- NMR spectra

Solvent DMSO-d6 was used to record 1H-NMR spectrum and is represented in Figure 4;
which reveals different pattern of peaks, since each of it possesses set of proton having different
proton environment.
A significant downfield in chemical shift of proton of phenolic -OH group, observed at δ
7.9 ppm is due to intermediate proton exchange reaction of phenolic -OH group [28]. Weak
singlet is observed at δ 7.4 ppm is due to aromatic protons. Doublet signal observed at δ 6.8
ppm may be attributed to methylene protons of Ar-CH2-NH- moiety. In 4,4’-oxydianiline
moiety, the doublet observed in the region δ 7.2 and δ 7.3 ppm is attributed to protons ortho
to –NH and protons ortho to -O- respectively. A broad singlet observed at δ 6.7 ppm may be
assigned to amino proton of -Ar-NH-CH2 moiety and methylenic protons of Ar-CH2-Ar moiety
may be recognized as signal appearing at δ 3.4 ppm [27, 28].

Figure 4. 1H-NMR spectrum of terpolymer

On the basis of the nature and reactive position of the monomers, elemental analysis,
UV-visible [24, 25], FT-IR [26, 27] and 1H-NMR [27, 28] spectra, the most probable structure
has been proposed as shown in Figure 1.
3.2 Study of ion exchange properties
To suggest the utility of newly synthesized terpolymer resins as ion exchanger for
various applications, the study of influence of various electrolytes on the selectivity of metal
ions, the rate of metal ion uptake and the distribution ratio of metal ions between the terpolymer
and solution containing the metal ions, have been carried out. In a view to ascertain the
selectivity of terpolymer resin, as ion exchanger, studies for the selected metal ions using batch
equilibrium technique developed by Gregor et al and De Geiso [31] et al, have been carried out.
The results of batch equilibrium study are presented in Figures 5 to 10 in which aqueous
solution of metal nitrate of Fe3+, Cu2+, Hg+2, Ni2+, Co2+, Zn2+, Cd2+ and Pb2+; and electrolytes
NaNO3, NaCl, NaClO4 and Na2SO4 were used. The ion exchange study was carried out using
three experimental variables namely; a) Electrolytes at different ionic strength b) Shaking time
and c) pH of the aqueous medium by keeping the two parameters constant. Only one parameter
was varied at a time to evaluate its effect on metal ion uptake capacity of the copolymer [30,
31].
3.2.1 Effect of Electrolytes and their concentrations on metal ions uptake capacity.
The influence of ClO 4- , NO3- , Cl- and SO42- at various concentrations on the
equilibrium of metal-resin interaction has been studied and the results are expressed graphically
shown in Figures 5 to 8. It shows that the amount of metal ions taken up by a given amount of
copolymer depends on the nature and concentration of electrolyte present in solution. In
presence of ClO4-, Cl- and NO3- ions, the uptake of Fe+3, Cu+2, Hg+2, Zn+2, Cd+2, Co+2, Ni+2 and

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Pb+2 ions increases with rise in concentration of the electrolytes. Further, it was observed that
in presence of SO4-2 ions, the amount of above metal ions taken up by the copolymers decreases
with increasing concentration of electrolytes.
Moreover, higher increase in uptake of Fe+3, Hg+2 and Pb+2 ions with increasing
concentration of ClO4-, Cl- and NO3- ions has been observed as compared to Cu+2, Zn+2, Cd+2,
Co+2 and Ni+2 ions, but it decreases in presence of SO4-2 ions. It was considered that the anions
like ClO4-, Cl- and NO3- form weak association with the above metal ions while SO4-2 ions may
form stronger complexes. Thus, this association may affect the equilibrium which may be
explained on the basis of stability constants of complexes with these metal ions and the results
agrees well as reported earlier [19, 20].

Fig. 5. Evaluation of the influence of different electrolytes on uptake of several metal

ionsa by terpolymer resin at different concentrations of NaNO3 electrolyte solution.
a
[M(NO3)2]=0.1 mol/l; volume of metal ion solution = 2 ml; volume of electrolyte solution =
25 ml; weight of resin = 25 mg; time = 24 h; at room temperature

Fig. 6. Evaluation of the influence of different electrolytes on uptake of several metal

ionsa by terpolymer resin at different concentrations of NaCl electrolyte solution.

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Fig. 7. Evaluation of the influence of different electrolytes on uptake of several metal ionsa by
terpolymer resin at different concentrations of NaClO4 electrolyte solution.

Fig. 8. Evaluation of the influence of different electrolytes on uptake of several metal ionsa by
terpolymer resin at different concentrations of Na2SO4 electrolyte solution.

3.2.2 Rate of uptake for metal ions as a function of time

The rate of metal adsorption refers to the change in concentration of metal ions in
aqueous solution which is in contact with given polymer and was determined to find out the
shortest period of time for which equilibrium could be carried while operating as close to
equilibrium conditions as possible. The results are expressed graphically shown in Figure 9,
show that the time taken for the uptake of different metal ions at a given stage depends on the
nature of metal ion under given conditions. It is found that Fe3+ ions required about 3 h for the
establishment of the equilibrium, whereas Cu2+, Ni2+, Co2+ and Zn2+ required 5 h and Hg+2,
Cd2+ and Pb2+ ions required about 7 h. Thus, the rate of metal ions uptake follows the order as
Fe+3 > Ni+2 > Pb+2 > Cu+2 ≈ Co+2 > Cd+2 > Zn+2 > >Hg+2 which agrees well with the results reported
in literature [19, 20].

Fig. 9. Comparison of the rate of metal ionsa uptakeb by terpolymer, a [M(NO3)2] = 0.1 mol/l;
volume : 2 ml; NaNO3 = 1.0 mol/L; volume: 25 ml, room temperature.
b Metal ion uptake = (Amount of metal ion absorbed x 100) / amount of metal ion absorbed at

equilibrium.
3.2.3 Distribution ratio of metal ions at different pH
The effect of pH on the amount of metal ions distributed between two phases can be
explained on the basis of results shown in Figure 10. The distribution ratio as a function of pH
indicates that the relative amount of metal ion taken up by the terpolymer increases with rising
pH of the medium, however, the magnitude of increase is different for each metal ion. The
study was carried out only up to pH 6.5 in order to prevent hydrolysis of the metal ions at
higher pH. In case of Fe3+ the highest working pH is 2.5. The terpolymer resin taken up Fe3+
ion more selectively than any other metal ions under study. The order of distribution ratio of

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metal ions measured in pH range 2.5 - 6.5 is found to be as Fe+3 > Cd+2 > Co+2 > Zn+2 > Cu+2 >
Hg+2 > Pb+2 ≈ Ni+2.
Thus, the results of this type of study are helpful in selecting the optimum pH for a
selective uptake of a particular metal cation from a mixture of different metal ions [24]. For
example, the result suggest the optimum pH 6.0, for the separation of Co+2 and Ni+2 with
distribution ratio ‘D’ at 3000.0 and 568.90 respectively, using the present terpolymer resin as
ion-exchanger. Similarly, for the separation of Cu+2 and Fe+3 the optimum pH is 3 at which the
distribution ratio for Cu+2 is 1466.6 and that for Fe+3 is 3900.0. The lowering in the distribution
of Fe+3 was found to be small and hence, efficient separation could be achieved.
In order to assess the utility of present terpolymer resin for the separation of Fe3+ ions
from other metal ions, the following combinations of metal solutions were prepared: (1) Fe3+
and Cu2+ (2) Fe3+ and Ni2+ (3) Fe3+ and Co2+ (4) Fe3+ and Zn2+ (5) Fe3+ and Cd2+ (6) Fe3+ and
Pb2+ etc. The solution for separation studies were prepared by mixing 1ml of 0.1M solutions
of Fe3+ with 1ml of 0.1M solution of Cu2+, Ni2+, Co2+, Zn2+, Cd2+ Hg+2 and Pb2+. Selective
uptake of the metal ions was studied by adjusting the optimum at pH 3. Distribution ratios of
Fe3+ at pH 3 in the mixture with metal ions Cu2+, Ni2+, Co2+, Zn2+, Cd2+ and Pb2+ were found
to be 1466.6, 568.90, 3000.0, 2537.0, 3700.0 and 870.70 respectively i.e. slightly lower than
3900.0 found when Fe3+ alone was studied. The lowering in the distribution ratios of Fe3+ was
found to be small and hence efficient separation could be achieved which shows the importance
of newly synthesized copolymer resins as an effective ion exchanger and can be explored for
the separation of other metal ions.

Fig. 10. Distribution ratio, Da, of various metal ionsb as function of the pH by terpolymer
aD = weight (in mg) of metal ions taken up by 1g of terpolymer/weight (in mg) of metal ions

present in 1ml of solution.

b[M(NO ) ] = 0.1 mol/l; volume : 2 ml; NaNO = 1.0 mol/l; volume: 25 ml, time 24 h
3 2 3
(equilibrium state) at room temperature.

4. Conclusion
A synthesis of new terpolymer 4,4’-BP-BD-F, based on the condensation reaction of
4,4’-biphenol and benzidine with formaldehyde in presence of acid catalyst was carried out
and structure has been confirmed by elemental and spectral studies. It was proved to be
selective chelating ion-exchanger for Fe3+, Cu2+ Ni2+, Co2+, Zn2+, Cd2+, Pb2+ and Hg+2 metal
ions and has been successfully used for selective separation of ferric ions from other metal ions
having combinations like a) ferric and cupric ion b) ferric and mercuric ion c) ferric and zinc
ion and d) ferric and lead ion etc. This polymer showed higher selectivity for Fe3+ at pH 2.5 as
compared to other metal ions.
To explore the use of this chelating resin for the separation of other metal ions in
analytical chemistry appears to be advantageous when the separation problem can not be solved

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by simpler means or when time can be saved. The area of application especially includes the
concentration and the separation of chemically very similar ions. The use of this eco-friendly
ion exchanger resin may effectively be explored in hydrometallurgy, antibiotics, purification
and separation of radioisotope, in water treatment and pollution control, for the removal of
hazardous and toxic metal ions like other ion exchangers.
Acknowledgement
The authors wish to express their sincere thanks to Principal, Jagadamba
Mahavidyalaya, Achalpur for constant encouragement and support.

References
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83

Navigating the Therapeutic Landscape: Discovery and Pharmacophore

Modeling of BRD4 Inhibitors for Prostate Cancer Precision Treatment

Mr. P S. Nawale1, Ms. A D. Mehakare2, Dr. J R. Bansod3, Prof. Capt. M M. Rathore4.

1. Department of chemistry, Vidya Bharati Mahavidyalay camp, Amravati
2. Department of chemistry, Vidya Bharati Mahavidyalay camp, Amravati
3. Department of chemistry, Vidya Bharati Mahavidyalay camp, Amravati
4 Department of chemistry, Vidya Bharati Mahavidyalay camp, Amravati

Abstract:-
This study represents a ground breaking exploration in the realm of prostate cancer therapeutics
by focusing on the identification and optimization of inhibitors targeting Bromodomain-
containing Protein 4 (BRD4). The research meticulously navigates the intricacies of BRD4
inhibition with a specific emphasis on tailoring these inhibitors for the treatment of prostate
cancer. Employing a multifaceted approach, we delve into the process of identifying novel
BRD4 inhibitors, breaking new ground in the field of cancer research. The study's core
methodology involves not only the identification and optimization of BRD4 inhibitors but also
their customization for precision treatment of prostate cancer. Pharmacophore modelling
emerges as a pivotal tool in this endeavour, providing insights into the structural features
crucial for effective inhibition. The interplay between innovative drug design and
computational modelling techniques promises a refined understanding of the intricate
molecular interactions between BRD4 inhibitors and their target. Through a comprehensive
analysis, this research aims to contribute to the development of advanced therapeutic strategies
for prostate cancer. The outcomes hold the potential to revolutionize precision medicine in the
context of cancer treatment, offering new avenues for tailored therapeutic interventions. This
study not only breaks new ground in the identification and optimization of BRD4 inhibitors
but also paves the way for future advancements in prostate cancer therapeutics through the
integration of pharmacophore modelling.
Keywords: Pharmacophore modelling, BRD4, Drug Discovery
1. Introduction:
Bromodomain-containing protein 4 (BRD4) is a crucial regulator involved in transcriptional
and epigenetic processes, playing a pivotal role in various cellular functions and diseases. As
a member of the BET (bromodomain and extra-terminal domain) family, BRD4 is known for
its influence on gene expression and chromatin remodeling [1]. It has been extensively studied
in the context of cancer, where it acts as a transcriptional and epigenetic regulator impacting
cancer progression and inflammatory diseases [2][1]. Additionally, BRD4 has been associated
with angiogenesis and is recognized as a potential therapeutic target for diseases involving
abnormal cell growth [3] [4]. The interplay between BRD4 and the MYC proto-oncogene
further highlights its significance in both normal development and disease pathogenesis [5].
BRD-4 inhibitors have emerged as promising therapeutic agents in various medical contexts.
These inhibitors target the bromodomain-containing protein 4 (BRD-4) and are designed to
modulate its activity. Treatment with BRD-4 inhibitors has been shown to reduce the
expression of super-enhancer-associated oncogenes like MYC, E2F1, and BCL6, highlighting
their potential in cancer therapy [6]. Moreover, BRD-4 inhibition has been associated with
impaired DNA mismatch repair, induction of mismatch repair mutation signatures, and the
creation of therapeutic vulnerabilities, particularly in immune-related responses [7]. Clinical
studies have demonstrated the effectiveness of BRD-4 inhibitors in glioma, where BRD-4
expression is significantly higher than in adjacent normal brain tissue [9]. The development of

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these inhibitors has traditionally focused on bromodomains, specifically bromodomain 1 and

bromodomain 2, providing insights into potential therapeutic strategies [8].
The objective of the paper is to explore the molecular mechanisms underlying BRD-4
inhibition, focusing on how inhibitors interact with BRD-4 and modulate its activity. Utilize
structural and biochemical studies to provide detailed insights into the binding interactions.
2. Methodology:
2.1 SMILES Collection: The SMILES notation provided a compact representation of the
chemical structure, and the IC50 values has given an indication of each ligand's potency as an
inhibitor, with lower values suggesting higher potency. Researchers have used this information
to assess the effectiveness of compounds in inhibiting a specific target, in this case, BRD4.
Sr. LIGAND Smiles IC50nM
No ID [10]

1 SQ-9 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=C(F)C=C3)=O)=CC1=O 80.54

2 SQ-17 CN1C2=CC=CC=C2C(OCCCC(NC3CCCCCC3)=O)=CC1=O 95.88
3 SQ-12 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=C(F)C=C3F)=O)=CC1=O 96.33
4 SQ-14 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=CC=C3F)=O)=CC1=O 152.8
5 SQ-19 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=C(C(C)C)C=C3)=O)=CC1=O 254.1
6 SQ-18 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=C(CC)C=C3)=O)=CC1=O 392.6
7 SQ-21 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=CC(C#N)=C3)=O)=CC1=O 585.5
8 SQ-13 CN1C2=CC=CC=C2C(OCCCC(NC3=CC(Cl)=CC=C3OC)=O)=CC1=O 623.8
9 SQ-15 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=CC(OC)=C3)=O)=CC1=O 630
10 SQ-1 O=C3/C=C(/OCCCC(=O)NC=1/C=C(/[Cl])C=CC=1)C2=CC=CC=C2N3C 676.1
11 SQ-10 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=CC(C(F)(F)F)=C3)=O)=CC1=O 1175
12 SQ-11 CN1C2=CC=CC=C2C(OCCCC(NC3=CC=C(C)C=C3Br)=O)=CC1=O 1177
13 SQ-20 CN1C2=CC=CC=C2C(OCCCC(NC3=C(OC)C=C(Cl)C(C)=C3)=O)=CC1=O 1256
14 SQ-8 CN1C2=CC=CC=C2C(OCCCC(NCC3=C(Cl)C=C(F)C=C3)=O)=CC1=O 1650
15 SQ-7 CN1C2=CC=CC=C2C(OCCCC(NCC3=CC=C(F)C=C3)=O)=CC1=O 1688
16 SQ-16 CN1C2=CC=CC=C2C(OCCCC(NC3=CC(Cl)=C(OC)C=C3OC)=O)=CC1=O 1781
17 SQ-5 CN1C2=CC=CC=C2C(OCC(NC3=CC=C(F)C=C3F)=O)=CC1=O 4747
18 SQ-3 CN1C2=CC=CC=C2C(OCC(NC3=CC=C(C)C=C3Br)=O)=CC1=O 5996
19 SQ-4 CN1C2=CC=CC=C2C(OCC(NC3=CC=C(C)C(F)=C3)=O)=CC1=O 9209
20 SQ-6 CN1C2=CC=CC=C2C(OCC(NC3=CC=C(Br)C=C3C)=O)=CC1=O 9307
21 SQ-2 CN1C2=CC=CC=C2C(OCC(NC3=CC=C(F)C=C3)=O)=CC1=O 25748
Note: - Not Determined [10]
Database Selection: Chose a reputable chemical data from literature [10] to collect SMILES
notations of target molecules. Data Filtering: Filtered the dataset based on specific criteria,
such as biological activity or structural features, to ensure relevance to our study.
2.2. SMILES to 3D Conversion:
Software Selection: Used molecular modeling software like RDKit, Open Babel, or Pybel to
convert SMILES to 3D structures using MMFF94 force field. Conversion Process: Utilized
the chosen software to convert SMILES notations into 3D molecular structures, ensuring
proper handling of stereochemistry and atom coordinates.
2.3. Optimization: Geometry Optimization: Performed energy minimization and geometry
optimization on the 3D structures to obtain stable conformations. Tools like RDKit and Open
Babel provided optimization functionalities.
2.4. Saving as .SDF File:
File Format Conversion: Saved the optimized 3D structures in the Structure Data File (.sdf)
format. Most molecular modeling software tools support this format for saving molecular

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information. Consistency Check: Ensured that the saved .sdf file maintains the integrity and
consistency of the molecular structures.
2.5. Pharmacophore Generation using PyMOL and Liquid Plugin: Software Installation:
Executed PyMOL, a powerful molecular visualization tool, and the Liquid plugin for
pharmacophore generation. Opened the saved .sdf file containing optimized structures in
PyMOL.
Pharmacophore Generation: Used the Liquid plugin to generate pharmacophores based on
specific features such as hydrogen bond donors/acceptors, hydrophobic regions, and aromatic
centers. Visualization: Visualized the generated pharmacophores in PyMOL for further
analysis.

3. Results and Discussion:

Our research represents a significant stride forward in the pursuit of enhanced therapeutics
for prostate cancer. The systematic identification and optimization of BRD4 inhibitors,
underscored by the innovative integration of pharmacophore modeling, have collectively
marked a breakthrough in precision medicine for cancer treatment. The tailored design and
optimization of these inhibitors exhibit a promising avenue for addressing the unique
challenges posed by prostate cancer.
Through a meticulous exploration of molecular interactions and structural features, this
study provides valuable insights that contribute to the refinement of BRD4 inhibitors. The
potential impact of these findings extends beyond the laboratory, offering a blueprint for the
development of targeted therapies in the clinical setting. The fusion of cutting-edge drug design
strategies with computational modeling techniques not only breaks new ground but also lays a
foundation for the evolution of personalized treatment approaches.
As we move forward, the outcomes of this research hold the potential to reshape the
landscape of prostate cancer therapeutics. By breaking new ground in the identification and
optimization of BRD4 inhibitors and harnessing the power of pharmacophore modeling, we
chart a course towards more effective and tailored interventions. This study serves as a catalyst
for future advancements, fostering a paradigm shift in the way we approach precision medicine
for prostate cancer and potentially other malignancies.

SQ-9 SQ-2
Most active Least active
Figure3.1: - 3D Structure of most active and least active molecules, with distorted geometry
of molecules.

SQ-9 SQ-2
Most active Least active

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Figure3.2: - 2D Structure of most active and least active molecules.

SQ-9 SQ-2
Most active molecule Least Active molecule
Figure3.3: - Above figure shows the pharmacophore model structure of most active and least
active molecules.

Pharmacophore modelling green colour for lipophilic, blue colour for H-donor and red
colour for H-acceptor. In this figure SQ-2 structure shows three lipophilic, four H-acceptor and
one H-donor molecules. In SQ-9 structure four lipophilic, four acceptor and one donor
molecules.

SQ-9 SQ-2
Figure 3.4: - In this above structure SQ-9 and SQ-2 both showing Same Number of H-
acceptor

SQ-9 SQ-2
Figure 3.5: - SQ-9 having four lipophilic parts and SQ-2 having only three lipophilic part in
this figure.

SQ-9 SQ-2
Figure 3.6: - In this above structure SQ-9 and SQ-2 both showing Same Number of H-donor
(one H-Donor)

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SQ-9 SQ-2
Figure3.7: - Measure distances between Lipophilic, H-acceptor and H-donor in most active
and least active molecules.
In the SQ-9 shows distorted geometry with four lipophilic parts, the lipophilic to lipophilic
higher distance is 10.9A0 and lowest distance is 4.9A0, H-acceptor to H-acceptor distance is
4.2 A0, H-acceptor-Lipophilic distance is 2.8 A0 and H-acceptor to H-donor distance is 8.8A0.
The SQ-2 having only three lipophilic parts in the molecule, in this structure lipophilic-
lipophilic higher distance is 11.0A0 and lipophilic to lipophilic lowest distance is 4.9A0, H-
acceptor to H-acceptor distance is 2.5 A0, H-acceptor to Lipophilic distance is 2.8 A0, and H-
acceptor to H-donor distance is 3.7A0.

Conclusion:-
In conclusion, our research represents a significant stride forward in the pursuit of enhanced
therapeutics for prostate cancer. The systematic identification and optimization of BRD4
inhibitors, underscored by the innovative integration of pharmacophore modeling, have
collectively marked a breakthrough in precision medicine for cancer treatment. The tailored
design and optimization of these inhibitors exhibit a promising avenue for addressing the
unique challenges posed by prostate cancer. Through a meticulous exploration of molecular
interactions and structural features, this study provides valuable insights that contribute to the
refinement of BRD4 inhibitors. The potential impact of these findings extends beyond the
laboratory, offering a blueprint for the development of targeted therapies in the clinical setting.
The fusion of cutting-edge drug design strategies with computational modeling techniques not
only breaks new ground but also lays a foundation for the evolution of personalized treatment
approaches.
As we move forward, the outcomes of this research hold the potential to reshape the landscape
of prostate cancer therapeutics. By breaking new ground in the identification and optimization
of BRD4 inhibitors and harnessing the power of pharmacophore modeling, we chart a course
towards more effective and tailored interventions. This study serves as a catalyst for future
advancements, fostering a paradigm shift in the way we approach precision medicine for
prostate cancer and, potentially, other malignancies.
"Sincere acknowledgement to Prof. Dr. Vijay H. Masand for his support throught
the research work."

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