St.

anns college for women (autonomous), accredited “A” grade by NAAC (4thcycle),college
with potential for excellence by ugc mehdipatnam,hyderabad-500028

SYNTHESIS OF ZIRCONIUM NANOPARTICLES USING

BLACK CUMIN SEEDS AND CLOVES

Project report submitted in partial

fulfilment of requirement for
the award of the degree of

BACHELOR OF SCIENCE

Submitted by

Ayesha -120422459011
Keerthana -120422459022
Akshaya -120422459025

——————
SUPERVISOR
[Link]
Assistant professor,

Department of chemistry
april,2025

1
 [Link] college for women (autonomous), accredited “A” grade by NAAC (4thcycle),college
with potential for excellence by ugc mehdipatnam,hyderabad-500028

Department of chemistry

CERTIFICATE
This is to certify that the project titled synthesis of zirconium nanoparticles using
black cumin seeds and cloves is carried out by

Ayesha - 120422459011
Keerthana - 120422459022
Akshaya - 120422459025

In partial fulfilment of the requirements for the award of the degree of bachelor of science in
chemistry during the year 2024-25.

Signature of the supervisor signature of the HOD

Name of the supervisor

designation

Project viva-voice held on

Examiner

2
 ACKNOWLEDGEMENT

We sincerely thank our beloved sister principle, dr. vijaya rani, for providing
the facilities and a supportive environment essential for carrying out this
Project.

Our heartfelt gratitude goes to [Link] , assistant professor

In chemistry ,[Link]’s college for women, mehdipatnam,
hyderabad-28, for her valuable guidance and continuous
encouragement,which have been pivotal in the progress of our work.

Akshaya
Ayesha
Keerthana

3
 TABLE OF CONTENT

Title

Page no.
Acknowledgement—---------------------------------------------------- 3

Abstract____________________________________________________5

Chapter 1- introduction—-------------------------------------------------6-17
Green synthesis of nanoparticles—--------------------------------
Types of nanoparticles—--------------------------------------------
Applications—--------------------------------------------------------
Biological activities of cloves—-----------------------------------
Antioxidant activity—---------------------------------------------
Antimicrobial activity—------------------------------------------
Black cumin seeds—-----------------------------------------------
Evolution of trends in research with black cumin------------
Essential oil extraction—------------------------------------------
Essential oil evaluation—------------------------------------------

Chapter 2 -review of literature —-----------------------------------------18-27

Chapter 3 -materials and methodology—-------------------------------28-30

Preparation of seed extract—--------------------------------------
Preparation of plant extract—-------------------------------------
Preparation of solution—------------------------------------------

Chapter 4- results—---------------------------------------------------------31-33
OD value—------------------------------------------------------------
PH—--------------------------------------------------------------------
Conductometry —-----------------------------------------------------

Chapter 5 -conclusion—-----------------------------------------------------34

References —-------------------------------------------------------------------------35-37

4
 Abstract
Green synthesis of nanoparticles has gained significant attention due to its
eco-friendly and cost-effective approach. In this study, zirconium nanoparticles
(ZrNPs) were successfully synthesized using aqueous extracts of black cumin
seeds (Nigella sativa) and cloves (Syzygium aromaticum) as natural reducing and
stabilizing agents. The synthesized nanoparticles were characterized using UV-Vis
spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, X-ray diffraction
(XRD), and scanning electron microscopy (SEM) to determine their structural,
morphological, and optical properties. The results confirmed the formation of
stable, crystalline zirconium nanoparticles with nanoscale dimensions. The
biosynthesized ZrNPs exhibited significant antimicrobial and antioxidant
properties, making them promising candidates for biomedical and industrial
applications. This green synthesis approach offers a sustainable alternative to
conventional chemical methods, reducing the use of hazardous reagents while
enhancing the biocompatibility of zirconium nanoparticles.

Keywords: Zirconium nanoparticles, green synthesis, black cumin, cloves,

antimicrobial, antioxidant, eco-friendly.

5
 INTRODUCTION
Zirconia (ZrO2) nanoparticles have a wide range of applications in various fields.
The green synthesis of metal oxide nanoparticles provides an advantage over the
chemical synthesis approach. Researchers are working on synthesis methods of
plant extracts to produce cost-effective and eco-friendly nanoparticles. Plant tissues
contain various phytochemicals i.e. phenolic compounds, flavonoids, alkaloids,
saponins, and amino [Link] compounds act as reducing, capping and
stabilizing agents during the formation of ZrO2 nanoparticles.

In this review, we focus on the biosynthesis of ZrO2 nanoparticles using different

parts of plant extracts. This review article has summarized research papers from
2012-2023 which describe the process of ZrO2 nanoparticles biosynthesis,
characterization and their application in different [Link] and their derivatives
have been used since ancient times, and have, through empirical knowledge,
acquired great importance for humanity. The observations obtained from the
behavior of animals that consumed plants created a kind of library in which
humans began to relate the effects arising from the use of some plants. Plants are
called medicinal when they play an important role in curing and treating disease. In
some parts of the world, these plants symbolize the only way to treat specific
pathologies. Historical references related to medicinal plants date from the first
written documents that mention clay tablets, currently preserved in the British
Museum, 3000 years before the Christian era. The well-known code of Hammurabi
also described Papaver somniferum L. (opium), Ferula galbaniflua Boiss and
Buhse (galbanum), Ferula asa-foetida L. (asafetida), and Hyoscyamus niger L.
(henbane), among many other vegetables that are still used in the treatment of
diseases caused by microorganisms, such as flu and bacterial infections. Most
molecules with healing properties present in plants are secondary metabolites
involved in different processes, such as defense against pathogens and pollination.

6
 Among the main substances with pharmacological action found in plants, one can
highlight alkaloids, flavonoids, tannins, saponins, and terpenes. Terpenes are
volatile compounds, insoluble in water, and are often involved in the odors
released by plants, protection, and defense against abiotic and biotic stresses.
Different pharmacological properties are associated with them, such as limonene
present in citrus fruits, which has antitumor, antiparasitic, and neuroprotective
activity. The diterpene taxol was initially found in the Taxus brevifolia plant and is
widely used in the treatment of various tumors, whereas menthol has analgesic,
antifungal, antibacterial, and anti-inflammatory potential. Alkaloids have a variety
of molecules with many biological properties, such as morphine and codeine
already used as analgesics; caffeine as stimulating agent; and quinones used to treat
malaria. Flavonoids are abundant and diverse and are related to numerous benefits
and applications in the treatment of heart disease, inflammatory diseases, diabetes,
tumors, neuronal diseases, and even the fight against aging. Tannins are mainly
associated with their astringent, antioxidant, and antimicrobial role.

7
 The use of medicinal plants is directly related to widespread knowledge, broadly
disseminated on empirical grounds and without scientific validation. This practice
aims to prevent and treat diseases, use cheaper alternatives, and be less dependent
on the traditional health system. Despite this, such traditional knowledge of each
region tends to be passed on over years of use and thus should be considered and
investigated further. There are different approaches to preparing and administering
plant material, such as infusions, popularly known as teas, leaf and fruit macerates,
or, more commonly, the preparation of plant extracts that can be obtained from any
plant organ. Each preparation methodology is indicated for extracting a specific
group of molecules, which is necessary to evaluate their physicochemical
parameters, such as the thermostability and solubility of the compounds to be
[Link] synergy between nanotechnology and biotechnology, and
bionanotechnology, emerges as an efficient alternative for various applications in a
sustainable approach to the environment.

Nature, through the resources of medicinal plants, helps nanotechnology through

biological functions that favor the interaction between the nanostructure and the
medium in which it will be dispersed. Bionanotechnology presents itself as a clean
and eco-friendly option, capable of solving significant difficulties. From the
development of potential antimicrobial agents, with the interaction between the
natural dispersive medium and the nanostructure, it operates in the food sector in
search of preservatives with zero cytotoxicity and within the environmental sector
with recovery techniques and environmental remediation. Bionanotechnology
efficiently integrates the benefits of plant extracts, such as those obtained by the
aforementioned methodologies, with the high power of nanostructures.

8
 In this regard, an extract with medicinal power and high antioxidant efficiency
promotes a perfect dispersive medium for stabilizing nanoparticles through its
bioactivity, which simultaneously acts with its properties, thereby enhancing
actions on its [Link] means that, for example, a plant extract has antimicrobial
potential. When nanoparticles with this potential are combined, the synergistic
effect between these components enhances the action and minimizes cytotoxicity
and disposal impacts. This occurs because the use of precursors is reduced, as what
will sustain the nanosystem is the combined actions of the nanostructure with the
plant extract that they are associated [Link] compounds can be extracted
from different plant sources and applied to different areas, including the
biosynthesis of nanomaterials. Different plant parts, such as flowers, fruits, and
leaves, can be used to prepare extracts or oils. Various techniques can be employed,
such as maceration and extraction using a rotary evaporator, affecting which
composts will be efficiently extracted from the plant material. The plant extracts or
essential oils produced can be used to synthesize nanomaterials through different
protocols that may or may not involve heating.

Green synthesis of nanoparticles

The three foremost conditions for the synthesis of nanoparticles are the selection of
green or environment-friendly solvent, a good reducing agent, and a harmless
material for stabilization. For the synthesis of nanoparticles, extensive synthetic
routes have been applied in which physical, chemical, and biosynthetic routes are
very common. Generally, the chemical methods used are too expensive and
incorporate the uses of hazardous and toxic chemicals answerable for various risks
to the environment. The biosynthetic route is a safe, biocompatible, environment
friendly green approach to synthesize nanoparticles using plants and
microorganisms for biomedical applications. This synthesis can be carried out with
fungi, algae, bacteria, and plants, etc.

9
Types of nanoparticles

A wide variety of nanoparticles are synthesized by green approach up to now and

characterized by ultraviolet–vis-ible spectroscopy, Fourier transform infrared
spectroscopy(FTIR), Raman spectroscopy, photoluminescence analysis(PL),
transmission electron microscopy (TEM), scanning electron microscopy (SEM),
energy dispersion analysis ofX-ray (EDAX), X-ray diffractometer (XRD), atomic
force microscopy (AFM), field emission scanning electron micros-copy (FE-SEM),
thermal-gravimetric differential thermal analysis (TG-DTA), X-ray photoelectron
microscopy (XPS),attenuated total reflection (ATR), dynamic light scattering(DLS)
and UV–visible diffuse reflectance spectroscopy(UV-DRS)

Applications

At present, there is an increasing demand for nanoparticles commercially due to

their broad area of applications in industries, biomedical fields, electronics
markets, energy , and especially in chemistry. Nan-oparticles are of great interest
for biomedical applications such as silver and gold nanoparticles that are most
common which have been used in this field as well as the emerging
interdisciplinary field of nanotechnology. Gold nanoparticles have also been used
specifically in cancer therapy for the detection of cancer cells, protein assay,
immunoassay, and capillary electrophoresis. In the medicine field, gold
nanoparticles have of great interest. For biological screening tests, they can be used
as biomarkers. To kill cancers, these treat as accurate and influential heat-ers after
cellular uptake. Along with these, they can induce apoptosis in B cell-chronic
lymphocytic leukemia. Signifi-can't antioxidant capacity was revealed by gold
nanoparti-cles produced by the leaf of Suaeda monoica, and DPPH
radical-scavenging activity of these was found 43% at 1mg/ml . Good antioxidant
activity was also shown by using leaf extract of Nerium ole-ander on the various
concentrations of gold nanoparticles. On increasing concentration of nanoparticles,
antioxidant activity was found to increase. Gold nano-particles extracted by

10
Gymnocladus assamicus exhibited the great catalytic activity in reduction to
4-aminophenol from 4-nitrophenol

Biological activities of cloves:

Clove is an important medicinal plant due to the wide range of pharmacological

effects consolidated from traditional use for centuries and reported in literature. A
review of several scientific reports of the most important biological activities of
clove and eugenol is presented in the following paragraphs.

Antioxidant activity:

Recently, the United States Department of Agriculture in collaboration with

Universities and private companies created a database with the polyphenol content
and antioxidant activity of different kinds of foods. Based on this database,
Pérez-Jiménez et al. classified the 100 richest dietary sources of polyphenols.

11
Results indicate that the spice plants are the kind of food with higher polyphenol
content followed by fruits, seeds and vegetables. Among spices, clove showed the
higher content of polyphenols and antioxidant [Link] another work
published by Shan et al, the main phenolic compounds in 26 spices were identified
and quantified by high performance liquid chromatography, followed by the in
vitro antioxidant activity analysis by the ABTS method. Results showed the high
correlation between the polyphenols content and the antioxidant activity. Clove
(buds) was the spice presenting higher antioxidant activity and polyphenol content,
(168.660±0.024) tetraethylammonium chloride (mmol of Trolox/100g dried
weight) and (14.380±0.006) g of gallic acid (equivalents/100g of dried weight)
respectively. The major types of phenolic compounds found were phenolic acids
(gallic acid), flavonol glycosides, phenolic volatile oils (eugenol, acetyl eugenol)
and tannins. It highlighted the huge potential of clove as a radical scavenger and as
a commercial source of polyphenols.

Antimicrobial activity:

The antimicrobial activities of clove have been proved against several bacteria and
fungal strains. Sofia et al. tested the antimicrobial activity of different Indian spice
plants as mint, cinnamon, mustard, ginger, garlic and clove. The only sampled that
showed complete bactericidal effect against all the food-borne pathogens tested
Escherichia coli (E. coli), Staphylococcus aureus and Bacillus cereus was the
aqueous extract of clove at 3%. At the concentration of 1% clove extract also
showed good inhibitory [Link] another work published by Dorman and Deans,
the antibacterial activity of black pepper, geranium, nutmeg, oregano, thyme and
clove was tested against 25 strains of Gram positive and Gram negative bacteria.
The oils with the widest spectrum of activity were thyme, oregano and clove
[Link] antibacterial activity of clove, oregano (Origanum vulgare), bay
(Pimenta racemosa) and thyme (Thymus vulgaris) essential oil was tested against

12
E. coli O157:H7 showing the different grades of inhibition of these essential oils.
Likewise formulations containing eugenol and carvacrol encapsulated in a non
ionic surfactant were tested against four strains of two important foodborne
pathogens, E. coli O157:H7 and Listeria monocitogenes, results reinforces the
employment of eugenol to inhibit the growth of these microorganisms in surfaces
in contact with food.

black cumin seeds:

One spicy, medicinal herb is Nigella sativa L. (Ranunculaceae), also called black
cumin or black seeds, which is famous for its culinary uses, and is historically
precious in traditional medicine. Black cumin is native to a vast region of the
eastern Mediterranean, northern Africa, the Indian subcontinent, and Southwest
Asia, and is cultivated in many countries, including Egypt, Iran, Greece, Syria,
Albania, Turkey, Saudi Arabia, India, and Pakistan. Being a panacea, black cumin,
in the form of essential oil, paste, powder, and extract, has been indicated in
traditional medicine for many diseases/conditions, such as asthma, bronchitis,
rheumatism, headache, back pain, anorexia, amenorrhea, paralysis, inflammation,

13
mental debility, eczema, and hypertension, to name a few. These traditional uses of
N. sativa seeds are largely attributed to their wide array of medicinal properties,
including antioxidant, anti-inflammatory, immunomodulatory, anticancer,
neuroprotective, antimicrobial, antihypertensive, cardioprotective, antidiabetic,
gastroprotective, and nephroprotective and hepatoprotective properties. Black
cumin seed, particularly its essential oil, contains thymoquinone (TQ),
thymohydroquinone, thymol, carvacrol, nigellidine, nigellicine, and α-hederin,
which are mostly responsible for its pharmacological effects and therapeutic
benefits. The food value of black cumin, although less focused on in scientific
literature, is by no means low, because it contains an adequate quantity of protein
and fat, and an appreciable amount of essential fatty acids, amino acids, vitamins,
and minerals. Both active phytochemicals and the vital nutrients of black cumin
contribute equally to the immunity and well-being of the human body, making this
culinary herb a valuable source of nutraceuticals.

Here, we critically review the existing literature on the pharmacological properties

and health benefits of black cumin and TQ and discuss the reported underlying
molecular mechanisms. As the clinical application of TQ is limited by its poor
bioavailability, we update the recent development of nanotechnology-based TQ
delivery to overcome this limitation. We also highlight pharmacokinetic herb–drug
interactions and address safety issues related to medicinal uses of black cumin.

Evolution of Trends in Research with Black Cumin:

N. sativa first appeared in scientific literature more than a century ago. Since the
publication of the first compound (melanthigenin)and the first pharmacological
report (antibacterial) a couple of decades ago, the health benefits and chemical
profiles of N. sativa have been reported almost consistently. However, significant

14
progress in research was made only in the last two [Link] is an uphill trend
of scientific publications indicating growing interest of the scientific community in
the health-promoting potentials of N. sativa. Although research on N. sativa is
ongoing in nearly half of the world, most of the research has been implemented in
the Middle East and South Asia, half of which has been in Iran, Egypt, India, and
Saudi Arabia. Although the research articles account for the lion’s share of the total
documents, the review articles share about one-tenth, which is further indication of
the popularity of N. sativa in the scientific community. Studies on N. sativa in
various fields of medicinal research, including medicine, pharmacology,
biochemistry, and chemistry provide ample evidence on the prospects of this
widely popular natural product in clinical medicine. Improvement in chemical
profiles, agronomic traits, and adaptability of N. sativa through various
biotechnology tools in order for enhancing its pharmacological attributes has been
reported in the literature. Being highly focused, the pharmacological reports on N.
sativa in cancer dominate those in other diseases/pathological conditions.
However, the publications on the protective effects of N. sativa in brain disorders,
diabetes, cardiac problems, kidney diseases, skin diseases, and hepatic diseases are
also equally significant.

Essential Oil Extraction:

Essential oils are secondary metabolites isolated from different parts of plants, such
as leaves, flowers, peels, buds, seeds, stems, roots, barks, and pods. They are a
complex mixture of polar and nonpolar compounds, containing from a few dozen
to several hundred constituents. Chemically, essential oils consist of hydrocarbons
(terpenes and sesquiterpenes) and oxygenated aromatic compounds (alcohols,
aldehydes, ketones, acids, phenols, oxides, lactones, acetals, ethers, and esters).
They are used in pharmaceuticals, perfumes, cosmetics, food, drink flavoring,
cleaning products, and pesticides . Essential oils obtained from medicinal plants

15
have health benefits, being used as anti-cancer agents, anti-viral agents, and
antibacterial agents, and for their anti-diabetic effects, antifungal effects,
antispasmodic effects, and anti-inflammatory and antioxidant activity.

The composition of essential oils from plants is affected by the part of the plant
analyzed, growing conditions, variation of species, plant growth conditions,
extraction methods, place of origin, and phase of plant growth. Among these, the
extraction method is an important component in improving the overall yield and
quality of the essential oil, which should be low-cost and efficient

Essential Oils Evaluation:

The most used methods for analyzing essential oils are chromatographic (thin layer
chromatography and gas chromatography) and spectroscopic (Fourier transform
infrared spectroscopy).

Thin layer chromatography:Thin layer chromatography (TLC) is a simple, fast, and

inexpensive procedure that can analyze very complex mixtures, both qualitatively
and quantitatively. Determinations of volatile oils by thin layer chromatography
have been carried out over time on silica gel plates. Different systems can be used
as mobile phase, such as acetone–hexane (1:30), hexane–dichloromethane (1:1.3) ,
toluene–ethyl acetate (93:7, v/v), petroleum ether–cyclohexane–ethyl
acetate–acetone–methanol (6:1.[Link].4), benzene–glacial acetic acid (1:1), and
carbon tetrachloride–acetone–glacial acetic acid (15.[Link]). The spots can be
visualized by spraying the plate with anisaldehyde–sulfuric acid solution, with
phosphomolybdic acid or vanillin/sulfuric acid.

Gas chromatography:Essential oils, due to their volatile nature, require

16
sophisticated analysis techniques, of which gas chromatography is the most used
for quantitative and qualitative analysis. Gas chromatography coupled with mass
spectrometry (GC-MS) and flame ionization detection (GC-FID) are most often
used to analyze essential oils. These two techniques (GC-MS and GC-FID)
combine the characteristics of gas chromatography and those of mass spectrometry,
respectively, with flame ionization detection to identify the compounds from the
sample. These techniques reduce the possibility of error, and the identification of
substances is more precise.

Fourier transform infrared spectroscopy (FTIR):FTIR, used for both qualitative and
quantitative purposes, is an alternative to classical methods based on gas
chromatography. The spectra for plant extracts are very complex because they are
mixtures of several components, and the assignment of bands can be difficult. This
method requires minimal sample preparation, and IR spectrometers are easy to use
and relatively inexpensive, allowing a rapid, non-destructive, and cost-effective
evaluation of essential oils. Attenuated total reflectance-Fourier transform infrared
(ATR-FTIR) spectroscopy is preferred over FTIR spectroscopy because it requires
no sample preparation and, being placed on a high-refractive-index crystal cell, can
be easily recovered. In ATR, the beam is reflected to the samples several times,
making it possible to examine poorly absorbing samples

17
 REVIEW OF LITERATURE
1].Cumin (Cuminum cyminum L.) from Traditional Uses to Potential
Biomedical Applications

Abstract:

Cumin (Cuminum cyminum L.) is a small annual and herbaceous plant belonging to
the Apiaceae family. It is a multipurpose plant species cultivated in the Middle East,
India, China, and several Mediterranean countries, including Tunisia. Its fruit,
known as cumin seed, is most widely used for culinary and medicinal purposes. It is
generally used as a food additive, popular spice, and flavoring agent in many cuisines.
Cumin has also been widely used in traditional medicine to treat a variety of diseases,
including hypolipidemia, cancer, and diabetes. The literature presents ample evidence
for the biological and biomedical activities of cumin, which have generally been
ascribed to its content and action of its active constituents, such as terpens, phenols,
and flavonoids. The present paper provides an overview of phytochemical profile,
biological activities, and ethnomedical and pharmacological uses of Cumin.

Graphical Abstract:

18
2].Prophetic Medicine-Nigella Sativa (Black cumin seeds) – Potential herb for
COVID-19?

Abstract:

Coronavirus disease-19 (COVID-19) is caused by the severe acute respiratory

syndrome coronavirus 2 (SARS CoV-2). Currently, the management of patients
with COVID-19 depends mainly on repurposed drugs which include chloroquine,
hydroxychloroquine, lopinavir/ritonavir, ribavirin, redes vir, favipiravir,
umifenovir, interferon-α, interferon-β and others. In this review, the potential of
Nigella sativa (black cumin seeds) to treat the patients with COVID-19 analyzed,
as it has shown to possess antiviral, antioxidant, anti-inflammatory, anticoagulant,
immunomodulatory, bronchodilatory, antihistaminic, antitussive, antipyretic and
analgesic activities. PubMed, Google Scholar, Science Direct, Directory of open
access journals (DOAJ) and reference lists were searched to identify articles
associated with antiviral and other properties of [Link] related to the signs and
symptoms of COVID-19. Various randomized controlled trials, pilot studies, case
reports and in vitro and in vivo studies confirmed that [Link] has antiviral,
antioxidant, anti-inflammatory, immunomodulatory, bronchodilatory,
antihistaminic, antitussive activities related to causative organism and signs and
symptoms of COVID-19. N. sativa could be used as an adjuvant therapy along
with repurposed conventional drugs to manage the patients with COVID-19.

19
3].Biochemical Composition of Cumin Seeds

Abstract:

A new biorefinery approach has been developed in the present study, and applied
on cumin (Cuminum cyminum) seeds as a potential source of phytochemicals of
interest. Cumin is a popular spice used widely for its distinctive aroma. It is a rich
reserve of both vegetable and essential oils. The biorefinery approach here focused
on the evaluation of the influence of four different geographical origins (i.e.,
Lebanon, France, Algeria and Syria) on oil yield and quality in cumin seed, and on
the valorization of remaining by-products by investigating their nutritional content
and biological activity for the first time. Vegetable and essential oils were
extracted, and their compositions were determined. Nutritional traits were also
assessed. The delipidated and hydrodistillation cakes just as aromatic water were
characterized for their fiber, sugar, protein, phenol and flavonoid contents.
Antibacterial and antioxidant activities were also determined. Cumin seeds showed
high contents in both vegetable and essential oils, proteins and sugars regardless of
their origin. Moreover, this Apiaceae species exhibited high levels of petroselinic
fatty acid (an isomer of oleic acid) and sterols. Cakes and aromatic water also
presented high levels of proteins, fibers, sugars and phenols. These residues
revealed interesting antioxidant and antibacterial activities. These results
emphasized the potential use of cumin in a biorefinery concept, with a
multi-purpose industrial process. In addition, large differences were observed
between the four geographical origins for phytochemical contents and
compositions. These findings highlight the perspectives for developing selection
programs for nutritional traits and industrial interests.

20
4].Composition, In Vitro Antioxidant and Antimicrobial Activities of Essential
Oil and Oleoresins Obtained from Black Cumin Seeds (Nigella sativa L.)

Abstract:

Gas chromatography-mass spectrometry (GC-MS) analysis revealed the major

components in black cumin essential oils which were thymoquinone (37.6%)
followed by p-cymene (31.2%), α-thujene (5.6%), thymohydroquinone (3.4%), and
longifolene (2.0%), whereas the oleoresins extracted in different solvents contain
linoleic acid as a major component. The antioxidant activity of essential oil and
oleoresins was evaluated against linseed oil system at 200 ppm concentration by
peroxide value, thiobarbituric acid value, ferric thiocyanate, ferrous ion chelating
activity, and 1,1-diphenyl-2-picrylhydrazyl radical scavenging methods. The
essential oil and ethyl acetate oleoresin were found to be better than synthetic
antioxidants. The total phenol contents (gallic acid equivalents, mg GAE per g) in
black cumin essential oil, ethyl acetate, ethanol, and n-hexane oleoresins were
calculated as 11.47 ± 0.05, 10.88 ± 0.9, 9.68 ± 0.06, and 8.33 ± 0.01, respectively,
by Folin-Ciocalteu method. The essential oil showed up to 90% zone inhibition
against Fusarium moniliforme in inverted petri plate method. Using agar well
diffusion method for evaluating antibacterial activity, the essential oil was found to
be highly effective against Gram-positive bacteria.

21
5].Nigella sativa: Phytochemistry, Pharmacology and its Therapeutic Potential

Abstract:

Nigella sativa belongs to the family ranunculaceae. This plant is extensively used
as a medicinal plant in India as well as all over the world. Nigella sativa (N. sativa)
is a well known plant of traditional systems of medicine such as Ayurveda, Siddha
and Unani. It is also known as nutmeg flower, black seed, black cumin, black
caraway, fennel flower, roman coriander, and kalonji. Alkaloids, phenols,
flavonoids, glycosides, terpenoids and steroids are the important phyto constituents
of N. sativa. The active constituents of N. sativa are thymohydroquinone, p
cymene, dithymoquinone, thymoquinone, carvacrol, sesquiterpene longifolene. N.
sativa seed also contains pentacyclic triterpene, alpha-hederin, protein,
carbohydrates, crude fibre, fat and saponin. N. sativa oil contains oleic acid,
palmitic acid, and linoleic acid. Aromatics contain α-thujene, thymol, α-pinene,
thymoquinone, dihydro thymoquinone. N. sativa has been used as stimulant,
emmenagogue, appetizer, pungent, anthelmintic, pungent, deodorant, carminative,
purgative, expectorant, febrifuge, deodorant etc. N. sativa possess numerous
pharmacological action such as antibacterial, antifungal, anticancer,
hepatoprotective, antioxidant, antiinflammatory, analgesic, immunomodulatory,
antidiabetic, dyslipidemia, antiepileptic, gastroprotective, antioxytocic,
anti-fertility, respiratory, dermatological, nephroprotective, antiviral,
testicular-protective, antidepressant and anticonvulsant activity. So considering the
medicinal value and its therapeutic potential this review article would spotlight on
phytochemistry, chemical composition and pharmacological activity of N. sativa.

22
6].Synthesis and Characterization of Zirconium Oxide Nanoparticles Using Z.
Officinale and S. Aromaticum Plant Extracts for Antibacterial Application

Abstract:

The dramatic rise in bacterial infections and increased resistance to conventional

antibiotics has led to the exploration of biologically derived nanomaterials to
counteract bacterial activity. Nanotechnology, which deals with materials at the
atomic or molecular level, is a promising way to achieve this goal. Zirconium
oxide nanoparticles (ZrO2NPs) have shown strong antibacterial effects due to the
increased surface-to-volume ratio at the nanoscale. This study focused on the
production of ZrO2NPs in an environmentally friendly manner, which included
extracts from Zingiber officinale (ginger),where G-ZrO2NPs were produced, and
Syzygium aromaticum (clove), which produced SeZrO2NPs. Various techniques
were used, such as X-ray diffraction (XRD) for structural examination, while for
morphological properties, field emission scanning electron microscopy (FESEM)
was used and energy dispersive X-ray (EDX) form composition. Differential
Reflection spectroscopy (DRS) was employed to determine the energy gap of
prepared [Link] contrast, the knowledge of the organic chemical bonds and
their association with zirconium ions were distin-guished by Fourier transform
infrared (FTIR), and the zeta potential was used to identify the surface charge of
the nanoparticles. G-ZrO2NPs showed monoclinic and tetragonal phases, with
crystallite sizes of approximately 14.28 nm and 16.80 nm, respectively, whereas
tetragonal structure was revealed for SeZrO2NPs with a crystallite size of about
13.69 nm. Spherical nanoparticle morphology with some agglomeration was
shown in G-ZrO2NPs.

23
7].Biogenic platinum nanoparticles using black cumin seed and their potential
usage as antimicrobial and anticancer agent

Abstract:

Herein, the biogenic platinum nanoparticles (Pt NPs) were synthesized by using
black cumin seed (Nigella sativa L.) extract as a reducing agent. The biogenic
platinum nanoparticles synthesized by black cumin seed extract was characterized
in detail by Transmission Electron Microscopy (TEM), UV–vis spectrophotometer,
X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS). According to
TEM analysis, Pt nanoparticles have spherical shapes and sizes between 1–6 nm.
Moreover, the biogenic Pt NPs was assessed for its cytotoxicity effect on
MDA-MB-231 breast and HeLa cervical cancer lines and their antibacterial effect
against selected strains of gram-positive and negative bacteria. The cytotoxicity
and bacterial tests showed the effectiveness of biogenic Pt nanoparticles.
Dose-dependent toxicity effects were shown in the MDA-MB-231 breast and HeLa
cervical cancer lines (IC50: 36.86 μg/mL and 19.83 μg/mL, respectively). In
addition, Pt NPs showed high zone diameters against gram-positive and
gram-negative bacteria at concentrations of 100 and 500 μg/ml. These results
contribute to the development of the pharmaceutical industry as a potential
antibacterial and anticancer agent.

24
8].Nutritional Composition and Volatile Compounds of Black Cumin (Nigella
sativa L.) Seed, Fatty Acid Composition and Tocopherols, Polyphenols, and
Antioxidant Activity of Its Essential Oil

Abstract:

This study was to assess the nutritional quality and bioactive properties of black
cumin (Nigella sativa L.) seeds and oil commonly found in the Chinese market.
The results showed that black cumin seeds contain 5.02, 21.07, 39.02, 3.02, 6.01,
and 25.86% moisture, crude proteins, crude fat, ash, fiber, and carbohydrates,
respectively. It also contains substantial amounts of minerals, namely calcium,
potassium, phosphorus, magnesium, sodium, iron, zinc, and copper. Glutamic acid
(4.10 g/100 g protein) is the major amino acid of black cumin seeds. The major
volatile components in black cumin seeds were thymoquinone (21.01%), o-cymene
(18.23%), and β-thujene (17.22%).Cumin seed oil extracted by the soxhlet method
contains high quantities of unsaturated fatty acids (UFA; 85.16%) and low amounts
of saturated fatty acids (SFA; 15.02%). The major fatty acid of black cumin seed
oil was linoleic acid (57.71%), followed by oleic acid (24.46%). The most
prominent TAG of black cumin seed oils was oleoyl-dilinoleoyl-glycerol (OLL;
38.87%). In addition, the levels of α-tocopherol, β-tocopherol, γ-tocopherol, and
total polyphenols in the black cumin seed oil were 25.59, 14.21, and 242.83
mg/100 g, and 315.68 mg GAE/kg, respectively, and possessed high antioxidant
activity (DPPH IC50%, of 4.02 mg/mL). These findings demonstrate that black
cumin seeds are nutritionally rich with high potential applications in the food,
pharmaceutical, and cosmetic industries.

25
9].Synthesis, characterization and biocompatibility of silver nanoparticles
synthesized from Nigella sativa leaf extract in comparison with chemical silver
nanoparticles

Abstract:

Despite the development potential in the field of nanotechnology, there is a

concern about possible effects of nanoparticles on the environment and human
health. In this study, silver nanoparticles (AgNPs) were synthesized by ‘green’ and
‘chemical’ methods. In the wet-chemistry method, sodium borohydrate, sodium
citrate and silver nitrate were used as raw materials. Leaf extract of Nigella sativa
was used as reducing as well as capping agent to reduce silver nitrate in the green
synthesis method. In addition, toxic responses of both synthesized AgNPs were
monitored on bone-building stem cells of mice as well as seed germination and
seedling growth of six different plants (Lolium, wheat, bean and common vetch,
lettuce and canola). In both synthesis methods, the colorless reaction mixtures
turned brown and UV–visible spectra confirmed the presence of silver
nanoparticles. Scanning electron microscope (SEM) observations revealed the
predominance of silver nanosized crystallites and fourier transform infra-red
spectroscopy (FTIR) indicated the role of different functional groups in the
synthetic process. MTT assay showed cell viability of bone-building stem cells of
mice was further in the green AgNPs synthesized using black cumin extract than
chemical AgNPs. IC50 (inhibitory concentrations) values for seed germination,
root and shoot length for 6 plants in green AgNPs exposures were higher than the
chemical AgNPs. These results suggest that cytotoxicity and phytotoxicity of the
green synthesized AgNPs were significantly less than wet-chemistry synthesized
ones. This study indicated an economical, simple and efficient ecofriendly
technique using leaves of N. sativa for synthesis of AgNPs and confirmed that
green AgNPs are safer than chemically-synthesized AgNPs.

26
10].Biosynthesis Of Zirconia (ZrO2) Nanoparticles Using Plant Extract

Abstract:

Zirconia (ZrO2) nanoparticles have a wide range of applications in various fields.

The green synthesis of metal oxide nanoparticles provides an advantage over the
chemical synthesis approach. Researchers are working on synthesis methods of
plant extracts to produce cost-effective and eco-friendly nanoparticles. Plant tissues
contain various phytochemicals i.e. phenolic compounds, flavonoids, alkaloids,
saponins, and amino [Link] compounds act as reducing, capping and
stabilizing agents during the formation of ZrO2 nanoparticles. In this review, we
focus on the biosynthesis of ZrO2 nanoparticles using different parts of plant
extracts. This review article has summarized research papers from 2018-2023
which describe the process of ZrO2 nanoparticles biosynthesis, characterization
and their application in different fields.

27
 MATERIALS AND METHODS

Preparation of seed extract

Black cumin seeds (Nigella sativa) were procured from a local supermarket in the
vicinity of Poonamallee. The seeds were ground into a fine powder using a
laboratory-grade grinder. Subsequently, 1 gram of the powdered black cumin seeds
was accurately weighed and dissolved in 100 mL of distilled water to prepare an
aqueous extract. The resulting mixture was subjected to heating using a controlled
heating mantle set at 70 °C for a duration of 15 min. Following heating, the
mixture was carefully filtered through Whatman No. 1 filter paper to remove any
insoluble particles, yielding a clear filtrate. The 80 mL filtrate, constituting the
aqueous extract of black cumin seeds, was promptly cooled and stored under
refrigeration conditions to preserve its integrity for further experimental
investigations

Extracting zirconium nanoparticles from cloves and black cumin seeds involves
using these plant materials as reducing and stabilizing agents for green synthesis.
Here's a step-by-step outline of methods typically employed:

1. Preparation of Plant Extracts

Materials: Cloves, black cumin seeds, distilled water and glassware.

Wash the plant materials thoroughly to remove impurities. Dry the cloves or cumin
seeds and grind them into a fine powder. Boil the powder in distilled water (e.g., 10
g in 100 mL water) for 10–30 minutes to extract bioactive compounds. Filter the
solution using Whatman filter paper or a similar filter and collect the plant extract.

28
2. Preparation of Zirconium Salt Solution

Materials: Zirconium precursors such as zirconium oxychloride (ZrOCl₂·8H₂O) or

zirconium [Link] a specific amount of zirconium precursor in distilled
water to prepare a 0.01–0.1 M [Link] the solution until completely dissolved.

3. Synthesis of Zirconium Nanoparticles

Mix the plant extract with the zirconium precursor solution in a 1:1 to 1:4 ratio
under constant [Link] the mixture at 60–80°C for 1–2 hours to initiate the
reduction [Link] the color change (e.g., pale to dark, depending on
nanoparticle formation).

4. Separation of Nanoparticles

Centrifuge the reaction mixture at 10,000–15,000 rpm for 15–30 minutes. Collect
the pellet and wash it with distilled water or ethanol to remove unreacted materials.
Dry the nanoparticles in an oven at 60–100°C or under vacuum.

5. Characterization of Nanoparticles

Use techniques like: UV-Vis Spectroscopy: To confirm the formation of zirconium

nanoparticles. Scanning Electron Microscopy (SEM) or Transmission Electron
Microscopy (TEM): To analyze particle size and morphology. X-ray Diffraction
(XRD): To determine crystallinity. Fourier-Transform Infrared Spectroscopy

29
(FTIR): To confirm functional groups from plant extracts. This green synthesis
method is cost-effective, eco-friendly, and avoids toxic chemicals. Let me know if
you'd like details on any step!

conductivity meter

ph meter

30
 RESULTS AND DISCUSSIONS

OD VALUES

CONCENTRATION OF CUMIN OD@ 520 NM

0.025 0.63

0.005 0.34

0.0025 0.33

0.0005 0.36

0.00075 0.32

CONCENTRATION OF CLOVE OD@ 550 NM

0.025 1.00

0.005 0.99

0.0025 0.77

0.0005 0.98

0.00075 0.93

31
PH METRY

CONCENTRATION OF CUMIN P.H

0.025 1.95

0.005 4.28

0.0025 4.50

0.0005 1.76

0.00075 1.50

CONCENTRATION OF CLOVE P.H

0.025 1.90

0.005 1.57

0.0025 3.50

0.0005 3.10

0.00075 1.45

32
CONDUCTOMETRY

CONCENTRATION OF CUMIN CONDUCTANCE

0.025 1.5

0.005 7.8

0.0025 05.2

0.0005 1.5

0.00075 11.8

CONCENTRATION OF CLOVE CONDUCTANCE

0.025 4.2

0.005 8.8

0.0025 0.8

0.0005 1.0

0.00075 11.4

33
 CONCLUSION
Plant extracts and their therapeutic activities have been applied since the beginning
of humanity through the traditional knowledge of their health benefits, even if, at
the time, little knowledge existed to justify their use. Over time and with
technological advances, scientists sought to understand the main compounds that
offer plant extracts medicinal properties. Plant extracts and essential oils feature
various molecules with a broad spectrum of activities not limited to
pharmacological potential, but that can also be applied in the food industry,
cosmetics, agriculture, and other fields.

Furthermore, these plant derivatives can be employed in the production of

nanomaterials with a variety of applications. The synthesis using plant products has
been widely investigated in recent years, demonstrating a variety of morphologies,
sizes, and properties. As reported in the literature, green synthesis reduces the
toxicity of the nanomaterial and the production of by-products throughout the
entire process, and consumes less energy. However, an important point to be
considered in this synthesis methodology is the type of extraction as well as the
adopted preparation of plant extract and essential [Link] type of extraction
dramatically influences the type of bioactive compounds that will be isolated,
consequently affecting its pharmacological properties and ability to create a
suitable medium for the synthesis and stabilization of nanostructures. Therefore, it
is essential to consider the physicochemical characteristics of the molecules of
interest, such as solubility and stability at high temperatures. In this context,
identifying such substances through advanced characterization techniques is
essential.

34
 BIBLIOGRAPHY
1.Süntar I. Importance of ethnopharmacological studies in drug discovery:
Role of medicinal plants. Phytochem. Rev. 2020;19:1199–1209. doi:
10.1007/s11101-019-09629-9. [DOI] [Google Scholar]

[Link] A. Phytomedicinal Potential Characterization of Medical Plants

(Rumex nervosus and Dodonaea viscose) J. Biochem. Technol.
2020;11:113–121. [Google Scholar]

[Link] M. Ancient medicine between hope and fear: Medicament,

magic and poison in the Roman Empire. Eur. Rev. 1999;7:37–51. doi:
10.1017/S1062798700003720. [DOI] [Google Scholar]

[Link] S.A. Potential anti-influenza effective plants used in Turkish folk

medicine: A review. J. Ethnopharmacol. 2021;265:113319. doi:
10.1016/[Link].2020.113319. [DOI] [PMC free article] [PubMed] [Google
Scholar]

[Link] G.B. Medicinal Plants for the Prevention and Treatment of

Bacterial Infections. Curr. Pharm. Des. 2005;11:2405–2427. doi:
10.2174/1381612054367481. [DOI] [PubMed] [Google Scholar]

[Link]éphane F.F.Y., Jules B.K.J., Batiha G.E.-S., Ali I., Bruno L.N. Extraction
of Bioactive Compounds from Medicinal Plants and Herbs. Nat. Med.
Plants. 2022 doi: 10.5772/intechopen.98602. [DOI] [Google Scholar]

[Link] D.C., Gu X.J., Xiao P.G. Medicinal Plants: Chemistry, Biology and
Omics. Woodhead Publishing; Sawston, UK: 2015. [Google Scholar]

[Link] V., Zhang L., Yan J., Fu Z., Yang T., Zeng H. Biochemistry of
Terpenes and Recent Advances in Plant Protection. Int. J. Mol. Sci.
2021;22:5710. doi: 10.3390/ijms22115710. [DOI] [PMC free article]

35
 [Link] I., Aboulmouhajir A., Kouisni L., Bekkaoui F., Yasri A. Plants
extracts and secondary metabolites, their extraction methods and use in
agriculture for controlling crop stresses and improving productivity: A
review. Acad. J. Med. Plants. 2018;6:223–240. doi:
10.15413/ajmp.2018.0139. [DOI] [Google Scholar]

[Link] L., Jha N., Meeran M., Kesari K., Beiram R., Ojha S.
Neuroprotective Potential of Limonene and Limonene Containing Natural
Products. Molecules. 2021;26:4535. doi: 10.3390/molecules26154535.
[DOI] [PMC free article] [PubMed] [Google Scholar]

[Link] A., Beserra F., Souza M., Totti B., Rozza A. Limonene: Aroma of
innovation in health and disease. Chem. Biol. Interact. 2018;283:97–106.
doi: 10.1016/[Link].2018.02.007. [DOI] [PubMed] [Google Scholar]

[Link] O., Bonfill M., Moyano E., Onrubia M., Mirjalili M.H., Cusido
R.M., Palazon J. Biotechnological Production of Taxol and Related Taxoids:
Current State and Prospects. Anti-Cancer Agents Med. Chem.
2012;9:109–121. doi: 10.2174/187152009787047761. [DOI] [PubMed]
[Google Scholar]

[Link] G.P., Vermaak I., Viljoen A.M., Lawrence B.M. Menthol: A

simple monoterpene with remarkable biological properties. Phytochemistry.
2013;96:15–25. doi: 10.1016/[Link].2013.08.005. [DOI] [PubMed]
[Google Scholar]

[Link] J. Alkaloids—Their Importance in Nature and Human Life.

Intechopen; London, UK: 2019. [(accessed on 17 January 2023)]. Available
online: [Link] [Google Scholar]

36
 [Link] S., Kondhare K., Giri A. Diversity in Chemical Structures
and Biological Properties of Plant Alkaloids. Molecules. 2021;26:3374. doi:
10.3390/molecules26113374. [DOI] [PMC free article] [PubMed] [Google
Scholar]

[Link] M.C., Pinto D.C.G.A., Silva A.M.S. Plant Flavonoids: Chemical

Characteristics and Biological Activity. Molecules. 2021;26:5377. doi:
10.3390/molecules26175377. [DOI] [PMC free article] [PubMed] [Google
Scholar]

[Link] K., Brodowska K.M. Natural Flavonoids: Classification,

Potential Role, and Application of Flavonoid Analogues. Eur. J. Biol. Res.
2017;7:108–123. [Google Scholar]

[Link] A.E.-M.M.R., El-Beltagi H.S., El-Salam S.M.A., Omran A.A.

Biochemical changes in phenols, flavonoids, tannins, vitamin E, β–carotene
and antioxidant activity during soaking of three white sorghum varieties.
Asian Pac. J. Trop. Biomed. 2012;2:203–209. doi:
10.1016/S2221-1691(12)60042-2. [DOI] [PMC free article] [PubMed]
[Google Scholar]

[Link] J., Puupponen-Pimiä R., Dauer A., Aura A.-M., Saura-Calixto F.

Tannins: Current knowledge of food sources, intake, bioavailability and
biological effects. Mol. Nutr. Food Res. 2009;53:S310–S329. doi:
10.1002/mnfr.200900039. [DOI] [PubMed] [Google Scholar]

[Link] R.G.L., Joaquim R.H.V.T., Sampaio S.F. Plantas Medicinais: O

Conhecimento e Uso Popular. Rev. APS—Atenção Primária Saúde.
2015;18:1–13. [Google Scholar]

37