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Chemistry DOI: 10.4172/2380-2391.1000106

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ISSN: 2380-2391

Research Article Open Access

Analytical and Biological Evaluation of Two Schiff’s Bases:

Spectrophotometeric Analysis of Copper (II) in Water and Soil Samples
Bhajanthri Natesh Kumar1, Suvardhan Kanchi2, Krishna Bisetty2 and Nimmagadda Venkata Vijaya Jyothi1*
1
Department of Chemistry, Sri Venkateswara University, India
2
Department of Chemistry, Durban University of Technology, South Africa

Abstract
Two novel ligands, (E)-N1-(2-hydroxy-5-nitrobenzlidene) isonicotinoylhydrazone (HNBISNH)/2-(4-fluoro
benzlideneamino)benzenethiol (FBBT) were synthesized and used for the quantification of copper (II) in various water
and soil samples. HNBISNH/FBBT interacts with copper (II) to form 1:1 [L:M] orange /brick red color complexes in
presence of phosphate buffer of pH 3.7/4.5 which increases the sensitivity of the complexes. Thus formed colored
complex followed the Beer’s law up to 1.6 and 1.9 mg L‑1 with an optimum concentration range over 0.11-1.3 mg L-1
and 0.10-1.7 mg L-1, the slope of Ringbom’s plot are found to be 0.3500 and 0.3300 for copper(II)-HNBISNH and
copper(II)-FBBT complexes, respectively. The molar absorptivity and Sandell’s sensitivity of copper (II)-HNBISNH and
copper(II)-FBBT complexes were calculated and found to be 4.91×104 L mol−1 cm−1, 6.1×104 L mol−1 and 0.0010 μg
cm−2 and 0.0014 μg cm−2 respectively. The detection limits was calculated and found to be 0.043 µg L−1 and 0.036 µg
L−1 with HNBISNH and FBBT respectively. Various optimum conditions such as effect of pH, reagent concentration,
accuracy, precision and reproducibility were investigated to improve the sensitive for the present method. The detailed
study of various excipients confirmed the high selectivity of the method. The proposed method was applied to the
determination of copper (II) in different water & soil samples obtained results were in good agreement with atomic
absorption spectrometric method (AAS).

Keywords: (E)-N1-(2-hydroxy-5-nitrobenzlidene) isonicotinoyl The aim of the present work was to provide a facile, sensitive and
hydrazone (HNBISNH); 2-(4-fluoro benzlideneamino) benzenethiol rapid non-extractive spectrophotometric method for the determination
(FBBT); Copper (II); Spectrophotometry; Biological activity; of trace amounts of copper (II) in different sample of environmental
Environmental samples importance. In this study, new analytical reagents (E)-N1-(2-hydroxy-
5-nitrobenzlidene) isonicotinoyl hydrazone (HNBISNH) and
Introduction 2-(4-fluorobenzlidene amino) benzenethiol (FBBT) was successfully
Copper is an essential trace nutrient to all high plants and synthesized for the determination of copper (II) in environmental
animals. In animals, including human it is found in primarily in the samples.
bloodstream, and in copper-based pigments. However, insufficient Experimental
amounts; copper can be poisonous and even fatal to organisms.
Copper also has a significant presence as a decorative metal art. Instrumentation
It can also be used as an anti-germ surface that can add to the anti- A HITACHI model U 3400 UV VIS NIR Spectrophotometer with
bacterial and antimicrobial feature of buildings such as hospitals [1]. 10 mm stopped glass cells was used. An ELICO model Li-129 was used
Copper has a high electrical and thermal conductivity, among pure for all pH measurements.
metals at room temperature [2]. On the other hand, toxic rule of
the metal ion is well recognized [3]. Heterocyclic azodyes have been Reagents
used as chromogenes in spectrophotometric determination of copper All reagents used were of analytical reagent grade and solutions were
(II) ions. These azodyes are 2-amino-5-bromopyridylazo resorcinol prepared with deionized distilled water unless mentioned. A phosphate
[4], 1-[pyridyl 2-azo]-naphthol (PAN) [5,6], and its derivatives are buffer solution prepared by adding: 50 mL of 0.25M disodium hydrogen
1-(2-thiazolylazo)-2-naphthol [7], 1-(5 bromo-2-pyridylazo)-2- orthophosphate (SD fine Chemicals, India) dissolved in 20 mL of 0.1M
naphthol-6-sulphonic acid [8], 2,6-bis(4-sulfo-1-hydroxy-2 naphth phosphoric acid (SD fine Chemicals, India) and pH was adjusted to
alazo) pyridine [9], 4-(1H-pyrazolo 3,4-d}pyrimidin-4-ylazo) benzene- 4.0/4.7 with 0.25 M disodium hydrogen orthophosphate solution and
1,3-diol [10], 4-(p-Nitrophenylazo)-2-amino-3-pyridinol [11], l-(2- diluted to 100 mL in a volumetric flask.
quinolylazo)-2,4,5 trihydroxybenzene [12], bromosulphonazo III [13]
and alizarin red S [14]. These dyes are highly sensitive but lack of the
selectivity. The coordination chemistry of hydrazones is an intensive
*Corresponding author: Nimmagadda Venkata Vijaya Jyothi, Department of
area of study of numerous transition metals. Hydrazones have been
Chemistry, Sri Venkateswara University, Tirupati-517 502, A.P, India, Tel: 91-
studied as a group of most useful spectrophotometric reagents. The 9912366219; E-mail: [email protected]
short comings of hydrazones were the lack of selectivity for metal
Received January 27, 2014; Accepted March 10, 2014; Published March 12,
ions, therefore much work have been devoted to the development 2014
of masking agents for use of hydrazones. Among these some of the
Citation: Natesh Kumar B, Kanchi S, Bisetty K, Vijaya Jyothi NV (2014) Analytical
hydrazone are 2,2’-dipyridyl-2-pyridylhydrazone [15] and 2-pyridine and Biological Evaluation of Two Schiff’s Bases: Spectrophotometeric Analysis of
carboxaldehyde isonicotinyl hydrazone [16]. Many techniques such Copper (II) in Water and Soil Samples. J Environ Anal Chem 1: 106.
as Atomic Absorption Spectrophotometry [17], Inductively Coupled doi:10.4172/10.4172/2380-2391.1000106

Plasma Atomic Emission Spectroscopy [18], Voltammetry [19] and Copyright: © 2014 Natesh Kumar B, et al. This is an open-access article
Spectrophotometry [20,21] have been reported for the determination distributed under the terms of the Creative Commons Attribution License, which
permits unrestricted use, distribution, and reproduction in any medium, provided
of copper (II). the original author and source are credited.

J Environ Anal Chem Volume 1 • Issue 1 • 1000106

ISSN: JREAC, an open access journal
Citation: Natesh Kumar B, Kanchi S, Bisetty K, Vijaya Jyothi NV (2014) Analytical and Biological Evaluation of Two Schiff’s Bases: Spectrophotometeric
Analysis of Copper (II) in Water and Soil Samples. J Environ Anal Chem 1: 106. doi:10.4172/2380-2391.1000106

Page 2 of 7

Synthesis of ligands Staphylococcus aureus) and gram negative organisms (Escherichia coli
and Pseudomonas aeruginosa) using disc diffusion method [22]. The
Preparation of (E)-N -(2-hydroxy-5-nitrobenzlidene)
1
Muller-Hinton agar was rigorously tested for the composition and pH.
isonicotinoyl hydrazone (HNBISNH): Accurately weighed 1.2g of
Further, the depth of the agar in the plate was a factor to be considered
2-hydroxy-5-nitrobenzaldehyde and 0.822g of isonicotinoyl hydrazide
in the disc diffusion method. The test plates were prepared by 0.5 mL
were transferred into a 50 mL round bottom flask. Ethyl alcohol was
of standard inoculums pipetted into a sterile petri plate; 20 mL of
used as solvent and the condensation process was carried out about
melted agar medium was then added to 10 cm petri plate and mixed
2 h at 60°C.The yellow colored product of HNBISNH was formed
well by gently swirling on the table top. The seeded plates are allowed
and further subjected to evaporate the solvent and collected the pure
to solidify and 10 µg mL-1 of complex compounds was impregnated on
solid compound with yield about 1.98g (86%). The melting point of
sterilized filter paper disc was carefully transferred in the agar plates.
HNBISNH was found to be in the range of 214-216°C with solubility
The chemical diffuses from the disc into the agar and place the chemical
in DMF. The preparation pathway of color forming ligand was shown
only around the disc. The solubility of the chemical and its molecular
in Scheme 1a.
size was determined using the chemical infiltration size around the
Preparation of 2-(4-fluorobenzlideneamino) benzenethiol disc.Standard disc of Gentamycin 10 µg (antibacterial agent) was
(FBBT): The physical state of 2-aminobenethiol was semi solid while served as positive controls for antimicrobial activity on the agar plates.
4-fluorobenzaldehyde was colorless oily liquid. Both reactants density The plates were incubated at 37°C for 24 h around the disc opaque area
values were taken into consideration for the synthesis of product. indicates that the compound, which diffused into the agar from the
Since those two reactants are nearly liquids accurately measured 0.48 disc, which inhibits the growth of the organism. The inhibition Zone
mL of 2-aminobenethiol and 0.48 ml of 4-fluorobenzaldehyde were (IZ) was recorded with the help of scale, and compared with control
transferred into a 50 mL round bottomed flask and methanol was containing standard antibiotic Gentamycindisc. The experiment was
used as a solvent. Reflux the reaction mixture for about 1h at 60°C. triplicated and the mean values are presented along with the standard
Pale yellow colored oily product was formed and further subjected deviation. Minimum Inhibitory Concentrations were determined by
rotor to expel the excess of solvent and obtained the pure product. This the Vollekova and Usman two fold dilution method [23,24].
product on long standing in ambient temperature converted to yellow
Sample preparation
solid. The melting point was in the range of 202-204°C with solubility
in hexane. The reaction procedure of color forming ligand was shown Procedure for water samples: One liter of the water sample
in Scheme 1b. collected from various sites outside the Tirupati city were acidified with
hydrochloric acid and filtered with a 0.45 µm filter. The samples were
Preparation of standard solutions pre-heated and evaporated to 250 mL. The copper (II) contents were
2.86 g of HNBISNH and 2.31 g of FBBT was dissolved in deionized determined according to aforesaid procedures. The obtained results
distilled water and diluted to 100 mL to get 0.1 M of stock solution. were compared with standard atomic absorption method in terms of
Working solution of HNBISNH and FBBT were prepared by student’s ‘t’-test and variance ratio ‘f”-test.
appropriate dilution and the solutions were stable for several months. A
Procedure for soil samples: Soil samples collected from industrial
stock solution of copper (II) (1.0 mg mL-1) was prepared by dissolution
areas surrounding Tirupati city were accurately weighed (2.0 g) in a 100
of 0.250 g copper sulphate pentahydrate (SD fine Chemicals, India) in
mL beaker. The content was digested as per to the reported method,
100 mL standard flask and further dilutions as required were employed.
[25] then 10 mL concentrated HNO3 and 2.0 mL of 70% HClO4 (v/v)
General procedure was added and heated for 1 h. The mixture was filtered through a
Whatman No. 40 filter paper into a 250 mL calibrated flask and its pH
In the first step an aliquot containing not more than 1.0–100 µg was adjusted to desired value and diluted to mark with deionised water.
mL-1of copper (II) was transferred into 25 mL calibrated flask, 3.0 mL In all of real and synthetic amount of copper (II) ion was found by
of 0.005 M HNBISNH and 5 mL of phosphate solution with pH 4.0 standard addition method.
solution were added successively and the mixture was diluted up to
the markwith deionized distilled water. The first step was repeated Results and Discussion
for second ligand, FBBT as well. The appearance of the orange and
brick red color chromophores was instantaneous and absorbance Absorption spectra
was measured at 480 nm and 520 nm against reagent blank solution. Under the optimal experimental condition, the absorption spectra
The reagent blank was prepared in a similar manner without copper of HNBISNH/FBBT and copper (II)–HNBISNH/FBBT complexes
(II). All measurements were carried out at ambient temperature. The were scanned. The absorption maximum of reagent blanks HNBISNH/
copper(II) content in an unknown sample was determined using a FBBT, were measured at 370 nm and 395 nm, whereas copper (II)–
calibration graph. HNBISNH/FBBT complexes gave an absorption peak at 460 nm and
Procedure for evaluation of biological activity of complexes 530 nm. The difference (bathochromic shift) of the two peaks was 90 nm
and 145 nm, and could be obviously distinguished. Thus, the absorption
To a methanolic solution of copper (II) sulphate, a hot methanolic peaks at 460 nm and 530 nm were chosen as the determination of wave
Solution of the HNBISNH/FBBT ligand was added slowly with stirring. length for copper (II)–HNBISNH/FBBT complexes as illustrated in
The mixture was refluxed on a hot water bath. It was concentrated Figure 1.
under reduced pressure to two-third the original volume and cooled.
The solid that separated out was filtered, washed with water, hot Effect of pH
methanol and ether and was vacuum dried to obtain solid complexes. pH studies were carried out on the peak height of copper (II)-
Antibacterial activity of the two synthesized complex compounds HNBISNH/FBBT complexes at various concentrations (0.001–0.10M)
was determined with gram positive organisms (Bacillus subtillis and by fixing 0.005M HNBISNH and FBBT concentration. The pH of

J Environ Anal Chem Volume 1 • Issue 1 • 1000106

ISSN: JREAC, an open access journal
Citation: Natesh Kumar B, Kanchi S, Bisetty K, Vijaya Jyothi NV (2014) Analytical and Biological Evaluation of Two Schiff’s Bases: Spectrophotometeric
Analysis of Copper (II) in Water and Soil Samples. J Environ Anal Chem 1: 106. doi:10.4172/2380-2391.1000106

Page 3 of 7

phosphate buffer was changed over a range of 2.0-7.0 and the peak
height were measured for each concentration level of copper (II). At 0.4
all concentration levels of copper (II)-HNBISNH/FBBT complexes, Cu(II) HNBISNH Cu(II) FBBT
0.35
maximum absorbance were found between pH 3.0 and 5.0. Therefore,
0.3
a pH 3.7 for HNBISNH and 4.5 for FBBT of phosphate buffer system
0.25

Absorbance
was chosen throughout the study as shown in Figure 2.
0.2
Effect of ligand concentration on absorbance 0.15

The effect of concentrations of both ligands (HNBISNH and 0.1

FBBT), on the peak height was investigated at pH 3.7 and 4.5 in a 0.05

phosphate buffer system. The concentrations of HNBISNH and FBBT 0

was varied over the range 0.001– 0.10 M. Maximumpeak height was

0. 1
5
01

0. 2
5

0. 3
5

0. 4
5
05

5
06
65

0. 7
5
08

5
09

5
1
00
00

01
0
02

0
03

0
04

05

00
07

08

09
0.
0.

0.

0.

0.

0.

0.
0.

0.

0.
0.

0.

0.

0.

0.

0.
obtained at a concentration of 0.04 M for HNBISNH and 0.01 M FBBT Ligand Concentration (Moles/Litre)
as color developing reagents for lower concentration level of copper Figure 3: Effect of ligand concentration on the absorption of copper (II)
(II) in the sample solution. Further, increase in HNBISNH and FBBT complexed with HNBISNH and FBBT (Optimum conditions:- pH:3.7/4.5,
concentration, slight decrease in the peak height was observed. Hence, Temperature: 32 ± 5°C).
0.04 M for HNBISNH and 0.01 M FBBT was optimized for all further
studies as represented in Figure 3. (32 ± 5°C) and remained stable for 30 h and 60 h for copper (II)–
HNBISNH/FBBT complexes, respectively.
Effect of time and temperature on absorbance
The reaction was instantaneous for the both complexes, but these
Stability of the chromophoric system
systems attained maximum and constant absorbance just after the After mixing all the components, the absorbance related to
dilution to volume (25 mL in volumetric flask) at room temperature both complexes reached its maximum in less than 1 min at ambient
temperature and remain stable for 30 h [copper (II)–HNBISNH]
and 60h [copper (II)-FBBT] in aqueous solution. Due to stability of
0.8 the complex and clarity of complex solution, the extraction step was
HNBISNH FBBT Cu (II)-HNBISNH Cu (II)-FBBT omitted to reduce usage of environmental hazardous solvents, which
0.7
made the method eco-friendly.
0.6
Composition of the complexes
0.5
The stoichiometry of the complexes was identified using the molar
Absorbance

0.4 ratio at the optimum conditions discussed above. Figures 4a and 4b

show that maximum absorption was found to be at molar ratio 1.0
0.3
by varying the concentrations of HNBISNH and FBBT, representing
0.2 the interaction of one ligand molecule with metal in the complex
formation. Accordingly, the results indicated that the stoichiometric
0.1
ratio was (1:1) [M:L]. The composition of the both complexes [copper
0 (II)-HNBISNH/FBBT] were determined using Job’s continuous
0 100 200 300 400 500 600 700 800
Wave length (nm)
variation method indicates the formation of 1:1 (M:L) complexes.
Ringbom plot’s for copper (II)-HNBISNH/FBBT complexes
Figure 1: Absorption spectra for copper (II)-HNBISNH/FBBT complexes
against reagent blank (Optimum conditions:- pH:3.7/4.5, Ligand Ringbom’s plot is the standard protocol adopted to know the
concentration:0.04/0.01M, Temperature: 32 ± 5°C). optimum range of concentration for chromophoric system that obeys
Beer’s law. The plots were drawn between log C of copper (II) and (1-
T) (where T is the transmittance). The plots was sigmoid shape with
a linear segment at intermediate absorbance values 0.1–1.0, 0.2-1.6
0.45 and concentration values 1.20-3.1 μg mL-1 and 1.22-3.4 μg mL-1 for
0.4 copper (II)-HNBISNH and copper (II)-FBBT respectively. The slope
0.35
of Ringbom’s plot from Figure 5 are found to be 0.3500 for copper
Absorbance

0.3
0.25
(II)-HNBISNH and 0.3300 for copper (II)-FBBT complexes. Hence,
0.2
the ratio between the relative error in concentration and photometric
0.15
Cu(II) HNBISNH Cu(II) FBBT errors are 2.50, 2.20 for found to be concentration of 0.0250, 0.0220
0.1 having 1.0% photometric error.
0.05
0
Sensitivity and calibration curves
2 2.5 3 3.5 3.7 4 4.5 4.7 5 5.5 6 6.5 7
The complexes obey Beer’s law up to 1.7 and 2.0 mg L‑1 with an
pH
optimum concentration range between 0.11 - 1.4 mg L-1 and 0.10 -
Figure 2: Effect of pH on the absorption of copper (II) complexed with 1.8 mg L-1 for copper (II)-HNBISNH/FBBT systems respectively. The
HNBISNH and FBBT (Optimum conditions:- Ligand concentration:0.04/0.01M, molar absorptivity of complexes at 460 and 530 nm and at pH 3.7 and
Temperature: 32 ± 5°C).
4.5 was calculated 4.91×104 L mol−1 cm−1 and 6.1×104 L mol−1 cm−1

J Environ Anal Chem Volume 1 • Issue 1 • 1000106

ISSN: JREAC, an open access journal
Citation: Natesh Kumar B, Kanchi S, Bisetty K, Vijaya Jyothi NV (2014) Analytical and Biological Evaluation of Two Schiff’s Bases: Spectrophotometeric
Analysis of Copper (II) in Water and Soil Samples. J Environ Anal Chem 1: 106. doi:10.4172/2380-2391.1000106

Page 4 of 7

respectively. Sandell’s sensitivity for both complexes were found to be

0.0010 μg cm−2 and 0.0014 μg cm−2 for copper (II)-HNBISNH/FBBT 1.8
systems respectively. The correlation coefficient (γ) for the complexes Cu(II) HNBIS NH Cu(II) FBBT
1.3
of copper (II)-HNBISNH/FBBT experimental data was 0.9996 and
0.99998 respectively. The composition of the both complexes [copper 1.4

(II)-HNBISNH/FBBT] were determined using Job’s continuous 1.2

variation method indicates the formation of 1:1 (M:L) complexes. The
1
stability constants corresponding to copper (II)-HNBISNH/FBBT

(1-T)
was determined and found to be 5.9×1015 and 6. 7×1015 (1:1, M:L) 0.8
respectively. The calibration graphs were obtained at the optimum 0.6
working conditions as shown in Figures 6a and 6b.
0.4
Effect of excipients 0.2

To understand the selectivity of the present method using two 0

ligands (HNBISNH/FBBT), the effect of potentially interfering species 0 0.5 1 1.5 2 2.5 3 3.5 4
Log. C
on the copper (II) determination was studied and carried out by adding
a known concentration of excipients to copper (II) solution of 0.2 Figure 5: Ringbom plot’s for copper (II) complexed with HNBISNH and
µg mL-1. Several anions and captions were studied in detail. Table 2 FBBT (Optimum conditions:- pH:3.7/4.5, Ligand concentration: 0.04/0.01M,
Temperature: 32 ± 5°C).
summarizes the tolerance limits of interfering ions in the determination
of 0.2 µg mL-1 copper (II). The tolerance limit was taken as the amount
causing an error of ± 2% at the peak height.
160
Method evaluation 140
The proposed method was critically evaluated with regard to 120
reproducibility, accuracy, and detection limit for analysis of copper (II) 100
Area/m2

in various environmental samples.

80

60
Plot Area
1.6 40
1.4 20
1.2 0
1 0 0.005 0.01 0.015 0.02 0.025 0.03
Absorbance

Cu(II), µg/L
0.8
0.6 Figure 6a: Calibration plots for the determination of copper (II) with HNBISNH.

0.4
0.2
0
0 1 2 3 4 5
Molar Ratio

Figure 4a: Molar ratio for copper (II)-HNBISNH complex.

2
1.8
1.6
1.4
Absorbance

1.2
1
0.8 Figure 6b: Calibration plots for the determination of copper (II) with FBBT.
0.6
0.4 Reproducibility
0.2
0 To test the reproducibility of the present method, four repetitive
0 1 2 3 4 5 analyses of each sample was studied. The RSD (%) values for copper
Molar Ratio (II)-HNBISNH/FBBT were found to be 0.600 and 0.598 (Table 1). A
% SD values obtained from the present method ranges 0.42–1.95 as
Figure 4b: Molar ratio for copper (II)-FBBT complex.
summarized in Tables 3 and 4.

J Environ Anal Chem Volume 1 • Issue 1 • 1000106

ISSN: JREAC, an open access journal
Citation: Natesh Kumar B, Kanchi S, Bisetty K, Vijaya Jyothi NV (2014) Analytical and Biological Evaluation of Two Schiff’s Bases: Spectrophotometeric
Analysis of Copper (II) in Water and Soil Samples. J Environ Anal Chem 1: 106. doi:10.4172/2380-2391.1000106

Page 5 of 7

HNBISNH FBBT Samples Present Method ASS Method

Optical Characteristics
Values Values Cu (II) [µg L-1]
Color Orange Brick Red Added Found Recovery ± t-Teste F-Testf Found Recovery ±
λmax[nm] 460 530 SDd SDd
Stability 30 h 60 h Water samples
Beer’s law range [mg L-1] 0.11 - 1.4 0.10 - 1.8 SWS 2.0 1.96 9.40 ± 0.66 7.00 2.69 1.97 99.00 ± 0.99
Molar absorptivity [L mol-1 cm-1] 4.91×104 6.1×104 NWS - 0.23 - - - 0.24 -
Sandell_s sensitivity [µg cm ] -2
0.0010 0.0014 SWS 3.0 2.94 99.30 ± 0.32 2.66 0.731 2.96 99.33 ± 0.84
Regression equation (Yb) NWS - 0.20 - - - 0.22 -
Slopea 0.1715 0.1728 SWS 4.0 3.97 99.25 ± 0.68 1.90 1.02 3.98 99.50 ± 0.58
Interceptb 0.0163 0.0152 NWS - 0.20 - - - 0.21 -
Correlation coefficient [r] 0.9994 0.9998 SWS 5.0 4.98 99.60 ± 0.80 3.85 0.15 4.99 99.80 ± 0.42
Relative standard deviation [%]c 0.600 0.598 Soil samples
Range of error (95% confidence level) ± 1.186 ± 1.044 SSS 2.0 1.94 98.10 ± 1.65 2.96 0.40 1.95 98.50 ± 1.30
Detection limit [µg L ]
-1
0.043 0.036 NSS - 0.31 - - - 0.31 -
% Error 0.101 0.108 SSS 3.0 2.95 98.23 ± 0.70 3.00 1.16 2.95 98.73 ± 0.64
NSS - 0.24 - - - 0.25 -
a
Experiments performed under optimized conditions (see text),
b
Y = ax + b, where x is the concentration of analyte in µg ml-1, SSS 4.0 3.95 99.10 ± 1.46 2.22 0.89 3.96 99.45 ± 1.98
c
n=4 NSS - 0.30 - - - 0.30 -
Table 1: Optical characteristics for the analysis of copper (II) with HNBIFBBT using SSS 5.0 4.99 99.45 ± 0.77 1.88 1.05 4.98 99.40 ± 0.90
spectrophotometer. a
Samples collected around Amaraja Batteries,
b
Indsutrial Estate, Renigunta,
c
Chandragiri,
Tolerance limit/g L-1 Interfering ions
d
n=4,
e
P≥0.000284,
>50000 K(I),Na(II), Ca(II),Mg(II), Al(III), Cr(III)Cl-,Br-,PO33-, CO33,SO42- f
P= 0.05
>10000 Zn(II), Hg(II), NO3
Table 4: Analytical data for the determination of copper(II) with FBBT in various
>5000 Pb(II), Cd(II) water and soil samples.
>800 Co(II),Ni(II), Fe(II),Fe(III), Mn(II)
a
Can be masked up to 10000g L-1by the addition of 2 mL of EDTA, thiosulfate, Accuracy
cyanide.
b
Can be masked up to 5000 g L-1 by the addition of 2 ml of 2 % citrate solution. The accuracy of the proposed method was evaluated by comparing
c
Can be masked upto 800 g L-1 by the addition of 1 % thiocyanate the results with those obtained by the AAS method. The results showed
Table 2: Tolerance limits of interfering ions on the determination of 0.2 µg mL- in Tables 3 and 4 reveals that the good correlation between the two
1
copper (II) ion. methods indicative of present method was almost sensitive to that of
AAS method.
Present Method ASS Method
Detection limit
Cu (II) [µg L-1]
Samples
Recovery ± Recovery ± Under optimized conditions the detection limit for determination
Added Found t-Teste F-Testf Found
SDd SDd
of copper (II) by using present method (signal to noise ratio = 2) was
Water samples
0.043 µg L−1 and 0.036 µg L−1 with HNBISNH and FBBT respectively.
SWS 2.0 1.98 97.50 ± 0.74 6.40 2.30 1.97 98.50 ± 0.72
NWS - 0.24 - - - 0.24 -
Analytical applications
SWS 3.0 2.94 98.46 ± 0.44 2.38 0.74 2.96 98.66 ± 0.50
NWS - 0.21 - - - 0.22 - The present method was applied to the determination of copper
SWS 4.0 3.92 98.15 ± 0.42 1.82 0.96 3.95 98.75 ± 0.60 (II) in different water and soil samples using new synthesis ligands,
NWS - 0.17 - - - 0.18 -
HNBISNH/FBBT. This method was comparable with the standard
SWS 5.0 4.96 99.55 ± 0.60 3.50 0.94 4.98 99.60 ± 0.55
AAS method in terms of students‘t’ test and ‘f’ test as shown in Tables
Soil samples
3 and 4. The analytical data summarized in Tables 3 and 4 suggest that
SSS 2.0 1.91 95.00 ± 1.26 3.17 0.64 1.92 95.50 ± 1.28
the percentage of copper (II) recovery from water and soil samples
NSS - 0.29 - - - 0.29 -
SSS 3.0 2.88 96.22 ± 0.46 4.34 1.32 2.90 97.10 ± 0.64
ranges from 96.00 to 99.80% which is more reliable and sensitive
NSS - 0.10 - - - 0.10 - than the other methods. The recovery percentage of the copper (II)
SSS 4.0 3.90 97.50 ± 1.66 2.66 0.87 3.92 98.00 ± 0.98 represented in the Tables 3 and 4 indicates that their order in various
NSS - 0.32 - - - 0.32 - environmental systems under study are as follows:
SSS 5.0 4.94 98.80 ± 0.54 1.98 0.40 4.95 99.00 ± 0.89
Water samples with FBBT > Water samples with HNBISNH
a
Samples collected around Amaraja Batteries
b
Indsutrial Estate, Renigunta
c
Chandragiri, Soil samples with FBBT> Soil samples with HNBISNH
d
n=4
e
P≥0.000284 It is evident from the previous data that the proposed method
f
P= 0.059
was simple, rapid and sensitive for the determination of copper
Table 3: Analytical data for the determination of copper (II) with HNBISNH in (II) in different samples of environmental importance. The results
various water and soil samples.

J Environ Anal Chem Volume 1 • Issue 1 • 1000106

ISSN: JREAC, an open access journal
Citation: Natesh Kumar B, Kanchi S, Bisetty K, Vijaya Jyothi NV (2014) Analytical and Biological Evaluation of Two Schiff’s Bases: Spectrophotometeric
Analysis of Copper (II) in Water and Soil Samples. J Environ Anal Chem 1: 106. doi:10.4172/2380-2391.1000106

Page 6 of 7

showed that the calculated values (Tables 3 and 4) did not exceed the Conclusion
theoretical values. Therefore, there is no significant difference between
In this present investigation, evaluation of two ligands, HNBISNH
the proposed and the standard method, indicating that the developed
and FBBT were successfully study the analytical performance
method is as accurate and precise as the standard AAS method.
(complex affinity) experimentally. Due to the high complexing
Biological activity of the complexes ability of the ligands, the developed analytical method/applications
for the determination of copper (II) become facile, sensitive, rapid
Antibacterial activity of copper (II)-HNBISNH/FBBT synthesized and selective. This non-extractive spectrophotomeric method is eco-
complexes were studied against the gram positive bacteria (Bacillus friendly and effectively applied to analyze copper (II) in different water
subtillis and Staphylococcus aureus) and gram negative bacteria and soil samples. Moreover, antibacterial activity copper (II)-FBBT
(Escherichia coli and Pseudomonas aeruginosa) at a concentration of synthesized complex was greater than that of copper (II)-HNBISNH.
10µg mL-1 by disc diffusion method. The antibacterial activity was more This antibacterial activity facilitates the biological importance of the
effective with copper (II)-FBBT synthesized complex with 23.2 mm synthesized ligands to control diseases effectively.
to 16.5 mm Inhibition Zone than copper (II)-HNBISNH synthesized Acknowledgements
complex with an inhibition zone of 17.5 to 14.2 mm; the antibacterial
BNK is grateful for the financial support provided by the University Grant
activity of both the complexes was more when compared to that of the Commission, New Delhi, India in the form of JRF-BSR fellowship to carry out
control Gentamycin – Standard with 9.33 mm to 12.26 mm Inhibition this work. One of the authors is highly thankful to the Department of Botany, S.V.
Zone. In this experiment, copper (II)-HNBISNH synthesized complex, University, India for biological evaluation of the compounds and complexes.

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ISSN: JREAC, an open access journal
Citation: Natesh Kumar B, Kanchi S, Bisetty K, Vijaya Jyothi NV (2014) Analytical and Biological Evaluation of Two Schiff’s Bases: Spectrophotometeric
Analysis of Copper (II) in Water and Soil Samples. J Environ Anal Chem 1: 106. doi:10.4172/2380-2391.1000106

Page 7 of 7

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