ISSN 1061-9348, Journal of Analytical Chemistry, 2009, Vol. 64, No. 11, pp. 1193–1198. © Pleiades Publishing, Ltd.

, 2009.

ARTICLES

Bromatometric Assay of Simvastatin

in Pharmaceuticals1
Kalsang Tharpa and Kanakapura Basavaiah
Department of Chemistry, University of Mysore Manasagangotri, Mysore-570006, India
e-mail: basavaiahk@yahoo.co.in
Received September 9, 2008; in final form March 3, 2009

Abstract—Titrimetric and spectrophotometric methods are proposed for the determination of simvastatin
(SMT) in bulk drug and in tablets. The methods employ bromate-bromide mixture in acid medium as the bro-
minating agent and iron (III) and thiocyanate as auxiliary regents. In titrimetry, SMT is treated with a measured
excess of bromate-bromide mixture in HCl medium, and after a definite time, the unreacted bromine is deter-
mined iodometrically. In spectrophotometric method, the residual bromine is reduced by iron (II) and the result-
ing iron (III) is complexed with thiocyanate, and the absorbance is measured at 470 nm. In both methods, the
amount of in situ generated bromine corresponds to the SMT content. The experimental conditions are opti-
mized. Titrimetry is applicable over 1–10 mg range and the calculations are based on the molar ratio of 1 : 0.666
(SMT : KBrO3). In spectrophotometric method, Beer’s law is obeyed over the concentration range 1–10 µg/mL.
The calculated molar absorptivity is 3.02 × 104 L/mol cm and the corresponding sandel sensitivity being
0.0081 µg/cm2. The limit of detection (LOD) and limit of quantification (LOQ) are calculated to be 0.10 and
0.31 µg/mL, respectively. The intra-day and inter-day precision calculated from the analysis of pure SMT were
less than 2 and 2.7%, respectively. The methods were satisfactorily applied to the determination of SMT in tab-
lets, and no interferences from common tablet excipients were observed. The validity of the methods was fur-
ther ascertained by parallel assay by an established technique and by recovery studies.
DOI: 10.1134/S1061934809110161

1 INTRODUCTION in research/quality control laboratories in many devel-

oping/underdeveloped countries.
Simvastatin (SMT), chemically known as (1S, 2S,
8S, 8aR)-1,2,6,8,8a-hexahydro-1-(2-((2R, 4R)-tera- From the pharmaceutical standpoint, it is highly
hyro-4-hydroxy-6-oxo–2H-pyran-2-yl)-2,6-dimethyl- desirable to employ accurate, precise, reasonably rapid
napthalen-8-yl)2,2-dimethylbutanoate (Figure), belongs and cost-effective methods for the assay of drugs. In
to the group of cholesterol-lowering lactones known as this respect, titrimetry and visible spectrophotometry
statins which, in 2007, have been identified as being still continue to be of value in pharmaceutical analysis.
among the most widely prescribed drugs in the world. There are two reports on the use of visible spectropho-
Statins lower cholesterol by inhibiting the synthesis of tometry for the determination of SMT. One report [16]
movalinic acid, which is the key precursor in choles- describes three methods. One procedure is based on the
terol synthesis. SMT has been shown to be effective as reduction of iron (III) by SMT to iron (II) and subse-
an antilipemic agent. It is administered as a pro drug, quent formation of Prussian blue with ferricyanide
and in the liver it is hydrolysed to the β-hydroxy acid
form [1]. The official method for the quantitative deter-
mination of SMT drug substance is liquid chromatog- OH O
raphy with UV detection at 238 nm [2]. Not many
methods are found in the literature for the determina- O
O
tion of SMT in pharmaceuticals. Methods are based on H
the use of such techniques as HPLC [3–8], HPTLC [9], H3C
cyclic voltammetry [10], miscellar electrokinetic chro- O H
H3C H
matography [11] and UV-spectrophotometry [8, 12– CH3 CH3
15]. However, many of these methods [3–12] require
expensive instrumental setup, costly reagents/solvents,
elaborate multi-step extraction/clean up procedures. H3C
Moreover, these techniques are not so easily available
1 The article is published in the original. Structure of Simvastatin.

1193
1194 KALSANG THARPA, KANAKAPURA BASAVAIAH
measurable at 730 nm. In the other two procedures, the
iron (II) formed is chelated with 1,10-phenanthroline or
2,2'-bipyridine followed by measurement of absor-
bance at 480 or 490 nm. Very recently a procedure
based on the formation of yellow colour on binding
SMT with ferric nitrate in concentrated nitric acid has
been reported by Vijaya et al. [17]. However, literature
survey revealed that no titrimetric methods have ever
been reported for the determination of SMT. During the
preliminary studies, SMT was found to react with
in situ generated bromine in HCl medium which
prompted the authors to develop titrimetric and spectro-
photometric methods for the determination of drug
based on this reaction. This paper describes the study
and evaluation of the variables which govern the inter-
action of SMT and in situ bromine in order to provide
the basis for the simple, sensitive and cost-effective tit-
rimetric and spectrophotometric methods for the deter-
mination of SMT. The methods use bromate-bromide
mixture in HCl medium as the brominating agent and
thiocyanate as the complexing agent. The proposed
methods have the advantage of accuracy and precision
besides being free from interference from common tab-
let excipients.
MATERIALS AND METHODS
Apparatus. A systronics model 106 digital spectro-
photometer provided with 1-cm matched quartz cells
was used for absorbance measurements.
Reagents. All chemicals used were of analytical
reagent grade and distilled water was used throughout
the study. A standard stock solution of bromate-bro-
mide equivalent to 2 mM KBrO3–20 mM KBr was pre-
pared by dissolving accurately weighed 0.3340 g of
KBrO3 and 2.38 g of KBr in water and diluting to vol-
ume in a one-litre volumetric flask, and used in titrime-
try. A stock standard solution of KBrO3−KBr equivalent
to 300 µg/mL KBrO3 was prepared by dissolving accu-
rately weighed 30 mg of KBrO3 and 300 mg of KBr in
water and diluting to mark in a 100 mL calibrated flask.
It was diluted with water to obtain a working concen-
tration of 30 µg/mL in KBrO3. Hydrochloric acid (2 and
5 M), sodium thiosulphate (0.012 M), potassium thio-
cyanate (3 M), KI (10%) and starch indicator (1%)
were prepared in the usual way. Ferrous ammonium
sulphate, FAS (600 µg/mL) was prepared by dissolving
0.15 g of the chemical in water in the presence of 2 mL
of dilute H2SO4 and diluted to volume with water in
250 mL calibrated flask. A stock standard solution of
drug containing 1 mg/mL SMT was prepared by dis-
solving accurately weighed pure SMT in 3 : 2 acetic
acid and diluting with the same acid, and used in titri-
metry. For spectrophotometric work, the stock solution
was diluted to 40 µg/mL with 3 : 2 acetic acid.
Tablets containing SMT were purchased from local
commercial sources for investigation. Twenty tablets
were weighed accurately and finely powdered. An
amount of the powder equivalent to 100 mg of SMT
JOURNAL OF ANALYTICAL CHEMISTRY
was weighed into a 100 mL calibrated flask, 60 mL of
3 : 2 acetic acid were added and the content was shaken
for 15–20 min, diluted to the mark with same acid,
mixed well and filtered using a Whatman no. 42 filter
paper. First 10 mL portion of the filtrate was discarded
and a convenient aliquot (say 5 mL) of the subsequent
portion of the filtrate was assayed by titrimetry. The
same tablet extract was appropriately diluted with 3 : 2
acetic acid before analysis by spectrophotometry.
Procedures. Titrimetry. A 10 mL aliquot of standard
drug solution containing 1–10 mg of SMT was accu-
rately measured and transferred into a 100 mL Erlenm-
eyer flask. The solution was acidified by adding 5 mL
of 2 M HCl and 1 mL of glacial acetic acid. 10 mL of
bromate-bromide mixture (2 mM in KBrO3) was added
to the flask by means of a pipette with swirling and the
flask was stoppered and kept aside for 2–3 min with
occasional swirling. Then, 5 mL of 10% KI solution
was added and the liberated iodine titrated with
0.012 M thiosulphate to a starch end point. A blank
titration was performed taking 10 mL of 3:2 acetic acid
in place of drug solution. The amount of SMT in the ali-
quot was calculated from:
VSMw
SMT ( mg ) = ------------------ ,
0.666
where V - KBrO3 reacted, mL, S - Strength of KBrO3 in
M and Mw - relative molecular mass of SMT.
Spectrophotometry. Different aliquots (0, 0.25, 0.5,
1.0, 1.5, 2.0, 2.5 mL) of standard 40 µg/mL SMT were
accurately transferred into a series of 10 mL calibrated
flasks. To each flask, 1mL of 5 M HCl was added fol-
lowed by 1 mL of bromate-bromide (30 µg/mL in
KBrO3). The flasks were stoppered, content was mixed
and kept aside for 10 min with occasional swirling.
Then, 1mL of FAS (600 µg/mL) was added to each
flask, and after 5 min, 1 mL of 3 M thiocyanate was
added and diluted to the mark with water. The absor-
bance of each solution was measured at 470 nm against
a water blank. A standard graph was prepared by plot-
ting the measured absorbance Vs concentration (µg/mL)
of SMT. The concentration of the unknown was read
from the calibration graph or computed from the
regression equation derived using the Beerís law data.
RESULTS AND DISCUSSION
Potassium bromate in the presence of a large excess
of bromide in acid medium is valuable oxidimetric/bro-
minating agent and has been successfully employed in
the assay of many drug substances [18–23]. The present
communication deals with the titrimetric and spectro-
photometric assay of SMT using in situ generated bro-
mine as a brominating agent. The proposed methods are
indirect and are based on the determination of residual
bromine after the reaction between the drug and bro-
mine is judged to be and rely on different reaction
schemes.
Vol. 64 No. 11 2009
 BROMATOMETRIC ASSAY OF SIMVASTATIN
Method development. Titrimetry. Direct titration
of SMT with bromate-bromide was not successful.
Assay by back titration was found feasible, and hence
several experimental variables were optimized to get
accurate and precise results. Accurate and reproducible
stoichiometry was obtained when 2 M HCl concentra-
tion was maintained in addition to acetic acid present in
the drug solution. Hence, 5 mL of 2 M HCl in a total
volume of 15 mL (0.66 M overall) were used in the
study. 1 mL of glacial acetic acid was found necessary
to clear little turbidity that was formed on adding HCl.
The bromination reaction was found to be complete in
2–3 min and it is essential to terminate the reaction at
the end of the 3rd min but not before the 2nd min after
adding bromate-bromide mixture. Beyond 3 min and
upto 60 min little more bromine was found to be con-
sumed but without yielding a definite reaction stoichi-
ometry. Hence, the reaction time was stricted to
2−3 min. For the range investigated (1–10 mg) the reac-
tion yielded a 3 : 2 stoichiometry (SMT : KBrO3). Out-
side this range and reaction time, non-stoichiometric
results were obtained.
Spectrophotometry. Complex formation reaction
involving iron (III) and thiocyanate has been the basis
for the sensitive determination of the former in a variety
of matrices [24]. This reaction in combination of bro-
mate-bromide mixture has been applied for the assay of
some pharmaceuticals [18–20]. The present method is
based on the bromination of SMT by a measured excess
of in situ generated bromine in acid medium, reduction
of residual bromine by fixed amount of iron (II) and
subsequent complexation of iron (III) formed with thio-
cyanate which is measured at 470 nm. When a fixed
concentration of in situ generated bromine reacts with
increasing concentration of SMT, there occurs a con-
comitant fall in the bromine concentration. When the
unreacted bromine is reduced by a fixed concentration
of iron (II), there will be a proportional decrease in the
concentration of iron (III). This is observed as a propor-
tional decrease in the absorbance of iron (III)-thiocyn-
ate complex with increase in SMT concentration,
which formed the basis for the assay of drug.
The conditions for the determination of iron (III)
with thiocyanate are well established [24]. Hence, var-
ious parameters associated with the bromination of
drug, and subsequent reduction of residual bromine by
iron (II) where optimized. 1 mL of 5 M HCl in a total
volume of about 5 mL for bromination and reduction
steps was used, and the same quantity of HCl in a total
of 10 mL was used for the complexation step.
Considering 8 µg/mL as the upper limit of iron (III)
that could be accurately determined by thiocyanate
method, 30 µg/mL KBrO3 in the presence of a large
excess of bromide was found to generate it from
56 µg/mL FAS under the specified conditions. How-
ever, a slight excess (60 µg/mL) FAS was used to ensure
a quantitative reaction. Hence, 1 mL each of 30 µg/mL
KBrO3 (with excess of KBr) and 600 µg/mL FAS were
JOURNAL OF ANALYTICAL CHEMISTRY Vol. 64 No. 11
1195
used in the study. The bromination step took 5 min
while the reduction and subsequent complexation steps
were instantaneous. Developed colour was stable for at
least 60 min in the presence of the reaction product.
Two blanks were prepared in this study. The reagent
blank which contained optimum concentrations of all
reactants except SMT gave maximum absorbance. The
other blank was prepared in the absence of bromate-
bromide and SMT to determine the contribution of
other reactants to the absorbance of the system. Since
the absorbance of the second blank was negligible, all
absorbance measurements were made against a water
blank.
Method Validation. Analytical parameters. A lin-
ear relation is found between absorbance and concen-
tration in 1–10 µg/mL range, and Beer’s law is obeyed
in the reverse manner; the equation of the line being
Y = 0.85 + (–0.07)X,
where Y is the absorbance and X is concentration in
µg/mL. The correlation coefficient (r) of the calibration
plot is calculated to be 0.9974 confirming a linear
decrease in absorbance with concentration. The molar
absorptivity is calculated to be 3.02 × 104 L/mol cm, and
Sandel sensitivity being 0.0081 µg/cm2. The limit of
detection (LOD) and limit of quantification (LOQ) are
calculated to be 0.10 and 0.31 µg/mL, respectively.
Selectivity. To determine the selectivity of the meth-
ods, the analytical placebo was prepared and subjected
to analysis by the proposed methods. It was confirmed
that the change in absorbance with respect to the
reagent blank was caused only by the analyte. To iden-
tify the interference by common tablet excepients, a
synthetic mixture with the composition: SMT (50 mg),
talc (20 mg), starch (40 mg), gum acacia (10 mg), cal-
cium gluconate (20 mg), lactose (20 mg), sodium algi-
nate (10 mg) and magnesium stearate (10 mg), was pre-
pared and subjected to analysis by the proposed meth-
ods after solution preparation using the procedure
described for tablets. The percent recoveries of SMT
were 100.9 ± 0.7 (n = 5) and 102 ± 1 (n = 5) by titrimetry
and spectrophotometry, respectively, suggesting non-
interference by the excipients in the assay under the
described optimum conditions.
Accuracy and precision. Intra-day and inter-day
precision of the methods were assessed from the results
of seven replicate analyses of pure drug solution. The
mean and relative standard deviation (RSD) values of
replicate analyses for three different amount/concentra-
tion levels were calculated. To calculate the interday
precision, analysis was performed over a period of five
days preparing all solutions afresh each day. The accu-
racy of the methods was determined by calculating the
percentage deviation observed in the analysis of pure
drug solution and expressed as relative error (RE).
Table 1 summarises the intra-day precision and accu-
racy data for assaying SMT by the proposed methods
which are within 3%. The inter-day precision was less
than 4.5%.
2009
1196 KALSANG THARPA, KANAKAPURA BASAVAIAH

Table 1. Evaluation of intra-day and inter-day accuracy and precision

SMT Intra-day accuracy and precision Inter-day accuracy and precision

Method*
taken SMT** found RE, % RSD, % SMT** found RE, % RSD, %
Titrimetry 2.50 2.49 0.40 1.36 2.52 0.80 1.26
5.00 4.98 0.40 1.23 5.05 1.00 1.58
7.50 7.53 0.40 0.71 7.60 1.33 1.07
Spectrophotometry 2.50 2.44 2.44 1.95 2.57 2.80 2.64
5.00 4.90 1.85 1.36 5.12 2.40 2.12
7.50 7.61 1.57 1.24 7.64 1.87 1.83
Notes: * In titrimetric method, SMT taken/found, range and SD are in mg and in spectrophotometric method the same are in µg/mL.
** Mean value of seven determinations.
RE, Relative error.

Table 2. Method robustness and ruggedness (all values in % RSD)

Different acid concen- Different analysts, Different instruments

Method Nominal concentration*
trations** (n = 3 or 4) (n = 2) (n = 3)

Titrimetry 2.00 0.85 0.48 2.45

5.00 1.26 0.62 3.14
8.00 0.74 0.38 2.92
Spectrophotometry 2.00 0.62 0.22 2.64
5.00 0.44 0.36 2.37
8.00 0.72 0.54 3.52
Notes: * In titrimetry method, SMT amounts taken are in mg and in spectrophotometry method the same are in µg/mL.
** HCl acid concentrations in titrimetry method are 0.5, 0.6 and 0.7 M HCl acid concentrations in spectrophotometry method are
0.5, 1.0, 1.5 and 2.0 M HCl.

Robustness and ruggedness. For the evaluation of
the method robustness, acid concentration was slightly
varied deliberately and analysis was performed under
the altered experimental conditions. The capacity of the
methods was found to remain unaffected by small
deliberate variations. Method ruggedness was
expressed as the RSD of the same procedure applied by
two different analysts and using three different instru-
ments. The results showed no statistical difference
between different analysts and instruments suggesting
that the developed methods were rugged. The results of
the study are presented in Table 2.
Application to Tablet Assay. Twenty nine brands of
tablets containing SMT are currently available at the
Indian market. Two representative brands were assayed
and the results are presented in Table 3. The same batch
tablets were assayed by well-established literature
method [15] for comparison. The method consisted of
the measurement of absorbance of an ethanolic solution
of the drug at 238 nm. A close agreement between the
results obtained by the proposed methods and the refer-
ence method in respect of accuracy and precision was
obtained as is seen from the calculated Student’s
t-value and F-value. In a few instances the calculated
t-value and F-value exceeds the tabulated values which
can be attributed to operation of indeterminate errors.
The results also agreed with the label claim.
Recovery study. To ascertain the accuracy and valid-
ity of the proposed methods, recovery experiment was
performed via standard addition technique. To a fixed
and known amount/concentration of SMT in tablet
powder (pre-analysed), pure drug was added at three
levels (50, 100 and 150% of the level present in the tab-
let) and the total amount was found by the proposed
methods. Each experiment was repeated three times.
The recovery of pure drug added to tablet powder
ranged from 95.6 to 111% (Table 4) with a standard
deviation of 0.8–2.7 indicating that commonly added
excipients such as talc, starch, lactose, sodium alginate,
magnesium stearate, calcium gluconate, and calcium
dihydrogenorthophosphate did not interfere in the
assay.

JOURNAL OF ANALYTICAL CHEMISTRY Vol. 64 No. 11 2009

 BROMATOMETRIC ASSAY OF SIMVASTATIN 1197

Table 3. Results of assay of tablets by proposed methods

Found** (Percent of label claim ± SD)

Tablet’s* brand name Nominal amount, mg
Reference method Titrimetry Spectrophotometry

Simvofix1 10 98.6 ± 0.8 99.8 ± 1.7 97 ± 2

t = 1.51 t = 1.80
F = 4.52 F = 6.25
20 100.3 ± 0.5 99.5 ± 0.7 100.8 ± 0.6
t = 2.10 t = 1.43
F = 1.96 F = 1.44
40 102.3 ± 0.6 104 ± 2 104 ± 1
t = 2.06 t = 3.35
F = 11.11 F = 2.78
Zosta2 05 99.9 ± 0.6 99 ± 2 101 ± 1
t = 1.09 t = 2.17
F = 11.11 F = 2.78
10 101 ± 1 103 ± 0.9 100 ± 2
t = 3.32 t = 1.05
F = 1.23 F = 4.00
20 96.6 ± 0.9 95 ± 2 97.9 ± 2
t = 1.74 t = 1.41
F = 4.94 F = 4.94
1 2
Notes: * Marketed by: Bal Pharma (Servetus), USV (Corvette).
** Mean value of five determinations.
Tabulated t-value at the 95% confidence level is 2.77.
Tabulated F-value at the 95% confidence level is 6.39.

CONCLUSION reported procedures [16] which require a heating step.

This is a first report on the assay of simvastatin by
titrimetry and the third one by visible spectrophotome-
try. The demonstrated methods for the quantification of
simvastatin in bulk drug and in tablets are simple, rapid
and accurate. The methods are of wider applicability
than those reported earlier. Compared to the existing
spectrophotometric methods, the proposed method has
the advantages of wide linear dynamic range, high sen-
sitivity and milder experimental conditions without
heating or extraction step. The relative cheapness of the
techniques, besides sensitivity, demonstrates their
advantageous features. In fact, the spectrophotometric
method excels the reported UV-spectrophotometric
methods which have narrow linear ranges, 4–12 µg/mL
[13], 12–28 µg/mL [14] and 2–16 µg/mL [15], in sen-
sitivity. The method has been found to be superior to a
couple of HPLC methods which are applicable over
2−200 µg/mL [3] and 0.5–5.0 µg/mL [6] concentration
ranges. The present spectrophotometric method
employes simple working conditions unlike the
The acid concentration used is low (0.5 M HCl overall)
whereas the method of Vijaya saradhi et al. [17]
requires concentrated HNO3 as the reaction medium.
The latter method is less sensitive with working con-
centration range of 10–50 µg/mL and an ∈ value of
3.2 × 103 L/mol cm whereas the present method can be
employed over a wide concentration range
(1−10 µg/mL) and at the same time is ten times more
sensitive (∈ = 3.02 × 104 L/mol cm) than the reported
method [17]. The titrimetric method is applicable over
the micro range (1–10 mg) but reaction time was some-
what critical. Both the methods can be used for the
rapid assay of simvastatin-containing pharmaceuticals.
ACKNOWLEDGEMENT
Authors thank M/S. Jubiliant Organosis, Mysore,
for gifting pure simvastatin. One of the authors (Kal-
sang Tharpa) thanks the authorities of the University of
Mysore, Mysore, for permission and facilities. Kalsang
Tharpa also thanks the Department of Education, Cen-

JOURNAL OF ANALYTICAL CHEMISTRY Vol. 64 No. 11 2009

1198 KALSANG THARPA, KANAKAPURA BASAVAIAH

Table 4. Results of recovery experiment

Tablet studied Method SMT in tablet Pure SMT added Total found Pure SMT recovered (Percent ± SD)

Simvofix 40 mg Titrimetry 2.60 1.50 4.15 103 ± 2

2.60 3.00 5.66 102 ± 1
2.60 4.50 7.15 101 ± 1
Spectrophotometry 2.59 1.5 4.26 111 ± 2
2.59 3.0 5.50 97 ± 2
2.59 4.5 7.06 99 ± 1
Zosta 10 mg Titrimetry 2.57 1.5 4.10 102 ± 2
2.57 2.3 5.44 96 ± 1
2.57 4.5 7.18 102 ± 1
Spectrophotometry 2.50 1.5 4.05 103 ± 3
2.50 3.0 5.52 101 ± 2
2.50 4.5 7.05 101 ± 1
In titrimetric method, SMT in tablet, pure SMT added, total SMT found are in mg.
In spectrophotometric method, SMT in tablet, pure SMT added, total SMT found are in µg/mL.

tral Tibetan Administration of His Holiness the Dalai
Lama, for providing research scholarship.
REFERENCES
1. McEvoy, G.K., HFS Drug Information, American Soci-
ety of Health System Pharmasists, 2002.
2. The United States Pharmacopoeia 25, United States
Pharmocopocial Convention, 2002, p. 9571.
3. Ying, L. and Jianwei, X., Yaowu Fenxi Zazhi, 2005,
vol. 25, p. 523.
4. AbuNameh, E.S.M., Shawabkeh, R.A., and Ali, A.,
Zhurn. anal. khim., 2006, vol. 61, no. 1, p. 70 [J. Anal.
Chem. (Engl. Transl.], vol. 61, no. 1, p. 63].
5. Ricardo, G., Gloria, G.C., Marta, D.D., and Carolina, G.,
J. Chilean Chem. Soc., 2004, vol. 49, p. 289.
6. Yan, X., Cao, G., He, X., Hu, X., and Gu, D., Huaxi
Yaoxue Zazhi, 2000, vol. 15, p. 205.
12. Wang, L. and Asgharnejad, M., J. Pharm. Biomed. Anal.,
2000, vol. 21, p. 1243.
13. Li, Z. and Jang, S., Thongguo Yaoxue Zazhi, 2000,
vol. 35, p. 554, Sci. Finder, CAN 133: 301278, AN 2000:
675973.
14. Erk, N., Pharmazie, 2002, vol. 57, p. 817.
15. Xu, L., Thongguo Yiyao Gongye Zazhi, 2001, vol. 32,
p. 271, Sci. Finder, CAN 135: 362691, AN 2001: 606960.
16. Saradhi, S.V., Himabindu, V., and Rao, G.V., Acta Cien-
cia Indica, 2007, vol. 33, p. 205.
17. Vijaya, S.S., Devala Rao, G., Sitaram, B., Pravallika, K.,
Amulya, C.P., and Sindhu, V., Int. J. Chem. Sci., 2008,
vol. 6, p. 269.
18. Basavaiah, K. and Anil Kumar, U.R., J. Mexican Chem.
Soc., 2007, vol. 51, p. 106.
19. Basavaiah, K. and Anil Kumar, U.R., Bulgarian Chem.
Commun., 2007, vol. 39, p. 159.

7. Wang, J., Thongguo Yiyao Gongye Zazhi, 2000, vol. 31,
p. 121, Sci. Finder, CAN 132: 284323, AN 2000: 293986.
8. Carlcucci, G. and Mazzeo, P., Il Farmaco, 1992, vol. 47,
p. 817.
9. Chaudhari, B.G., Patel, N.M., and Shah, P.B., J. AOAC
Int., 2007, vol. 90, p. 1242.
10. Coruh, O. and Ozkan, S.A., Die Pharmazie, 2006,
vol. 61, p. 285.
11. Srinivasu, M.K., Raju, A.N., and Reddy, G.O., J. Pharm.
Biomed. Anal., 2002, vol. 29, p. 715.
20. Basavaiah, K., Ramakrishna, V., and Anil Kumar, U.R.,
Proc. Ind. Nat. Sci. Acad., 2007, vol. 73, p. 121.
21. Basavaiah, K. and Anil Kumar, U.R., Ind. J. Chem. Tech-
nol., 2007, vol. 14, p. 611.
22. Basavaiah, K. and Anil Kumar, U.R., Ecletica Quimica,
2007, vol. 4, p. 154.
23. Basavaiah, K. and Anil Kumar, U.R., e-J. Chem., 2006,
vol. 3, p. 242.
24. Sandel, E.B., Colorimetric Determination of Trace of
Metals, New York: Interscience, 1959.

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