Bromato Simvastatin PDF
Bromato Simvastatin PDF
Bromato Simvastatin PDF
, 2009.
ARTICLES
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
1193
1194 KALSANG THARPA, KANAKAPURA BASAVAIAH
measurable at 730 nm. In the other two procedures, the was weighed into a 100 mL calibrated flask, 60 mL of
iron (II) formed is chelated with 1,10-phenanthroline or 3 : 2 acetic acid were added and the content was shaken
2,2'-bipyridine followed by measurement of absor- for 15–20 min, diluted to the mark with same acid,
bance at 480 or 490 nm. Very recently a procedure mixed well and filtered using a Whatman no. 42 filter
based on the formation of yellow colour on binding paper. First 10 mL portion of the filtrate was discarded
SMT with ferric nitrate in concentrated nitric acid has and a convenient aliquot (say 5 mL) of the subsequent
been reported by Vijaya et al. [17]. However, literature portion of the filtrate was assayed by titrimetry. The
survey revealed that no titrimetric methods have ever same tablet extract was appropriately diluted with 3 : 2
been reported for the determination of SMT. During the acetic acid before analysis by spectrophotometry.
preliminary studies, SMT was found to react with Procedures. Titrimetry. A 10 mL aliquot of standard
in situ generated bromine in HCl medium which drug solution containing 1–10 mg of SMT was accu-
prompted the authors to develop titrimetric and spectro- rately measured and transferred into a 100 mL Erlenm-
photometric methods for the determination of drug eyer flask. The solution was acidified by adding 5 mL
based on this reaction. This paper describes the study of 2 M HCl and 1 mL of glacial acetic acid. 10 mL of
and evaluation of the variables which govern the inter- bromate-bromide mixture (2 mM in KBrO3) was added
action of SMT and in situ bromine in order to provide to the flask by means of a pipette with swirling and the
the basis for the simple, sensitive and cost-effective tit- flask was stoppered and kept aside for 2–3 min with
rimetric and spectrophotometric methods for the deter- occasional swirling. Then, 5 mL of 10% KI solution
mination of SMT. The methods use bromate-bromide was added and the liberated iodine titrated with
mixture in HCl medium as the brominating agent and 0.012 M thiosulphate to a starch end point. A blank
thiocyanate as the complexing agent. The proposed titration was performed taking 10 mL of 3:2 acetic acid
methods have the advantage of accuracy and precision in place of drug solution. The amount of SMT in the ali-
besides being free from interference from common tab- quot was calculated from:
let excipients.
VSMw
SMT ( mg ) = ------------------ ,
MATERIALS AND METHODS 0.666
Apparatus. A systronics model 106 digital spectro- where V - KBrO3 reacted, mL, S - Strength of KBrO3 in
photometer provided with 1-cm matched quartz cells M and Mw - relative molecular mass of SMT.
was used for absorbance measurements. Spectrophotometry. Different aliquots (0, 0.25, 0.5,
Reagents. All chemicals used were of analytical 1.0, 1.5, 2.0, 2.5 mL) of standard 40 µg/mL SMT were
reagent grade and distilled water was used throughout accurately transferred into a series of 10 mL calibrated
the study. A standard stock solution of bromate-bro- flasks. To each flask, 1mL of 5 M HCl was added fol-
mide equivalent to 2 mM KBrO3–20 mM KBr was pre- lowed by 1 mL of bromate-bromide (30 µg/mL in
pared by dissolving accurately weighed 0.3340 g of KBrO3). The flasks were stoppered, content was mixed
KBrO3 and 2.38 g of KBr in water and diluting to vol- and kept aside for 10 min with occasional swirling.
ume in a one-litre volumetric flask, and used in titrime- Then, 1mL of FAS (600 µg/mL) was added to each
try. A stock standard solution of KBrO3−KBr equivalent flask, and after 5 min, 1 mL of 3 M thiocyanate was
to 300 µg/mL KBrO3 was prepared by dissolving accu- added and diluted to the mark with water. The absor-
rately weighed 30 mg of KBrO3 and 300 mg of KBr in bance of each solution was measured at 470 nm against
water and diluting to mark in a 100 mL calibrated flask. a water blank. A standard graph was prepared by plot-
It was diluted with water to obtain a working concen- ting the measured absorbance Vs concentration (µg/mL)
tration of 30 µg/mL in KBrO3. Hydrochloric acid (2 and of SMT. The concentration of the unknown was read
5 M), sodium thiosulphate (0.012 M), potassium thio- from the calibration graph or computed from the
cyanate (3 M), KI (10%) and starch indicator (1%) regression equation derived using the Beerís law data.
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 RESULTS AND DISCUSSION
of dilute H2SO4 and diluted to volume with water in Potassium bromate in the presence of a large excess
250 mL calibrated flask. A stock standard solution of of bromide in acid medium is valuable oxidimetric/bro-
drug containing 1 mg/mL SMT was prepared by dis- minating agent and has been successfully employed in
solving accurately weighed pure SMT in 3 : 2 acetic the assay of many drug substances [18–23]. The present
acid and diluting with the same acid, and used in titri- communication deals with the titrimetric and spectro-
metry. For spectrophotometric work, the stock solution photometric assay of SMT using in situ generated bro-
was diluted to 40 µg/mL with 3 : 2 acetic acid. mine as a brominating agent. The proposed methods are
Tablets containing SMT were purchased from local indirect and are based on the determination of residual
commercial sources for investigation. Twenty tablets bromine after the reaction between the drug and bro-
were weighed accurately and finely powdered. An mine is judged to be and rely on different reaction
amount of the powder equivalent to 100 mg of SMT schemes.
Method development. Titrimetry. Direct titration used in the study. The bromination step took 5 min
of SMT with bromate-bromide was not successful. while the reduction and subsequent complexation steps
Assay by back titration was found feasible, and hence were instantaneous. Developed colour was stable for at
several experimental variables were optimized to get least 60 min in the presence of the reaction product.
accurate and precise results. Accurate and reproducible Two blanks were prepared in this study. The reagent
stoichiometry was obtained when 2 M HCl concentra- blank which contained optimum concentrations of all
tion was maintained in addition to acetic acid present in reactants except SMT gave maximum absorbance. The
the drug solution. Hence, 5 mL of 2 M HCl in a total other blank was prepared in the absence of bromate-
volume of 15 mL (0.66 M overall) were used in the bromide and SMT to determine the contribution of
study. 1 mL of glacial acetic acid was found necessary other reactants to the absorbance of the system. Since
to clear little turbidity that was formed on adding HCl. the absorbance of the second blank was negligible, all
The bromination reaction was found to be complete in absorbance measurements were made against a water
2–3 min and it is essential to terminate the reaction at blank.
the end of the 3rd min but not before the 2nd min after Method Validation. Analytical parameters. A lin-
adding bromate-bromide mixture. Beyond 3 min and ear relation is found between absorbance and concen-
upto 60 min little more bromine was found to be con- tration in 1–10 µg/mL range, and Beer’s law is obeyed
sumed but without yielding a definite reaction stoichi- in the reverse manner; the equation of the line being
ometry. Hence, the reaction time was stricted to
2−3 min. For the range investigated (1–10 mg) the reac- Y = 0.85 + (–0.07)X,
tion yielded a 3 : 2 stoichiometry (SMT : KBrO3). Out- where Y is the absorbance and X is concentration in
side this range and reaction time, non-stoichiometric µg/mL. The correlation coefficient (r) of the calibration
results were obtained. plot is calculated to be 0.9974 confirming a linear
Spectrophotometry. Complex formation reaction decrease in absorbance with concentration. The molar
involving iron (III) and thiocyanate has been the basis absorptivity is calculated to be 3.02 × 104 L/mol cm, and
for the sensitive determination of the former in a variety Sandel sensitivity being 0.0081 µg/cm2. The limit of
of matrices [24]. This reaction in combination of bro- detection (LOD) and limit of quantification (LOQ) are
mate-bromide mixture has been applied for the assay of calculated to be 0.10 and 0.31 µg/mL, respectively.
some pharmaceuticals [18–20]. The present method is Selectivity. To determine the selectivity of the meth-
based on the bromination of SMT by a measured excess ods, the analytical placebo was prepared and subjected
of in situ generated bromine in acid medium, reduction to analysis by the proposed methods. It was confirmed
of residual bromine by fixed amount of iron (II) and that the change in absorbance with respect to the
subsequent complexation of iron (III) formed with thio- reagent blank was caused only by the analyte. To iden-
cyanate which is measured at 470 nm. When a fixed tify the interference by common tablet excepients, a
concentration of in situ generated bromine reacts with synthetic mixture with the composition: SMT (50 mg),
increasing concentration of SMT, there occurs a con- talc (20 mg), starch (40 mg), gum acacia (10 mg), cal-
comitant fall in the bromine concentration. When the cium gluconate (20 mg), lactose (20 mg), sodium algi-
unreacted bromine is reduced by a fixed concentration nate (10 mg) and magnesium stearate (10 mg), was pre-
of iron (II), there will be a proportional decrease in the pared and subjected to analysis by the proposed meth-
concentration of iron (III). This is observed as a propor- ods after solution preparation using the procedure
tional decrease in the absorbance of iron (III)-thiocyn- described for tablets. The percent recoveries of SMT
ate complex with increase in SMT concentration, were 100.9 ± 0.7 (n = 5) and 102 ± 1 (n = 5) by titrimetry
which formed the basis for the assay of drug. and spectrophotometry, respectively, suggesting non-
interference by the excipients in the assay under the
The conditions for the determination of iron (III) described optimum conditions.
with thiocyanate are well established [24]. Hence, var-
Accuracy and precision. Intra-day and inter-day
ious parameters associated with the bromination of
precision of the methods were assessed from the results
drug, and subsequent reduction of residual bromine by
of seven replicate analyses of pure drug solution. The
iron (II) where optimized. 1 mL of 5 M HCl in a total
mean and relative standard deviation (RSD) values of
volume of about 5 mL for bromination and reduction
replicate analyses for three different amount/concentra-
steps was used, and the same quantity of HCl in a total
tion levels were calculated. To calculate the interday
of 10 mL was used for the complexation step.
precision, analysis was performed over a period of five
Considering 8 µg/mL as the upper limit of iron (III) days preparing all solutions afresh each day. The accu-
that could be accurately determined by thiocyanate racy of the methods was determined by calculating the
method, 30 µg/mL KBrO3 in the presence of a large percentage deviation observed in the analysis of pure
excess of bromide was found to generate it from drug solution and expressed as relative error (RE).
56 µg/mL FAS under the specified conditions. How- Table 1 summarises the intra-day precision and accu-
ever, a slight excess (60 µg/mL) FAS was used to ensure racy data for assaying SMT by the proposed methods
a quantitative reaction. Hence, 1 mL each of 30 µg/mL which are within 3%. The inter-day precision was less
KBrO3 (with excess of KBr) and 600 µg/mL FAS were than 4.5%.
Robustness and ruggedness. For the evaluation of obtained as is seen from the calculated Student’s
the method robustness, acid concentration was slightly t-value and F-value. In a few instances the calculated
varied deliberately and analysis was performed under t-value and F-value exceeds the tabulated values which
the altered experimental conditions. The capacity of the can be attributed to operation of indeterminate errors.
methods was found to remain unaffected by small The results also agreed with the label claim.
deliberate variations. Method ruggedness was
expressed as the RSD of the same procedure applied by Recovery study. To ascertain the accuracy and valid-
two different analysts and using three different instru- ity of the proposed methods, recovery experiment was
ments. The results showed no statistical difference performed via standard addition technique. To a fixed
between different analysts and instruments suggesting and known amount/concentration of SMT in tablet
that the developed methods were rugged. The results of
the study are presented in Table 2. powder (pre-analysed), pure drug was added at three
levels (50, 100 and 150% of the level present in the tab-
Application to Tablet Assay. Twenty nine brands of let) and the total amount was found by the proposed
tablets containing SMT are currently available at the methods. Each experiment was repeated three times.
Indian market. Two representative brands were assayed The recovery of pure drug added to tablet powder
and the results are presented in Table 3. The same batch ranged from 95.6 to 111% (Table 4) with a standard
tablets were assayed by well-established literature
method [15] for comparison. The method consisted of deviation of 0.8–2.7 indicating that commonly added
the measurement of absorbance of an ethanolic solution excipients such as talc, starch, lactose, sodium alginate,
of the drug at 238 nm. A close agreement between the magnesium stearate, calcium gluconate, and calcium
results obtained by the proposed methods and the refer- dihydrogenorthophosphate did not interfere in the
ence method in respect of accuracy and precision was assay.
Tablet studied Method SMT in tablet Pure SMT added Total found Pure SMT recovered (Percent ± SD)
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