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Asian J. Research Chem. 2(1): Jan.

-March, 2009
,

ISSN 0974-4169 www.ajrconline.org

RESEARCH ARTICLE

Simultaneous UV Spectrophotometric Method for Estimation of Losartan


Potssium and Amlodipine Besylate in Tablet Dosage Form.
Priyanka R Patil*, Sachin U Rakesh, PN Dhabale, and KB Burade
Govt. College of Pharmacy, Karad- 415 124, (Satara), Maharashtra, India
*Corresponding Author E-mail: priyapatil5586@rediffmail.com

ABSTRACT
Two simple, accurate, precise, reproducible, requiring no prior separation and economical procedures for simultaneous
estimation of Losartan Potassium and Amlodipine Besylate in tablet dosage form have been developed. First method
employs formation and solving of simultaneous equation using 208 nm and 237.5 nm as two analytical wavelengths
for both drugs in methanol. The second method is Q value analysis based on measurement of absorptivity at 242.5 nm
(as an iso-absorptive point) and 237.5 nm. Losartan Potassium and Amlodipine Besylate at their respective max 208
nm and 237.5 nm and at isoabsorptive point 242.5 nm shows linearity in a concentration range of 2-20 g/mL.
Recovery studies range from 99.95% for Losartan Potassium and 99.33% for Amlodipine Besylate in case of
simultaneous equation method and 102.93% for Losartan Potassium and 101.02% for Amlodipine Besylate in case of
Q - analysis method confirming the accuracy of the proposed method. The proposed method is recommended for
routine analysis since it is rapid, simple, accurate and also sensitive and specific by no heating and no organic solvent
extraction.

KEY WORDS: Losartn Potassium, Amlodipine Besylate, max , Simultaneous equation method, Q analysis

INTRODUCTION: They include high performance liquid chromatography,12-17


Losartan (I, 2-n-butyl-4-chloro-1-[p-(o-1H-tetrazol- 5- reversed phase high performance liquid chromatography,18-
21
ylphenyl) benzyl]-imidazole-5methanol monopotassium salt) high performance thin layer chromatography,22-25 gas
is a highly selective, orally active, non-peptide angiotensin II chromatography,26 gas chromatography–mass
receptor antagonist indicated for the treatment of spectrometry,27
hypertension. It has a more potent active metabolite liquid chromatography with tandem mass spectrometry28
EXP3174 (II, 2-n-butyl-4-chloro-1-[2-(1H-tetrazol-5 yl) and fluorimetry,29 derivative spectroscopy,30,31
biphenyl- 4-yl) methyl] imidazole-5-carboxyl acid)1. The simultaneous multicomponent mode of analysis and
determination of Losartan has been carried out in tablets by difference spectrophotometry32-34.
HPLC, capillary electrophoresis and super-critical fluid
chromatography2,3, in urine by gas chromatography- mass By these two methods no UV spectrophotometric study
spectrometry4 and, simultaneously with its active metabolite on Losartan and Amlodipine in tablet dosage form in
in biological fluids, by HPLC5- 10. pharmaceutical preparations has been found in literature
survey. There was only one method has been reported35
Amlodipine, chemically, 2-[(2- aminoethoxy) methyl]- 4- for estimation of Losartan and Amlodipin in tablet by
(2-chlorophenyl) -1, 4-dihydro- 6-methyl-3, 5- absorption correction method, which prompted to pursue
pyridinedicarboxylic acid 3-ethyl, 5-methyl ester, is an anti- the present work. The objective of the present work is to
hypertensive and an antianginal agent in the form of the develop and validate new analytical methods for
besylate salt, Amlodipine besylate. It is not official in any simultaneous determination of Losartan Potassium and
Pharmacopoeia. Various analytical methods have been Amlodipine Besylate in tablet dosage form. This
reported for the assay of Amlodipine besylate11 in pure communication forms the first report of two simple,
form as well as in pharmaceutical formulations. sensitive and reproducible methods for the simultaneous
estimation of Losartan Potassium and Amlodipine
Besylate from combined dosage form.

MATERIALS AND METHODS:


Received on 20.02.2009 Modified on 13.04.2009 Materials:
Accepted on 24.05.2009 © AJRC All right reserved Spectral runs were made on a Shimadzu UV-Visible
Asian J. Research Chem. 2(2): April.-June, 2009 page 183-187 spectrophotometer, model- 1700 (Japan) was employed
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Asian J. Research Chem. 2(1): Jan.-March, 2009
,
with spectral bandwidth of 0.5 nm and wavelength A1 a x2 – A2 a x1
accuracy of ± 0.3 nm with automatic wavelength Cy = a y1ax2 - ay2a x1 …………Eq. (ii)
corrections with a pair of 10 mm quartz cells. Glasswares
used in each procedure were soaked overnight in a Where, A1 and A2 are absorbances of mixture at 208 nm
mixture of chromic acid and sulphuric acid rinsed and 237.5 nm respectively, ax1 and ax2 are absorptivities
thoroughly with double distilled water and dried in hot air of LP at 1 and 2 respectively and ay1 and ay2 are
oven. Amlodipine besylate reference standard was kindly absorptivities of AB at 1 and 2 respectively. Cx and Cy
provided by Shreya Life Sciences Pvt. Ltd. Aurangabad are concentrations of LP and AB respectively.
(M. S.) while Losartan Potassium was provided by Lupin
Research Park, Pune. The pharmaceutical preparations of Method II (Absorbance ratio or Q-analysis method):
combination of Losartan and Amlodipine that is Alsartan From the overlain spectrum of LP and AB, two
AM tablet (ARISTO, Mumbai). Methanol of analytical wavelengths were selected one at 242.5 nm which is the
reagent grade was purchased by Loba Chemie Pvt. Ltd. isoabsorptive point for both the drugs and the other at
(India). All the solutions were protected for light and were 237.5 nm which is max of AB . The absorbances of the
analyzed on the day of preparations. sample solutions prepared in a similar manner as in the
previous method, were measured and the absorptivity
Selection of common solvent: values for both drugs at the selected wavelengths were
Methanol of analytical reagent grade was selected as also calculated. The method employs Q values and the
common solvent for developing spectral characteristics concentrations of drugs in sample solution were
of drug. The selection was made after assessing the determined by using the following formula,
solubility of both the drugs in different solvents. For LP
Q0 – Q2 a1
Preparation of Standard Drug Solution: C1 = ×
Standard stock solutions containing Losartan Potassium Q1–Q2 A
(LP) and Amlodipine besylate (AB) were prepared For AB
individually by dissolving 2.5 mg of LP and quantity of Q0 – Q1 a2
AB equivalent to Amlodipine base 2.5 mg separately in C2 = ×
20 ml of methanol. It was then sonicated for 10 minutes Q2–Q1 A
and the final volume of both the solutions were made up Where,
to 50 ml with methanol to get stock solutions containing Absorbance of sample at 237.5 nm
50 g/ mL each of LP and AB in two different 50 ml Q0 =
volumetric flasks. Absorbance of sample at 242.5 nm

Determination of Absorption Maxima: Absorptivity of LP at 237.5 nm


By appropriate dilution of two standard drug solutions Q1 =
with methanol, solutions containing 10 g ml-1 of LP Absorptivity of LP at 242.5nm
and 10 g ml-1 of AB were scanned separately in the
range of 200- 400 nm to determine the wavelength of Absorptivity of AB at 237.5 nm
maximum absorption for both the drugs. LP and AB Q2 =
showed absorbance maxima at 208 nm ( 1) and 237.5 Absorptivity of AB at 242.5nm
nm ( 2) respectively. The overlain spectra showed max
of both drugs and also isoabsorptive points at 242.5 nm A = Absorbance of sample at isoabsorptive point,
(Fig. 1). a1 and a2 = Absorptivities of LP and AB respectively at
isoabsorptive point.
Method I (Simultaneous equation method):
Two wavelengths selected for the method are 208 nm
and 237.5 nm that are absorption maximas of LP and AB Application of the proposed method for the
respectively in methanol. The stock solutions of both determination of LP and AB in tablets:
the drugs were further diluted separately with methanol Twenty tablets of marketed formulation Alsartan AM
to get a series of standard solutions of 2-20 g /mL (ARISTO, Mumbai) containing LP 50 mg and AB
concentrations. The absorbances were measured at the equivalent to Amlodipine base 5 mg were weighted, and
selected wavelengths and absorptivities (A 1%, 1 cm) finely powdered. For analysis of drug, a standard addition
for both the drugs at both wavelengths were determined method was used. An accurately weighted 2.5 mg of pure
as mean of three independent determinations. AB was added to finely powdered samples to bring the
Concentrations in the sample were obtained by using concentration of AB in linearity range. With this addition, the
following equations- ratio of LP and AB in the samples was brought to 1:1.
A1 ay2 – A2 a y1 Quantity of powder equivalent to 5 mg of LP and 5 mg of
Cx = …………Eq. (i) Amlodipine base was weighed and dissolved in 40 mL of
ax1ay2-ax2ay1 methanol and sonicated for 10 minutes. Then the solution

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Table 1: Linear regression analysis of calibration curves with their respective absorptivity values.
Parameter Method I Method II
LP AB LP AB
Beer’s law limit ( g ml-1) 2-20 2-20 2-20 2-20
Correlation coefficient (r) 0.9988 0.9991 0.9993 0.9970
Molar absorptivity (lit/mole/cm) 42942.37 19435.23 21396.44 18694.30
Sandell's sensitivity 0.0107 0.0210 0.021546 0.021872
(mcg/Sq.cm/0.001)
Slope 0.0955 0.0468 0.04602 0.04325
Intercept -0.0063 -0.0158 -0.017533 -0.02353

Table 2: Results of analysis of laboratory samples.


Analyte Method I Method II
LP AB LP AB
% Conc. Estimated (Mean ± S.D.) 99.79 ± 0.13 99.74 ± 0.15 99.85 ± 0.03 101.09 ± 0.82
Coefficient of variance 0.0008 0.0002 0.0001 0.0004
Average of three determinations; R.S.D.; Relative Standard Deviation.

Table 3: Results of analysis of tablet samples.

Method Drug Label Claim % Label Claim ± R. S. D. Coefficient of variance % Recovery* (Mean ±
R. S. D)
LP 5 103.39 ± 1.224 0.0155 99.95± 0.97
I AB 50 105.06± 3.25 0.111 99.33± 0.72
LP 5 103.93± 2.66 0.072 102.93± 0.21
II AB 50 105.05± 3.25 0.111 101.02± 0.37
Average of three determinations; R.S.D.; Relative Standard Deviation
Table 4: Results of intermediate precisions.
Method I Method II
Day % Label claim estimated % Label claim estimated
(Mean ± % R.S.D.) (Mean ± % R.S.D.)
LP AB LP AB
Intraday 105.83 100.04 100.87 112.5
± 0.73 ± 0.43 ± 0.56 ± 2.2
Interday 106.86 108.05 99.25 100.6
± 1.27 ± 1.71 ± 1.76 ± 0.36
.

was filtered through whatman filter paper no. 41 and then Accuracy:
final volume of the solution was made up to 50 ml with To ascertain the accuracy of the proposed methods,
methanol to get a stock solution containing 100 g ml-1of LP recovery studies were carried out by standard addition
and 100 g ml-1 AB. Appropriate aliquots of LP and AB method at three different levels (80%, 100% and 120%).
within the Beer’s law limit were taken. In Method I, the Percent recovery for LP and AB, by both the methods,
concentration of both LP and AB were determined by was found in the range of 101.44% to 100.17%.
measuring the absorbance of the sample at 208 nm and 237.5
nm. Values were substituted in the respective formula to Linearity:
obtain concentrations. The linearity of measurement was evaluated by
analyzing different concentration of the standard
For Method II, the concentration of both LP and AB solution of LP and AB. For simultaneous equation
were determined by measuring absorbance of the sample method and Q analysis, the Beer- Lambert’s
at 242.5 nm and 237.5 nm and values were substituted in concentration range was found to be 2-20 g/ml for LP
the respective formula to obtain concentrations. Results and AB.
of tablet analysis are shown in Table 3.
Precision:
VALIDATION: Precision was studied to find out intra and inter-day
The method was validated according to ICH Q2B variations in the test method of LP and AB. Calibration
guidelines for validation of analytical procedures in order to curves prepared in medium were run in triplicate in same day
determine the linearity, sensitivity, precision and accuracy and for three days. %RSD (relative standard deviation) were
for the analyte.
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calculated which should be less than 2 %. The results are respectively. We are also thankful to Mr. Shreeniwas
tabulated in Table 4. Mohite sir for his moral support and guidance.

Fig.1: Overlain spectra of LP and AB. Overlain spectra of REFERENCES:


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