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:[email protected] 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
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-
ylphenyl) benzyl]-imidazole-5methanol monopotassium salt)
21 high performance thin layer chromatography,22-25 gas
is a highly selective, oral y active, non-peptide angiotensin II
receptor antagonist indicated for the treatment of
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)
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
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
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
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
MATERIALS AND METHODS:
Received on 20.02.2009 Modified on 13.04.2009
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
Asian J. Research Chem. 2(1): Jan.-March, 2009
with spectral bandwidth of 0.5 nm and wavelength
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
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.
Preparation of Standard Drug Solution:
Standard stock solutions containing Losartan Potassium
(LP) and Amlodipine besylate (AB) were prepared
individually by dissolving 2.5 mg of LP and quantity of
AB equivalent to Amlodipine base 2.5 mg separately in
20 ml of methanol. It was then sonicated for 10 minutes
and the final volume of both the solutions were made up
to 50 ml with methanol to get stock solutions containing
50 g/ mL each of LP and AB in two different 50 ml
Determination of Absorption Maxima:
By appropriate dilution of two standard drug solutions
with methanol, solutions containing 10 g ml-1 of LP
and 10 g ml-1 of AB were scanned separately in the
range of 200- 400 nm to determine the wavelength of
maximum absorption for both the drugs. LP and AB
showed absorbance maxima at 208 nm ( 1) and 237.5
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,
a1 and a2 = Absorptivities of LP and AB respectively at
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
ratio of LP and AB in the samples was brought to 1:1.
Quantity of powder equivalent to 5 mg of LP and 5 mg of
Amlodipine base was weighed and dissolved in 40 mL of
methanol and sonicated for 10 minutes. Then the solution
Asian J. Research Chem. 2(1): Jan.-March, 2009 Table 1: Linear regression analysis of calibration curves with their respective absorptivity values. Parameter Method II Table 2: Results of analysis of laboratory samples. Method II % Conc. Estimated (Mean ± S.D.) Coefficient of variance
Average of three determinations; R.S.D.; Relative Standard Deviation.
Table 3: Results of analysis of tablet samples. Label Claim % Label Claim ± R. S. D. Coefficient of variance % Recovery* (Mean ±
Average of three determinations; R.S.D.; Relative Standard Deviation
Table 4: Results of intermediate precisions. Method II % Label claim estimated % Label claim estimated (Mean ± % R.S.D.) (Mean ± % R.S.D.) Intraday Interday
. 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
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
of tablet analysis are shown in Table 3.
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
Asian J. Research Chem. 2(1): Jan.-March, 2009
calculated which should be less than 2 %. The results are
respectively. We are also thankful to Mr. Shreeniwas
Mohite sir for his moral support and guidance.
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