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Asian J. Research Chem. 2(1): Jan.-March, 2009 ISSN 0974-4169
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]


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


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 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 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.
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.)

was filtered through whatman filter paper no. 41 and then
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:
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:
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. Fig.1: Overlain spectra of LP and AB. Overlain spectra of
Losartan potassium (RM) and Amlodipine besylate (AB) in
1. Michelle Polinko, Kerry Riffel, Hengchang Song, Man- methanol.
Wai Lo. Simultaneous determination of losartan and EXP3174 in human plasma and urine utilizing liquid chromatography/ tandem mass spectrometry. J Pharma 2. Williams R.C.; Alasandro M.S.; Fasone V.L.; Boucher R.J.; Edwards J.F. Comparison of liquid chromatography, capillary electrophoresis and super-critical fluid chromatography in the determination of Losartan Potassium drug substance in Cozaar tablets. J Pharma 3. Mccarthy K. E, Qingxi Wang, Tsai E. W. Gilbert R. E. Ip D. P. Brooks M. A. Determination of losartan and its degradates in COZAAR tablets by reversed-phase high- performance thin-layer chromatography. J Pharm Biomed RESULTS AND DISCUSSION:
The overlain spectra of LP and AB exhibit max of 208 4. H.H. Maurer, T. Kraemer, J.W. Arlt Screening for the nm and 237.5 nm for LP and AB respectively which are Detection of Angiotensin-Converting Enzyme Inhibitors, quite separated from each other. Additionally one Their Metabolites, and AT II Receptor Antagonists. Ther. isoabsorptive point was observed at 242.5 nm. This 5. C.I. Furtek, M. W. Lo. Simultaneous determination of a wavelength was selected for simultaneous estimation of novel angiotensin II receptor blocking agent, losartan, and LP and AB for Q value analysis and it is assume to be its metabolite in human plasma and urine by high- sensitive wavelength. Standard calibration curves for LP performance liquid chromatography. J Chroma: Biomed and AB were linear with correlation coefficients (r) values in the range of 0.9970- 0.9993 at all the selected 6. H. Lee, H.O. Shim, H.S. Lee. Simultaneous wavelengths and the values were average of three Determination of Losartan and Active Metabolite readings with standard deviation in the range of 0.13 – EXP3174 in Rat Plasma by HPLC with Column 0.82. The calibration curves were repeated three times in Switching H. Lee 1 / H. O. Shim 1 / H. S. Lee 2. a day and the average % RSD was found to be 0.645 for 7. A.F.M. El Walily, S.F. Belal, E.A. Heaba, et al. An LP and 1.31 for AB, similarly the method was repeated improved method for the simultaneous determination of for three different days and average % RSD was found losartan and its major metabolite, EXP3174, in human to be 1.51 for LP and 1.03 for AB. The accuracy of the plasma and urine by high-performance liquid method was conformed by recovery studies from tablet chromatography with fluorescence detection J Pharma at three different levels of standard additions; recovery in the range of 95 – 110% justifies the accuracy of 8. Farthing D, Sica D, Fakhry I, Pedro A, Gehr TWB. Simple high-performance chromatographic method for determination of losartan and E-3174 metabolite in CONCLUSION:
human plasma, urine and dialysate. J Chromatogr B 1997; The most striking feature of this method is its simplicity 9. Andrea Soldner, Hildegard Spahn-Langguth, Ernst and rapidity, non- requiring- consuming sample Mutschler. HPLC assays to simultaneously determine the preparations such as extraction of solvents, heating, angiotensin-AT1 antagonist losartan as well as its main and degassing which are needed for HPLC procedure. These active metabolite EXP 3174 in biological material of are new and novel methods and can be employed for routine analysis in quality control analysis. The described J Pharma Biomed Anal. 1998; 16 (5): 863-873. methods give accurate and precise results for 10. Andrea Soldner, Hildegard Spahn-Langguth, Dieter Palm, determination of Losartan potassium and Amlodipine Ernst Mutschler. A radioreceptor assay for the analysis of besylate mixture in marketed formulation. complementary LC data reveals a potential contribution of active metabolites. J Pharma Biomed Anal. 1998; 17 ACKNOWLEDGEMENTS:
The authors are thankful to the Principal Dr. S. B. Bhise, 11. Argekar A P, Powar S G. Simultaneous determination of Govt. College of Pharmacy, Karad, Dist. Satara, atenolol and amlodipine in tablets by high-performance Maharashtra for providing necessary facilities and thin-layer chromatography. J. Pharm. Biomed. Anal. Shreya Life Sciences Pvt. Ltd. Aurangabad and Lupin 12. Halker U P, Bhandari N P, Rane S H. High performance Research Park, Pune (M. S.) for providing the gift liquid chromatographic simultaneous determination of sample of Amlodipine Besylate and Losartan Potassium amlodipine and enalapril maleate from pharmaceutical preparation. Indian Drugs. 1988; 35: 168. Asian J. Research Chem. 2(1): Jan.-March, 2009 13. Shimooka K, Sawada Y, Tatematsu H. Analysis of 25. Agrekar A P, Powar S G. Simultaneous determination of amlodipine by a sensitive high performance liquid atenolol and amlodipine in tablets by high performance chromatography method with amperometric detection. J. thin layer chromatography. J. Pharm. Biomed. Anal. 14. Yeung P K F, Mosher S J, Pollack PT. High performance 26. Bresford A P, Marcrac P V, Stopher D A. Analysis of liquid chromatography assay for amlodipine: chemical amlodipine in human plasma by gas chromatography. J. stability and pharmacokinetics in rabbits. J. Pharm. 27. Feng Y, Meng Q, Guo X. Human plasma amlodipine GC– 15. Patki R V, Tamhanker C P, Tipnis H P. Simple and rapid MS determination, Guandong Yaoxueyuan Xuebao. 1998; high performance liquid chromatographic estimation of amlodipine in pharmaceutical dosage forms. Indian Drugs 28. Yasuda T, Tanaka M, Iba K. Quantitative determination of amlodipine in serum by liquid chromatography with 16. Josefsson M, Zackrisson A L, Norlander B. Sensitive high atmospheric pressure chemical ionization tandem mass performance liquid chromatographic analysis of spectrometry. J.Mass Spectrom. 1996; 31: 879. amlodipine in human plasma with amperometric detection 29. Mohamed Y E, Naglaa M E K, Bahia A M, Nasshwa G and a single step solid phase sample preparation. J. M. Fluorimetric determination of amiodrone, amlodipine Chromatogr. B; Biomed. Appl. 1995; 672; 310. and propafenone. Bull. Fac. Pharm. 1998; 36: 1. 17. Shang F, Shang K. Determination of amlodipine in tablets 30. Prasad C V N, Parihar C, Chowdhary T R S. by HPLC. ZhoggnoYiyao Gangye Zazhi. 1996; 27: 411. Simultaneous determination of atenolol–amlodipine and 18. European Pharmacopoeia, 3rd ed. Council of Europe, haloperidol– trihexaphenidyl in combined tablet Strasbourg, 2001. pp. 431, (supplement). preparations by derivative spectroscopy. Pharm. 19. Avadhanulu A B, Srinivas J S, Anjaneyulu Y. Reversed phase HPLC determination of amlodipine in drugs and its 31. Prasad C V N, Saha R N, Parimoo P. Simultaneous pharmaceutical dosage forms. Indian Drugs. 1996; 33: 36. determination of amlodipine–enalapril maleate and 20. Sankar S R, Nanjan M J, Vasudevan M. Simultaneous amlodipine–lisinopril in combined tablet preparations by estimation of atenolol and amlodipine in formulations by derivative spectrophotometry. Pharm. Pharmacol. reversed phase-HPLC. Ind J. Pharm. Sci. 1997; 59: 171. 21. Dhorda V J, Shetkar N B. Reversed phase liquid 32. Jain H K, Agrawal R K. Spectrophotometric methods for chromatographic determination of ramipril and simultaneous determination of amlodipine besylate and amlodipine in tablets. Indian Drugs. 1999; 36: 638. lisinopril in tablets. Indian Drugs. 2000; 37: 196. 22. Chandrashekhar T G, Rao P S N , Smrita K. Analysis of 33. Mashru C R, Parikh P P. Development of a method for amlodipine besylate by HPTLC with fluorimetric simultaneous determination of amlodipine besylate and detection: a sensitive method for assay of tablets. J. Planar. Chromatogr. Mod. TLC. 1994; 7: 458. 34. Khopade S A, Jain N K. Difference spectrophotometric 23. Pandya K K, Satia M, Gandhi T P. Detection and determination of amlodipine besylate. Indian drugs. 2000; determination of total amlodipine by high performance thin layer chromatography: A useful technique for 35. Topale P. R. Gaikwad N. J. Tajane M. R. Simultaneous pharmacokinetic studies. J. Chromatogr. B; Biomed. UV- spectrophotometric estimation of losartan potassium and Amlodipine in tablet. Indian drugs. 2003; 40 (2): 119- 24. Ilango K, Kumar P B, Prasad V RV. Simple and rapid determination of amlodipine in pharmaceutical dosage forms. Indian J. Pharm. Sci. 1997; 59: 336.


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Interview from “Fathers and Sons” by Christine Williams. Published by HarperCollins, 1996. Ric Charlesworth, sportsman Born in 1952 on the day King George VI died, Ric Charlesworth grew up in the Perth suburb of Dalkeith which at that time comprised the occasional house, with mostly bushland leading down to the Swan River. He played at swimming, climbing trees, building cubby houses, and m

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