Development and Validation of HPTLC Method for the Simultaneous Estimation of Amlodipine Besylate and Atorvastatin Calcium in Combined Dosage Form
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Department of Pharmaceutical Analysis, College of Pharmacy, India
Department of Pharmaceutical Analysis, A.U college of Pharmacy, Andhra University, India
Publish date: 2017-10-27
Eurasian J Anal Chem 2016;11(3):155–168
This present study reports for the first time simultaneous quantitation of Amlodipine besylate and Atorvastatin calcium by HPTLC from a combined dosage form. Chromatographic separation of the drugs were performed on aluminum plates precoated with silica gel 60 F254 used as stationary phase and the chromatogram was developed using Ethyl acetate: Methanol: Ammonia (7.5 : 2 : 0.5 %v/v/v) as mobile phase. Amlodipine besylate and Atorvastatin calcium showed Rf values 0.50 ± 0.02 and 0.26 ± 0.02 respectively. Densiometric analysis of both the drugs was carried out in the absorbance mode at 365 nm. The method has been successfully applied to tablets and was validated according to ICH Harmonized Tripartite guidelines. The linearity regression analysis for calibration showed 0.9983 (r2) and 0.9994 (r2) for amlodipine besylate and atorvastatin calcium with respect to peak area and height in the concentration range of 100-500ng/spot and 200-600ng/spot respectively. The percentage recovery for amlodipine besylate was found to be 101.82 (at 50%), 99.12 (at 100%) , 100.5(at 150%) and 101.41 (at 50%), 101.71 (at 100%), 99.5(at 150%) for atorvastatin calcium. The limit of detection was 30 ng/spot and 60 ng/spot for amlodipine besylate and atorvastatin calcium respectively. The limit of quantification was found to be 100 ng/spot and 200 ng/spot for amlodipine besylate and atorvastatin calcium respectively. The developed TLC technique is precise, specific and accurate. It was concluded that the developed method offered several advantages such as rapid, cost effective, simple mobile phase and sample preparation steps and improved sensitivity made it specific, reliable and easily reproducible in any quality control set-up providing all the parameters are followed accurately for its intended use.
Budawari S. (2008). The Merck Index, 23rd ed. Whitehouse Station, New Jersey. 516, 6235.
Martindale & Sweetman SC. (1999). The complete drug reference. 32nd ed. Pharmaceutical Press. 822, 907.
Oneil M.J, Smith A, Heckelman P.E & Budawari S. (2001). The Merck Index, an Encyclopedia of Chemicals, Drugs and Biologicals, 13rd ed., Merck and Co Inc., White House Station, New jersey. 488, 865.
Mohammadi A., Rezanour N., Ansari M., Dogaheh, Ghorbani F., Bidkorbeh., Hashem M.& Walker RB. (2007). Stability indicating RP-HPLC assay for simultaneous determination of Atorvastatin and Amlodipine in commercial tablets. Journal of Chromatography B. 846, 215‐221.
Desager J.P. & Hormans Y. (1996). Clinical Pharmacokinetics of 3‐ hydroxy‐3methylglutaryl‐coenzyme A reductase inhibitors. Clin. Pharmacokinet. 31, 348‐371.
Malinowski J.M. (1998). Atorvastatin, A hydroxymethylglutarylcoenzyme A reductase inhibitors. J.Health‐Syst Pharm. 2253‐2267.
Lipitor product information. Park‐Davis/Pfizer Inc. April (2002).
Burnham T.H. (2002). HMG‐CoA reductase inhibitors. In. ed. Drug Facts and Comparisons. Louis Facts and Comparisons, Inc. 536‐542a.
Indian Pharmacopoeia, (2007). Govt. of India, Ministry of Health and Family Welfare, Vol. 2, Delhi. Publication by Controller of Publication. 749-52.
Indian Pharmcopoeia, (2007). Govt. of India, Ministry of Health and Family Welfare, Vol. 2, Delhi. Publication by Controller of Publication.714-16.
British Pharmacopoeia, (2005). International ed. Published on the Recommendation of the Medicines Comissions Pursuant to Medicines Act vol. 1, 138.
The European Pharmacopoeia, (2002). Counsile of Europe, Codex,France, 4th Edn.639-40.
The United States Pharmacopoeia Convention, Inc. (2007). Rockville, M.D. 3496-97, 1532.
Zarghi A., Shafaati A., Foroutan S.M. & Khoddam A., (2005). A simple and rapid HPLC method for the determination of atorvastatin in human plasma with UV detection and its application to pharmacokinetic studies, Arznei mittel forschung. 55, 451-454.
Koytchev R., Ozalp Y., Erenmemisoglu A.,Vander Meer M.J.& Alpan R.S.(2004). Bioequivalence study of atorvastatin tablets, Arzneimittelforschung. 54, 573-577.
Bahrami G., Mohammadi B., Mirzaeei S. & Kiani A. (2005). Determination of atorvastatin in human serum by reversed-phase high-performance liquid chromatography with UV detection, J. Chromatogra. B. 826, 41-45.
Hermann M., Christensen H. & Reubsaet J. L. (2005). Determination of atorvastatin and metabolites in human plasma with solid-phase extraction followed by LC-tandem MS, Anal. Bioanal. Chem. 382, 1242-1249.
Jemal M., Ouyang Z., Chen B. C., & Teitz D. (1999). Quantitation of the acid and lactone forms of atorvastatin and its biotransformation products in human serum by high-performance liquid chromatography with electrospray tandem mass spectrometry, Rapid Commun. Mass Spectrom. 13, 1003-1015.
Bullen W. W. & Miller R. A. (1999). Development and validation of a high-performance liquid chromatography tandem mass spectrometry assay for atorvastatin, ortho-hydroxy atorvastatin, and para-hydroxy atorvastatin in human, dog, and rat plasma, J. Am. Soc. Mass Spectrom. 10, 55-66.
Erturk S., Sevinc A. E., Ersoy L. & Ficicioglu S. (2003). An HPLC method for the determination of atorvastatin and its impurities in bulk drug and tablets, J. Pharm. Biomed. Anal. 33, 1017-1023.
Pasha M. K., Muzeeb S., Basha S. J., Shashikumar D., Mullangi R. & Srinivas N. R. (2006). Analysis of five HMG-CoA reductase inhibitors- atorvastatin, lovastatin, pravastatin, rosuvastatin and simvastatin pharmacological, pharmacokinetic and analytical overview and development of a new method for use in pharmaceutical formulations analysis and in vitro metabolism studies, Biomed. Chromatogr. 20, 282-293.
Manoj K., Shanmugapandiyan P. & Anbazhagan S. (2000). RP-HPLC method for simultaneous estimation of atorvastatin and aspirin from capsule formulation, Indian Drugs. 41, 284-288.
Khopde S.A. & Jain N.K. (2000). Difference spectrophotometric estimation of amlodipine besylate, Indian Drugs. 37, 351-53.
Bahrami G. & Mirzaeei S.(2004). Simple and rapid HPLC method for determination of amlodipine in human serum with fluorescence detection and its use in pharmacokinetic studies, J. Pharm. Biomed. Anal. 36, 163-68.
Streel B., Laine C., Zimmer C., Sibenaler R. & Ceccato A. (2002). Enantiomeric determination of amlodipine in human plasma by liquid chromatography coupled to tandem mass spectrometry, J. Biochem. Biophys. Methods. 54, 357-68.
Carvalho M., Oliveira C. H., Mendes G. D., Sucupira M., Moraes M. E. & De Nucci G. (2001). Amlodipine bioequivalence study: quantification by liquid chromatography coupled to tandem mass spectrometry, Biopharm. Drug. Dispos. 22, 383-90.
Patki R. V., Tamhankar C. P. & Tipnis H. P. (1994). Simple and rapid high performance liquid chromatographic estimation of amlodipine from pharmaceutical dosage, Indian Drugs. 31,560-61.
Avadhanulu A. B., Srinivas J. S. & Anjaneyulu Y. (1996). RP-HPLC determination of amlodipine besylate in drug and its pharmaceutical dosage forms, Indian Drugs. 33, 36-40.
Zarapkar S. S., Katle S. S. & Rane S. H. (1997). HPLC determination of amlodipine and atenolol simultaneously from pharmaceutical preparation, Indian Drugs. 34, 350-53.
Valiyare G. R., Chandra A., Apte S. K. & Mahadik A. A. (2005). HPLC determination of amlodipine, losartan and ramipril in pharmaceutical formulations, Indian Drugs. 42, 309-312.
Kamble N. & Venkatachalam A. (2004). Determination and validation of HPLC method for simultaneous determination of lisinopril and amlodipine from tablet, Indian Drugs. 41, 179-781.
Kulkarni A. P., Gat G. V., Pimple S.V. & Joshi M.A. (2003). HPLC method for determination of losartan potassium and amlodipine besylate in tablets, Indian Drugs. 40, 298-299.
Zarapkar S.S. & Kanyawar N. S. (2002). Simultaneous estimation of amlodipine besylate and losartan potassium in pharmaceutical dosage by RP-HPLC, Indian Drugs. 39, 338-41.
Altiokka G. & Altiokka M. (2002). Flow injection analysis of amlodipine using UV-detection, Pharmazie. 57, 500-503.
Kamat K. & Chaturvedi S. C. (2005). Stability indicating assay method for amlodipine tablets, Indian J.Pharm. Sci. 67, 236-239.
Naidu K. K., Kale U. N. & Shingare M. S. (2005). Stability indicating RP-HPLC method for simultaneous determination of amlodipine and benazepril hydrochloride from their combination drug product. J. Pharm. Biomed. Anal. 39, 147- 155.
Sahu R. & Patel V. B. (2006). Simultaneous spectrophotometric determination of amlodipine besylate and atorvastatin calcium from their binary mixture by dual wavelength and zero absorbance measurement, Indian Drugs. 43, 160-161.
Chaudhari B. G. & Patel N. M. (2006). Development and Validation of HPLC method for simultaneous estimation of Atorvastatin Calcium and Amlodipine Besylate. J. Pharm. Research. 5, 141-144.
Chaudhari B. G., Patel N. M. & Shah P. B. (2007). Stability Indicating RP-HPLC for simultaneous determination of atorvastatin calcium and amlodipine besylate from their combination drug Products, Chem. Pharm. Bull. 55, 241-246.
ICH Q2A (1994). Text on Validation of analytical procedures, International Conference on Harmonization tripartite guidelines. adapted 27 Oct .
Sethi P. D. (1996). High performance thin layer chromatography Quantitative Analysis Pharmaceutical formulations, 1st Edition, CBS Publishers and distributors, India.
Touchstone J .C. (1992). Practice of thin layer chromatography, 3rd Ed., Published by John Wiley and sons, Inc., USA.
Sethi P. D & Charegaonkar D. (1999). Identification of drugs in pharmaceutical formulations by thin layer chromatography, 2nd Ed., CBS Publishers and distributors, USA.
ICH Q2B Text on validation of analytical procedures. (1994). Methodology International Conference on Harmonization. Harmonization tripartite guidelines, adapted 27 Oct.