Asian J. Research Chem.. 2(1): Jan.-Mar. 2009 ,
www.ajrconline.org
ISSN 0974-4169 RESEARCH ARTICLE
Development and Validation of Method for Simultaneous Estimation of Atorvastatin Calcium and Ramipril from Capsule Dosage Form by First Order Derivative Spectroscopy. Thamake SL, Jadhav SD, Pishawikar SA*
Department of Pharmaceutical Chemistry, Bharati Vidyapeeth College of Pharmacy, Near Chitranagari, Kolhapur-416013 * Corresponding Author E-mail:
[email protected]
ABSTRACT
Atorvastatin Calcium and Ramipril are used in combination for treatment of hypertension. The present work deals with simple spectrophotometric method development for simultaneous estimation of Atorvastatin Calcium (ATR) and Ramipril (RAM) in two component capsule formulation. The method employed first order derivative spectroscopy1. For determination of sampling wavelength 10 µg/ml of each of ATR and RAM were scanned in 200-350 nm range and sampling wavelengths were 294 nm for ATR were RAM showed zero crossing point and 229 nm for RAM were ATR showed zero crossing point in first order derivative spectroscopy. For this method linearity observed in 5-40 µg/ml for ATR and 2-20 µg/ml for RAM. The recovery studies confirmed accuracy of proposed method and low values of standard deviation confirmed precision of method. The method is validated as per ICH guidelines.
KEY WORDS
Atorvastatin Calcium, Ramipril, Derivative spectroscopy
INTRODUCTION:
Atorvastatin Calcium is chemically described as [R-(R*, R*)]-2-(4-fluorophenyl)- , dihydroxy-5-(1methylethyl)-3-phenyl-4-[(phenylamino) carbonyl]-1Hpyrrole-1-heptanoic acid calcium salt trihydrate is an anti hyperlipoproteinemic agent act by inhibiting HMG-CoA reductase2-3 and Ramipril is 2S-[1[R*(R*)], 2 , 3 , 6 ]]-1-[2[[1-(Ethoxycarbonyl)-3-phenylpropyl]amino]1-oxopropyl] octahydrocyclopenta [b] pyrrole-2carboxylic acid. Used as ACE inhibitor.2-3 Many analytical methods like HPLC4, HPTLC, electrochemical, radioimmunoassay were reported for determination of ATR alone and combination with other antihypertensive drugs and HPLC method reported for simultaneous estimation of ATR and RAM. However no spectrophotometric method is reported till date for simultaneous determination of these drugs. In this communication we report a new UV spectrophotometric method using derivative spectroscopy.
Reagents and chemicals: Atorvastatin Calcium and Ramipril supplied by Cipla Ltd. India. All solvents were spectrophotometric grade obtained from SD fine chemicals. Water purified by glass distillation apparatus. Method: Stock solutions were prepared separately in water: methanol (60:40) to obtain 100 µg/ml of all drugs. The nine working mixed standard were prepared by dilution of stock solution in same solvent system in concentration range 5-40 µg/ml of ATR and 2-20 µg/ml for RAM. Atorvastatin Calcium and Ramipril initially canned for determining sampling wavelength in range 200-300 nm. Sampling wavelengths were 294 nm for ATR where RAM showed zero crossing point and 229 nm for RAM where ATR showed zero crossing point. Calibration graphs were constructed from the absorbances at respective wavelength.
MATERIALS AND METHODS:
Instrument: Spectrophotometric analysis was carried out on a JASCO UV-spectrophotometer 530 using a 1 cm quartz cell. The instrument settings were zero order and first derivative mode and band width of 2.0 nm in the range of 200–350 nm. Received on 05.01.2009 Accepted on 25.02.2009
Modified on 11.02.2009 © AJRC All right reserved
Asian J. Research Chem. 2(1): Jan.-March, 2009;Page 52-53
Fig. 1: Overlain spectra of ATR and RAM in first order derivative mode.
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Asian J. Research Chem.. 2(1): Jan.-Mar. 2009 ,
Analysis of commercial formulation: Content of twenty capsules were taken and powder equivalent to 10 mg of ATR taken and added in 60 ml of solvent system sonicated for 10 min after sonication volume was made up to 100 ml. 1ml of this stock solution was diluted to 10 ml to get concentration equal to 10 µg/ml of ATR and 5 µg/ml of RAM. This solution is scanned in range 200-350 nm taking solvent system as blank. The spectra obtained was converted first order derivative spectra absorbances were noted and concentrations were determined from regression equations generated from calibration graph.
RESULTS AND DISCUSSION:
Sampling wavelengths were determined from scanning individual drug samples in 200-350 nm range. Sampling wavelengths were 229 nm and 294 nm for RAM and ATR respectively in first order derivative mode. For this method equations generated were Y=-0.0002+0.0006x (r2=0.9999) and Y=-0.0004+0.0008x (0.9999) for ATR and RAM respectively. Linearity of proposed method was found to be 2-20 µg/ml for RAM and 5-40 µg/ml for ATR. Limits of detection were found to be 0.0147 and 0.056 µg/ml of ATR and RAM respectively. Limits of quantitation were found to be 0.041 and 0.18 µg/ml for ATR and RAM respectively. Results of capsule analysis were reported in table 1, result of precision studies and recovery study reported in table 2 and 3 respectively. TABLE 1: RESULTS OF COMMERCIAL FORMULATION
ANALYSIS
OF
Analyte
Label % Label claim RSD* claim estimated (Mean (mg/tab) ± SD*) ATR 10 100.22±0.8722 0.8702 RAM 5 100.10±0.8064 0.8055 SD: Standard Deviation, RSD: Relative Standard Deviation. *Denotes average of six determinations. TABLE 2: RESULTS OF PRECISION STUDIES Analyte Label claim Amount % label (mg/tab.) estimated claim ±SD* ATR 10 10.112 101.12 ± 0.7806 RAM 5 5.005 100.10 ± 0.7402 SD: Standard Deviation. *Denotes average of six determinations.
ACKNOWLEDGEMENTS:
Authors are thankful to Cipla Ltd. India, for providing the free gift samples of Atorvastatin Calcium and Ramipril authors are also thankful to Principal, Bharati Vidyapeeth College of Pharmacy, Kolhapur for providing necessary facilities for this work. TABLE 3: RESULTS OF RECOVERY STUDIES Analyte % Recovery estimated RSD* (Mean ± SD*) ATR 99.10±0.7554 0.7622 RAM 100.15±0.8743 0.8729 SD: Standard Deviation, RSD: Relative Standard Deviation. *Denotes average of six determinations.
REFERENCES: 1.
Beckett AH, Stenlake JB. Practical Pharmaceutical Chemistry. Part 2, CBS Publishers and Distributors. 2002; 4th ed: p. 282-296. 2. Indian Pharmacopoeia, Vol. II, Government of India, The Controller of Publication, New Delhi. 2007; p. 749. 3. Budawari S. The Merck Index, Merck and Co., Inc., Whitehouse Station, NJ. 1996; 23rd ed: p. 893, 8280. 4. Lucie N, et al. HPLC methods for the determination of simvastatin and atorvastatin. TrAC Trends in Anal Chemistry. 2008; 27: 352-367. 5. Ramakrishna N, et al. Chromatography–mass spectrometry methods for the quantitation of statins in biological samples. J Pharma and Biomed Anal. 2007; 44 (2): 379-387. 6. Gholamreza B, et al. Determination of atorvastatin in human serum by reversed-phase high-performance liquid chromatography with UV detection. J Chromatogr B. 2005; 826(1): 41-45. 7. Sahu R, Patel VB. Simultaneous spectrophotometric determination of Amlodipine Besylate and Atorvastatin Calcium in Binary Mixtures. Ind J Pharma Sci. 2007; 69 (1): 110-111. 8. Shaha DA, et al. Development and Validation of a RP-HPLC Method for Determination of Atorvastatin Calcium and Aspirin in Capsule Dosage Form. Ind J Pharma Sci. 2007; 69 (4): 546-549. 9. Shaha DA, et al. RP-HPLC Method for Determination of Atorvastatin calcium and Nicotinic acid and in Combined tablet dosage form. Ind J Pharma Sci. 2007; 69 (5): 700-703. 10. Manoj K, et al. RP-HPLC Method for simultaneous estimation of Atorvastatin and Aspirin from capsule formulation. Indian Drugs. 2004; 41 (5): 284-289.
CONCLUSION:
The method used is simple and rapid and does not involve the use of complex instrument, low value of standard deviation showed that the method is precise and high percentage of recovery of as shown in table shows that the method is accurate.
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