Analytical Method Development and Validation of Atorvastatin Calcium and Vitamin D3 in Pharmaceutical Dosage Form

Abstract

Purpose: To develop and validate a novel reverse phase high performance liquid chromatographic (RP-HPLC) method for the estimation of atorvastatin and vitamin D3 in pharmaceutical dosage form. The RPHPLC Method has shown adequate separation of Atorvastatin Calcium and Vitamin D3 in Bulk and its synthetic mixture. Method: Chromatographic identification was achieved on Phenomenex C18 (250mm x 4.6mm 5μm particle size) column with mobile phase using Acetonitrile: Methanol ratio of 75:25 v/v, pH 3.5 adjusted with orthophosphoric acid and flow rate is 1.2 ml/min. Sample injection volume is 20 μl and detection wavelength at 252 nm. The developed HPLC method was validated according to International Conference on Harmonization (ICH) Q2 (R1) guidelines. Result: The retention time for Atorvastatin calcium and Vitamin D3 was obtained as 2.9 min and 16.6 min respectively. The linearity of the proposed method was investigated in the range of 8-28 μg/ml and 2-7 μg/ml for Atorvastatin calcium and Vitamin D3 respectively. Correlation coefficient was 0.9984 and 0.9955 for Atorvastatin Calcium and Vitamin D3, respectively. The developed method was validated as per ICH guidelines, for its accuracy, precision, LOD & LOQ. CONCLUSION: The method was developed successfully and the results of developed method were found satisfactory, thus the method is specific, rapid, simple and sensitive for estimation of Atorvastatin Calcium and Vitamin D3.