Development and validation of an eco-friendly HPLC-UV method for determination of atorvastatin and vitamin D3 in pure form and pharmaceutical formulation

Statin-associated muscle symptoms are considered as obvious adverse effects of prolonged statin therapy such as myopathy, myalgia, and rhabdomyolysis. These side effects are associated with vitamin D-3 deficiency and can be adjusted by amendment of serum vitamin D-3 level. Green chemistry aims to decrease the harmful effects of analytical procedures. Here we have developed a green and eco-friendly HPLC method for the determination of atorvastatin calcium and vitamin D-3. The two drugs were separated in less than 10 min on Symmetry column C-18 (100 x 4.6 mm, 3.5 mu m) using a mixture consisting of 0.1% ortho-phosphoric acid (OPA) (pH = 2.16) and ethanol as the mobile phase in gradient manner. We have used Green Analytical Procedure Index (GAPI) tools and the Analytical GREEnness Metric Approach (AGREE) for assessment of the greenness of our proposed method. The method proved linearity over concentration ranges of (5-40) and (1-8) mu g/ml with low limit of detection of 0.475 and 0.041 mu g/ml for atorvastatin calcium and vitamin D-3 respectively. The method was successfully validated in accordance with ICH instructions and utilized for determination of the drugs of interest either in pure form or in their pharmaceuticals.