Method Development and Validation for the Simultaneous Analysis of Miconazole Nitrate, Hydrocortisone and Chlorocresol in a Pharmaceutical Topical Cream by Reverse Phase Liquid Chromatography

Abstract

Miconazole/Hydrocortisone (HCA) cream is a pharmaceutical formulation developed as a combination of three major components (HCA, miconazole nitrate [MCN] and chlorocresol [CHL]). In some instances, reference monographs used for referencing the quantification of pharmaceutical molecules focus on individual analysis of molecules. Therefore, there is need to develop, optimize and validate methods that simultaneously separate constituents of a combined formulation. In this study, a method for the separation of HCA, MCN and CHL in miconazole/HCA cream was developed by investigating the effect of mobile phase, stationary phase, wavelength and gradient program on the separation of HCA, MCN and CHL in miconazole/HCA cream. A 10 mM ammonium acetate (AAC) buffer pH 5.0 was used. Acetonitrile was used as the organic modifier. An injection volume of 10 μL was used on a C18, 15 cm × 4.6 mm Zorbax. The optimum conditions for reverse phase liquid chromatography were obtained as follows: injection volume (10 μL), column temperature (30◦C), flow rate (1 mL/min), gradient elution program: 0 min, 60AAC:40ACN; 4 min, 60AAC:40ACN; 9 min, 5AAC:95ACN; 13 min, 5AAC:95ACN using a C18 stationary phase, UV detector, 5 μm stationary phase pore size and post run of 3 min