Method Development and Validation by RP-HPLC for Estimation of Acyclovir and Hydrocortisone in Bulk and Pharmaceutical Dosage Form

Abstract

A rapid and reliable Reverse Phase High-Performance Liquid Chromatography (RP-HPLC) method was developed and validated for the simultaneous estimation of Acyclovir and Hydrocortisone in bulk and pharmaceutical dosage forms. Chromatographic separation was achieved using a Waters HPLC system equipped with an auto sampler and PDA detector (Model 996), employing a Gemini C18 column (4.6 × 150 mm, 5 µm) maintained at 30°C. The mobile phase consisted of Acetonitrile and Water in a 50:50 v/v ratio, with a flow rate of 1.0 mL/min. Detection was carried out at 238 nm, with an injection volume of 10 µL and a total run time of 8 minutes. The method was validated as per ICH guidelines, demonstrating excellent linearity, precision, accuracy, specificity, and robustness. The retention times of both drugs were distinct and interference-free. This method is efficient, economical, and well-suited for routine quality control and analysis of Acyclovir and Hydrocortisone in pharmaceutical preparations.