STABILITY INDICATING HPTLC METHOD DEVELOPMENT FOR SIMULTANEOUS ESTIMATION OF ACYCLOVIR AND HYDROCORTISONE IN API AND PHARMACEUTICAL DOSAGE FORM

Abstract

An accurate, precise, sensitive, selective stability indicating High Performance Thin Layer Chromatographic method has been developed and validated for simultaneous estimation of Acyclovir and Hydrocortisone in API and Pharmaceutical dosage form. Chromatographic separation of both the drugs were achieved by using TLC aluminum plates precoated with silica gel 60 F 254 as stationary phase and Toluene: Methanol: Glacial acetic acid in the ratio of 6:4:0.05 v/v/v as mobile phase. Densitometric analysis was carried out in absorbance mode at 250 nm. The Rf value was found to be 0.30 ± 0.02 and 0.73 ± 0.02 for Hydrocortisone and Acyclovir respectively. The method was validated in compliance with ICH guidelines for linearity, Precision, Robustness, Repeatability, Limit of detection, Limit of quantification, Accuracy and Assay. Linear regression analysis data for the calibration curves shows good linear correlation relationship over the concentration ranges from 1000-6000 ng/spot with R² = 0.9997 and 20-1200 ng/spot with R² = 0.9997 for Acyclovir and Hydrocortisone respectively. The Limit of detection and Limit of quantification were 240 and 720 ng/spot for Acyclovir while 400 and 1220 ng/spot for Hydrocortisone. The recoveries of Hydrocortisone and Acyclovir were found to be 98.6 and 99.8%. The drugs were subjected to acidic, basic, neutral, oxidative, thermal and photolytic conditions with below 20% degradation.