Abstract

VALIDATED RP-HPLC/UV METHOD FOR THE QUANTITATION OF DIPHENHYDRAMINE IN FORMULATION VEHICLE AND ITS APPLICATION TO HOMOGENECITY AND STABILITY IN DOSE FORMULATION VEHICLE

Vinay Dhiman* and N. B. Sridhara Murthy

ABSTRACT

A novel, simple, specific, sensitive and reproducible high-performance liquid chromatography (HPLC) assay method has been developed and validated for the estimation of diphenhydramine (DPH). The HPLC method for the analysis of diphenhydramine hydrochloride in dose formulation samples using Tolbutamide as internal standard (IS). The chromatographic analysis was performed by Waters Alliance High Performance Liquid Chromatographic system equipped with UV and PC based data system with Empower Software. Chromatographic separation was achieved using Symmetry sheild, RP 18, 5 μM, (Waters) column maintained at 35ºC temperature and an gradient mobile phase (0.01 M Ammonium Acetate in Milli- Q Water, pH adjusted to 5 using Acetic Acid and acetonirile) at a flow rate of 1 mL/min with a total run time of 10 min. The elute was monitored using an UV detector set at 267 nm. Method of validation was performed as per ICH guidelines and the results met with the acceptance criteria. The calibration curve was linear over a concentration range of 1001 to 14987 μg/mL (r2 = 0.999). Precision & accuracy batches were analysed at 3 different occasions revealed the accuracy of DPH in formulation was in the range of 99.85 to 105.83%, 100.19 to 101.81% and 94.94 to 95.02%, respectively. (with in batches ): Inter day accuracy was 99.8, 98.3 and 98.1 respectively.. The validated HPLC method was successfully applied for formulation analysis.

Keywords: Diphenhydramine, HPLC method validation, Homogeneity, Formulation.