Abstract

ANALYTICAL METHOD DEVELOPMENT AND VALIDATION FOR GEMICITABINE HCl IN BULK AND PHARMACEUTICAL DOSAGE FORM BY RP-HPLC METHOD

B. Siddartha*, Dr. I. Sudheer Babu, Ch. Ravichandra Gupta, C. Parthiban

Abstract

A simple, sensitive and rapid reverse phase high performance liquid chromatographic method was developed for the estimation of gemicitabine hydrochloride in pure and pharmaceutical dosage forms. A Hypersil BDS C18 column (250 x 4.6mm x 5 μ) was used as a stationary phase with a mobile phase containing a mixture of buffer and acetonitrile in the ratio of 93:7v/v. The flow rate was 1.0ml/min, effluent was monitored at 275nm and eluted at 3.927min. Calibration curve was plotted with a range from 10-60μg/ml for gemicitabine hydrochloride and the correlation was found to be 0.9997. The accuracy range was found between 99.20% and 101.24%. The % RSD values for both intraday and interday precision were less than 1%. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 0.296μg/ml and 0.896μg/ml respectively. The assay was validated for the parameters like specificity, system suitability, precision, accuracy, robustness and ruggedness parameters. The proposed method will be useful for the routine determination of gemicitabine hydrochloride in pharmaceutical dosage form

Keywords: Gemicitabine HCl, Calibration curve, RP-HPLC, Validation.