Development and Validation of a RP-Ultra performance liquid chromatographic Method for Quantification of Topotecan
P K Saini1, C L Jain, R M Singh
1Research and Development Division, Indian Pharmacopoeia Commission, Sector-23, Rajnagar Ghaziabad-201 001, India.
Abstract:
A simple, very fast, precise and accurate reverse phase ultra performance liquid chromatographic method was developed for the determination and validation of topotecan hydrochloride in bulk and injection dosage form. A Waters BEH C18, 50×2.1 mm, 1.7 μm particle size column in gradient mode was used with mobile phase comprising of 0.1% v/v orthophosphoric acid in water and acetonitrile. The analytical column was thermostated at 50° and flow rate was set at 0.4 ml per min, with photo diode array detection at 260 nm. The retention time of topotecan was found 1.38 min. The method was validated in terms of linearity, accuracy, precision and specificity. The calibration curve was found linear between 20 to 60 μg/ml. The limit of detection and limit of quantification were found 0.2353 and 0.7131 μg/ml, respectively. Percentage recoveries were obtained in the range of 98.91% and 99.17%. The proposed method is precise, accurate, selective and reproducible. The ultra performance liquid chromatographic assay procedure, which proved superior because of its greater sensitivity and relatively shorter (4 min) run time, should be an important tool for speedy future analysis of topotecan hydrochloride in bulk and its injection dosage form.
