Abstract

DEVELOPMENT AND VALIDATION OF RP-HPLC METHOD FOR ESTIMATION OF ASPIRIN AND TICLOPIDINE HYDROCHLORIDE IN SPIKED HUMAN PLASMA

Wakchaure Varsha S.*, Kachave R. N., Swami Shubhangi M., Tilekar Devika K. and More Archana S.

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Abstract

A sensitive, selective and simple high performance liquid chromatography was developed and validated for the quantification of Aspirin and Ticlopidine hydrochloride in human plasma. Following liquid-liquid extraction, the analyte were extracted from plasma sample by organic solvent like acetonitrile, separated by HPLC on a commercially available column Inertsil C-18 (250 mm × 4.6 mm, 5.0 μm). In a gradient programme mobile phase is of (0.02M) 0.544 gm KH2PO4 in 200ml water (pH-3.9) and ACN( 45:55%v/v) upto 3 min and further change to(35:65%v/v) with 1 ml flow rate. The UV-Vis at λ 254 nm. The correlation coefficient of the caliberation curve was linear (r2 0.99) over the concentration ranged from3.8-6.9 μg ml-1 for aspirin and 0.96-1.73 μg ml-1 for Ticlopidine hydrochloride. The intra-and inter-batch precisions were less than 15% of the relative standard deviation and the accuracy range from to95-103 % in terms of Percent accuracy. The limit of detection of aspirin was 0.38 μg ml- 1and Ticlopidine Hydrochloride was 0.099 μg ml-1. The extraction recovery of both the drug is more than 80%. The proposed method enables the unambiguous identification and quantification of Aspirin and Ticlopidine for pharmacokinetic, bioavailability or bioequivalence studies.

Keywords: Aspirin, Ticlopidine Hydrochloride, RP-HPLC, Method development and validation.