DETERMINATION OF DRUG CODE 456/9 BY DIRECT ESTIMATION METHOD IN HUMAN PLASMA AND TABLET DOSAGE FORM BY RP-HPLC

A simple, precise, rapid and high through put, direct estimation Reverse Phase High Performance Liquid Chromatography (RP-HPLC) method has been developed and validated for quantitation of code 456/9 in human plasma and tablet dosage form. Direct estimation method involves the precipitation of plasma proteins with organic solvent. Clear supernatant was then injected in to the column. The chromatographic separation was performed using methanol: water (80:20, v/v) as isocratic mobile phase at a flow rate of 0.5ml min-1 and InertsilODS-3 (C-18) Column (5 µm, 250mm x 4.60mm) as stationary phase. Detection was carried out using a UV PDA detector at 235 nm. The retention time for code 456/9 was 3.8 ± 0.2. The method was validated for a working range of 60-140ng/ml range and 600-1400ng/ml for plasma and tablet dosage form respectively. The average recovery of extraction procedure was found to be 76%. The limit of detection was found to be 7.7 ± 0.2 ng/ml for plasma. The R.S.D for both intra and inter day precision was found to be less than 10% for both plasma and tablet dosage form. The accuracy for tablet dosage form was found to be in the range of 99.0-100.1%.