Abstract

BIOANALYTICAL METHOD DEVELOPMENT, VALIDATION AND QUANTIFICATION OF BOSENTAN BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY IN RAT PLASMA

Nallakumar P.* and Siva Kumar R.

Abstract

A Simple and rapid bioanalytical high performance liquid chromatographic (HPLC) method for the determination of Bosentan using Losertan as an internal standard was developed and validated as per regulatory requirements. Sample preparation was accomplished through liquid phase extraction and chromatographic separation on a reverse phase column. The mobile phase consists of mixture of methanol and water in the ratio of 50:50 at a flow rate of 1ml/min. The wavelength used for the detection of bosentan was 225nm with a total run time of 6minutes. The retention times of bosentan and losertan were found to be 2 and 4 respectively. The method was developed and tested for the linearity range of 250-750ng/ml. The method was validated for accuracy, precision, linearity and recovery in compliance to international regulatory guidelines.

Keywords: Bosten, HPLC, Validation.