DETERMINATION OF TRIFLUSAL IN HUMAN PLASMA BY HIGH PERFORMANCE LIQUID CHROMATOGRAPHY WITH AUTOMATED COLUMN SWITCHING SYSTEM

Abstract

An analytical method of triflusal in human plasma was developed using semi-microbore HPLC equipped with automated column switching system. p-Toluic acid which is the structural analogue of triflusal, was used as an internal standard and 2 M HCl was employed as a stabilizer. The load phase and mobile phase were prepared using acetonitrile and 20 mM KH2PO4 with the volume ratios of 10:90 (pH 2.5) and 43:57 (pH 2.3), respectively. The signals were monitored by a UV detector at 275 nm with flow-rate of load phase, 500 μL/min, and mobile phase, 100 μL/min, respectively. The retention time of triflusal and p-toluic acid was about 20.2 min and 16.4 min, respectively. The detection limit of triflusal in human plasma was 0.01 μg/mL and the limit of quantitative analysis was 0.05 μg/mL. The accuracy of the assay was from 97.76% to 116.51% while the intra-day and inter-day coefficient of variation of the same concentration range was less than 15%. This analytical method demonstrated excellent sensitivity, reproducibility, specificity, and speed using the plasma sample. This method could be successfully applied to evaluate the bioavailability of triflusal in human subjects without time-consuming sample clean-up after oral administration of low dose.