Quantitative Determination of Fexofenadine in Human Plasma by UPLC

Abstract:

A selective, rapid and sensitive reverse phase ultra-performance liquid chromatography method was developed for the quantitative determination of fexofenadine in human plasma. With carbamazepine as internal standard, sample pretreatment involved a one-step extraction with ethyl acetate from 980μl plasma. The sample was analyzed using 10mMKH²PO_⁴ buffer pH 2.5 and acetonitrile (70:30 v/v) as mobile phase. Chromatographic separation was achieved on an ACQUITY UPLC^TM BEH (C-18) column (1.7 μm, 2.1mm x 100mm) using isocratic elution (at a flow rate of 0.25 ml/min). The peak was detected using UV-PDA detector set at 210 nm and the total time for a chromatographic separation was 10 min. Linear calibration curves were obtained in the concentration range of 30.09-1805.39 ng/ml with a lower limit of quantification of 30.09 ng/ml. The inter- and intra- day precision (RSD) values were below 15% and accuracy (RE) was from 1.55 to 5.51 % at all QC levels. The mean recoveries for fexofenadine at high, middle and low quality control samples was obtained 74.3%, 73.2% and 64.8% respectively and for internal standard was 82.8%. Developed method was found to be accurate, precise, selective and rapid for estimation of fexofenadine in plasma and can be used for pharmacokinetic and bioequivalence studies.

Key words: Fexofenadine, UPLC, Human plasma, Liquid-liquid extraction.