An HPLC method with diode array detector for the simultaneous quantification of chloroquine and desethylchloroquine in plasma and whole blood samples from Plasmodium vivax patients in Vietnam, using quinine as an internal standard

Abstract

© 2016 The Authors Biomedical Chromatography Published by John Wiley & Sons Ltd. A sensitive, simple method for quantification of chloroquine (CQ) and desethylchloroquine (MCQ) in whole blood and plasma from Plasmodium vivax patients has been developed using HPLC with diode array detection (DAD). Solid-phase extraction on Isolute-96-CBA was employed to process 100μL of plasma/whole blood samples. CQ, MCQ and quinine were separated using a mobile phase of phosphate buffer 25mm, pH2.60-acetonitrile (88:12, v/v) with 2mm sodium perchlorate on a Zorbax SB-CN 150×4.6mm, 5μm column at a flow rate of 1.2mL/min, at ambient temperature in 10min, with the DAD wavelength of 343nm. The method was linear over the range of 10-5000ng/mL for both CQ and MCQ in plasma and whole blood. The limit of detection was 4ng/mL and limit of quantification was 10ng/mL in both plasma and blood for CQ and MCQ. The intra-, inter- and total assay precision were <10% for CQ and MCQ in plasma and whole blood. In plasma, the accuracies varied between 101 and 103%, whereas in whole blood, the accuracies ranged from 97.0 to 102% for CQ and MCQ. The method is an ideal technique with simple facilities and instruments, bringing about good separation in comparison with previous methods.