ENANTIOSELECTIVE DETERMINATION OF HYDROXYCHLOROQUINE AND ITS MAJOR METABOLITES IN URINE AND THE OBSERVATION OF A REVERSAL IN THE (+)-(-)-HYDROXYCHLOROQUINE RATIO

A sequential achiral-chiral high-performance liquid chromatographic system has been developed for the quantitation in urine of the enantiomers of hydroxychloroquine (HCQ), and of its 3 major metabolites, desethylhydroxychloroquine (DHCQ), desethylchloroquine (DCQ), and bisdesethylchloroquine (BDCQ). HCQ and its metabolites were separated and quantified on a cyano-bonded phase, and the enantiomeric ratios were determined using a Chiral-AGP chiral stationary phase. The assay validation and application of this method to a preliminary study in a human volunteer are presented. In this subject, the initial 0-4 h urine contained the 2 HCQ enantiomers in a ratio of (+)-HCQ: (-)-HCQ of 3:2; by the 2,064 h of the study, this ratio had reversed to (+)-HCQ: (-)-HCQ of 3:7.