Determination of hydroxychloroquine and its major metabolites in plasma using sequential achiral-chiral high-performance liquid chromatography.

A sequential achiral-chiral high-performance liquid chromatographic system has been developed for the determination of the enantiomers of hydroxychloroquine, (+)-HCQ and (-)-HCQ, and the enantiomers of its three major metabolites, bisdesethylchloroquine, desethylhydroxychloroquine and desethylchloroquine, in plasma. The HCQ was separated from the metabolites and interfering components in the plasma and quantified on a cyano-bonded phase, and the enantiomeric composition determined using a Chiral-AGP chiral stationary phase. The assay was validated and applied to the analysis of a pilot study of the pharmacokinetics of (+)- and (-)-HCQ in rabbits.