SIMULTANEOUS DETERMINATION OF CHLOROQUINE AND QUININE IN HUMAN BIOLOGICAL-FLUIDS BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

A high-performance liquid chromatographic method with fluorescence detection is described for the simultaneous measurement of quinine, chloroquine and mono- and bidesethylchloroquine in human plasma, erythrocytes and urine. After a liquid-solid extraction on a Bond Elut C-8 cartridge, the compounds are separated on an Inertsil silica column by gradient elution; the mobile phase is a mixture of acetonitrile and methanol-25% ammonia solution (92.7:7.5, v/v). The eluent was monitored with a fluorescence detector (excitation wavelength 325 nm and emission wavelength 375 nm). The limit of detection was ca. 5 ng/ml for chloroquine and ca. 23 ng/ml for quinine. No chromatographic interferences could be detected from endogenous compounds or other antimalarial drugs. The method is accurate with inter- and intra-assay coefficients of variation lower than 7%. Hydroxychloroquine is used as an internal standard because of its structural similarity to chloroquine. The procedure requires 30 min and can be used for therapeutic drug monitoring.