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Stereoselective determination of hydroxychloro‐quine and its major metabolites in human urine by solid‐phase microextraction and HPLC
Author(s) -
Moraes de Oliveira Anderson Rodrigo,
Sueli Bonato Pierina
Publication year - 2007
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.200700121
Subject(s) - chromatography , chemistry , high performance liquid chromatography , urine , stereoselectivity , enantiomer , metabolite , solid phase extraction , extraction (chemistry) , diethylamine , stereochemistry , organic chemistry , biochemistry , catalysis
The enantioselective analysis of hydroxychloroquine (HCQ) and its major metabolites was achieved by HPLC and solid‐phase microextraction. The chromatographic separation was performed on a Chiralcel OD‐H column using hexane/methanol/ethanol (96:2:2, v/v/v) plus 0.2% diethylamine as the mobile phase, at the flow rate of 1.3 mL/min. The main extraction parameters were optimized. The best condition was achieved by the addition of 10% NaCl and 1 mL phosphate buffer 1 mol/L pH 11 to 3 mL human urine. The extraction was conducted for 40 min at 25°C and the desorption time was 3 min using methanol (100%). PDMS‐DVB 60 μm fiber was used in this study. The mean recoveries were 9.3, 9.2, and 14.4% for HCQ, desethylhydroxychloroquine (DHCQ), and desethylchloroquine (DCQ), respectively. The method was linear over the range of 50–1000 ng/mL for HCQ enantiomers and over the range of 42–416 ng/mL for DCQ and DHCQ enantiomers. Within‐day and between‐day precision and accuracy assays for HCQ and its metabolites were lower than 15%. The preliminary 48 h urinary excretion study performed in human urine showed to be stereoselective. The amount of (+)‐( S )‐enantiomer excreted was higher than its antipode.