Stereoselective determination of hydroxychloro‐quine and its major metabolites in human urine by solid‐phase microextraction and HPLC

Abstract

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 degrees C and the desorption time was 3 min using methanol (100%). PDMS-DVB 60 microm 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.

Development of a chiral HPLC method for the separation and quantification of hydroxychloroquine enantiomers

“…Thus, we compared two commonly used basic additives, diethylamine (DEA) and triethylamine (TEA). We found that the effects on the resolution by DEA were more obvious (data not shown), which is consistent with previous reports 28,29 . Consequently, we examined the in uence on the resolution by different amounts of DEA.…”

“…To compare biological activities and therapeutic values of HCQ enantiomers, the separation of the enantiomers and the analysis of their optical purity are a prerequisite. Previously, two groups of researchers reported a similar analytical method for the analyses of HCQ enantiomers and their metabolites to investigate the enantiomeric preference on drug metabolism 28,29 . Unfortunately, in addition to the reproducibility issue, the reported resolution of HCQ enantiomers was very poor, which was unable to accurately and quantitatively analyze the actual values of respective enantiomer of HCQ.…”

Enantioselective analysis of mirtazapine, demethylmirtazapine and 8‐hydroxy mirtazapine in human urine after solid‐phase microextraction

“…SPME has been extensively used coupled to GC or GC‐MS for drug analysis with excellent clean‐up and high pre‐concentration capacity 1. SPME has also been used prior to LC or LC‐MS (LC‐MS or LC‐MS‐MS) analysis of a wide variety of compounds, specially thermally unstable, low to nonvolatile or very polar compounds not amenable to GC or GC‐MS analysis 2–12. CE has also been used for the analysis of drugs after SPME extraction 13.…”

Therapeutic drug monitoring by LC–MS–MS with special focus on anti-infective drugs

“…This feature has been used for detection of quinine and metabolites and other antimalarial drugs in human samples, either with classical HPLC [103,104] or with capillary-LC [105]. Several different extraction techniques were used for methods using HPLC with UV detection for the quantification of antimalarial drugs: liquid-phase microextraction [106], solid-phase microextraction [107], automated solid-phase extraction [108], and extraction with restricted-access materials [109]. As sample material, many methods use dried blood spots, either with UV detection [110][111][112][113], or fluorescence detection [114].…”