Analysis of ketoprofen enantiomers in human and rat plasma by hollow-fiber-based liquid-phase microextraction and chiral mobile-phase additive HPLC

A simple, inexpensive, and efficient preconcentration and cleanup three-phase hollow fiber liquid-phase microextraction method (HF-LPME) was developed for the extraction of the anti-inflammatory ketoprofen (KTP) from human and rat plasma and HPLC enantioseparation of its enantiomers using vancomycin as a chiral mobile-phase additive with an achiral C-8 column. The effects of different parameters influencing the efficiency of extraction were optimized for aqueous samples. Under optimized conditions, KTP enantiomers were extracted from 0.5 mL of plasma diluted to 5 mL with salinated and acidified deionized water (pH = 2) with 25 mu L of alkalinized acceptor phase (pH = 11) during 30 min at room temperature. The mean recoveries of (-)-(R)-and (+)-(S)-KTP were 72.8% and 70.9%, respectively. The quantification limit was 20 ng/mL with linear response over the 20-2000 ng/mL concentration range for both enantiomers. Assay precision was studied within-day and between-day using 100 ng/mL KTP solutions. For both KTP enantiomers, relative standard deviations were lower than 12%. The proposed microextraction method was applied for the extraction of KTP enantiomers from human and rat plasma samples after oral administration of pure (+/-)-KTP in 4 mg/kg dosage for rats and 50 mg dosage for humans to assess the enantiospecific bioavailability of KTP enantiomers in their plasma. In vivo inversion studies revealed that the bioavailability of S-KTP is higher than that of the R enantiomers in rat, but they are similar in human plasma. The developed method showed that HF-LPME is a promising technique for sample preparation for the analyses of chiral drugs in biological samples.