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Determination of ketorolac enantiomers in plasma using enantioselective liquid chromatography on an α1-acid glycoprotein chiral stationary phase and ultraviolet detection

Journal of Chromatography B Biomedical Sciences and Applications
Publication Date
DOI: 10.1016/0378-4347(94)00228-2
  • Biology
  • Pharmacology


Abstract A chirally selective high-performance liquid chromatographic assay was developed to measure the R(+) and S(−) enantiomers of ketorolac in plasma for pharmacokinetic studies. Naproxen sodium [ S(+) enantiomer] (10 μg) was used as an internal standard. Plasma samples (0.5 ml) were acidified (50 μl of 4 M H 3PO 4 to pH 1.5), extracted into 0.4 ml of 10% pentan-2-ol in hexane and back-extracted into 0.15 ml of base (20m M NaOH pH to 7–8), of which samples (5 μl) were chromatographed on a 100 × 4 mm I.D. column packed with an HPLC chiral stationary phase based upon immobilized α 1-acid glycoprotein (Chiral AGP-CSP) with 4% propan-2-ol in 0.1 M NaH 2PO 4 pH 5.5, at 0.9 ml/min. Detection was at 325 nm and run time was 10 min. Retention times of R- and S- ketorolac and of S(+)-naproxen were 3.3, 4.8 and 6.4 min, respectively. The metabolite p-hydroxyketorolac was not resolved enantiomerically and had a retention time of 2.2 min. The assay was linear over the range 0.5–10 mg/l, with precisions <5% C.V. Good separations (α > 1.35) and resolutions ( R g > 3.23) between peaks were achieved. The sensitivity could be extended to 35 μg/l with less precision by increasing the injection volume to 100 μl.

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