Determination of (S)-(+)- and (R)-(-)-ibuprofen enantiomers in human plasma after chiral precolumn derivatization by reversed-phase LC–ESI-MS/MS

Abstract

Background: A selective, sensitive and high-throughput LC–ESI-MS/MS method has been developed and validated for the chromatographic separation and quantitation of (S)-(+)-ibuprofen and (R)-(-)-ibuprofen after derivatization with (S)-(-)-1-(1-napthyl)ethylamine using 1-hydroxybenzotriazole as the activator of the carboxylic acid group and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide as the coupling reagent in human plasma. Results: Both the analytes were chromatographically separated with a resolution factor of 1.27 on a Kinetex PFP (50 × 4.6 mm, 2.6 µm) analytical column. The method was validated over the concentration range of 0.10–32.0 µg/ml for both the enantiomers. The magnitude of matrix effect was assessed by post-column analyte infusion and also by precision (%CV) values for the calculated slopes of calibration curves. The mean extraction recovery was >91% for both the enantiomers. Conclusion: The method was successfully applied to a bioequivalence study in 34 healthy human subjects. The assay reproducibility was confirmed by reanalysis of 130 subject samples.