Abstract
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. 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. 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.
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