Rapid determination of gemifloxacin in human plasma by high-performance liquid chromatography-tandem mass spectrometry

A method was developed for the determination of gemifloxacin (I) in human plasma using high-performance liquid chromatography-tandem mass spectrometry. Prior to analysis, the protein in plasma samples was precipitated with acetonitrile containing [(CH3)-C-13-H-2] gemifloxacin (II) to act as an internal standard. The supernatant was injected onto a PLRP-S column without any further clean-up. The mass spectrometer was operated in positive ion mode, employing a heat assisted nebulisation, electrospray interface. Ions were detected in multiple reaction monitoring (MRM) mode. The assay requires 50 mu l of plasma and is precise and accurate within the range 10-5000 ng/ml. The average within-run and between-run coefficients of variation were <11% at 10 ng/ml and greater concentrations. The average accuracy of validation standards was generally within +/-7% of the nominal concentration. There was no evidence of instability of I in human plasma following three complete freeze-thaw cycles and samples can safely be stored for at least 6 months at -20 degrees C. The method proved very robust and was successfully applied to the analysis of clinical samples from patients dosed with gemifloxacin. (C) 2000 Elsevier Science B.V. All rights reserved.