Quantitation of PGE9509924, a novel, nonfluorinated quinolone, in rat plasma using liquid chromatography electrospray-tandem mass spectrometry following solid-phase extraction sample clean-up in a 96-well format

PGE9509924, a novel nonfluorinated quinolone, is a potent antibacterial agent with a broad spectrum of activity. A semi-automated method using 96-well format, solid-phase extraction has been developed for quantitating PGE9509924 in rat plasma. The Waters Oasis HLB extraction plate containing a polymeric packing material was found to give the best overall recoveries. All liquid transfer steps other than aliquoting the plasma are accomplished using a 96-channel pipettor. Reverse-phase HPLC with electrospray/MS/MS detection using selective reaction monitoring is used to quantitate the samples. Stable isotopically labeled PGE9509924 is used as the internal standard. The assay is linear over the range from 0.01 to 10 ug/ml. Excellent precision is obtained within a single run and between multiple runs performed on different days. CVs of <6% were observed. The combination of the semi-automated, 96-well parallel sample processing and the short runtime on the LC/MS/MS results in a high throughput assay with reduced operator interaction.