Development of a chiral assay for a novel, nonfluorinated quinolone, PGE-9509924, in dog plasma using high performance liquid chromatography with electrospray tandem mass spectrometry or fluorescence detection.
PGE-9509924 is a nonfluorinated quinolone and is active against a variety of susceptible and drug resistant bacteria in vitro and in animal infection models. A method for determining both enantiomers of PGE-9509924 in dog plasma has been developed. The enantiomers are derivatized with a chiral derivatizing agent, (-)-1-(9-fluorenyl)ethyl chloroformate (FLEC) and the resulting diastereomers are separated by reverse phase chromatography. Plasma samples are prepared via solid phase extraction (SPE) in a 96-well format prior to being derivatized. Samples are then analyzed by electrospray-LC/MS/MS with multiple reaction monitoring or by HPLC with fluorescence detection. Results of a side-by-side validation of the method with LC/MS/MS and HPLC/Fl detection are presented. Over the range selected for validation (0.025-10 micro g/ml), both methods give similar results with identical limits of quantitation. Due to the selectivity of LC/MS/MS and the use of a stable-isotopically labeled internal standard, significantly shorter chromatographic runtimes are achieved with LC/MS/MS, making it the method of choice for sample analysis.
Zoutendam
P H
PH
Health Care Research Center, Procter and Gamble Pharmaceuticals Inc, 8700 Mason-Montgomery Road, Mason, OH 45040, USA. zoutendam.ph@pg.com
Canty
J F
JF
Martin
M J
MJ
Dirr
M K
MK
D000818:Animals; D000890:Anti-Infective Agents / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D004285:Dogs; D007202:Indicators and Reagents; D015363:Quinolones / Q000097:blood*; D012015:Reference Standards; D015203:Reproducibility of Results; D013050:Spectrometry, Fluorescence; D021241:Spectrometry, Mass, Electrospray Ionization; D013237:Stereoisomerism
D000890:Anti-Infective Agents; D007202:Indicators and Reagents; C466659:PGE 9509924; D015363:Quinolones