Liquid chromatography-tandem mass spectrometric determination of a new antibacterial agent (AVE6971) in human white blood cells.
An LC-MS/MS assay for the quantitative determination of a new antibacterial agent (AVE6971) has been developed and validated in human white blood cells (WBC). The assay involved a lysing procedure of white blood cells and ultra centrifugation of the extracts. Chromatography was performed on a Supelcosil ABZ+ C(18) (2.1 mm x 50 mm, 5 microm) column using a mobile phase consisting of methanol/acetonitrile/10mM ammonium formate mixture (10:30:60, v/v/v) at a flow rate of 0.2 ml/min. The linearity was within the range of 10-10000 ng/ml of extracts, corresponding to 0.5-500 ng of AVE6971 in WBC pellets tubes. The validated lower limit of quantification was 10 ng/ml. The inter- and intra-run coefficients of variation (CV) for the assay were <12.9% and the accuracy were from -9.0 to -1.2%. AVE6971 was stable in WBC for at least 1 month at -75 degrees C. This assay proved to be suitable for the determination of AVE6971 in WBC from clinical studies.
Salvador
Arnaud
A
CEPHAC Europe, 90 Avenue des Hauts de la Chaume, BP 28, 86281 Saint-Benoît, France. arnaud.salvador@univ-lyon1.fr
Gautier
Jean-Yves
JY
Pasquier
Olivier
O
Merdjan
Henri
H
D000900:Anti-Bacterial Agents / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D006801:Humans; D007962:Leukocytes / Q000737:chemistry*; D011804:Quinolines / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D015203:Reproducibility of Results; D021241:Spectrometry, Mass, Electrospray Ionization; D053719:Tandem Mass Spectrometry / Q000379:methods*
C524562:AVE6971; D000900:Anti-Bacterial Agents; D011804:Quinolines