leukotriene-b4 and pentafluorobenzyl-bromide

We have developed a highly sensitive and highly selective method for the determination of Leukotriene B4 (LTB4) in human plasma using negative ion chemical ionization/gas chromatography/tandem mass spectrometry (NICI/GS/MS/MS) analysis. The developed method was summarized as follows. Deuterated LTB4 (d4-LTB4) was added to human plasma samples as an internal standard, and samples were extracted by a Sep-pak C18 column. Extracted LTB4 was derivatized into the pentafluorobenzyl ester of bis-trimethylsilyl ether (PFB-TMS-LTB4) and quantified on the basis of selected reaction monitoring (SRM) at m/z 299 of [M-PFB-2TMSOH]- by NICI/GC/MS/MS analysis, which was the product ion of [M-PFB]-. The detection limit for the quantification of LTB4 in human plasma was 10 pg ml-1, sufficiently sensitive to determine the concentrations of endogenous LTB4 in human plasma. The plasma level of LTB4 measured in healthy male volunteers was 33.85 +/- 33.91 pg ml-1 (mean +/- S.D. in six volunteers). The technique of MS/MS used in this method offers much greater sensitivity and selectivity than single-stage mass spectrometry. The developed method showed good reproducibility with a simple and rapid extraction procedure, and would be useful for examining the relationship between various disease states and the levels of LTB4 in biological fluids.

Topics: Fluorobenzenes; Gas Chromatography-Mass Spectrometry; Humans; Indicators and Reagents; Leukotriene B4; Male; Trimethylsilyl Compounds