A highly sensitive and selective method for the determination of Leukotriene B4 in human plasma by negative ion chemical ionization/gas chromatography/tandem mass spectrometry.
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.
Takamoto
M
M
Pharmacokinetic Research Laboratory, ONO Pharmaceutical Co., Ltd., Osaka, Japan.
Yano
T
T
Shintani
T
T
Hiraku
S
S
D005464:Fluorobenzenes; D008401:Gas Chromatography-Mass Spectrometry; D006801:Humans; D007202:Indicators and Reagents; D007975:Leukotriene B4 / Q000097:blood*; D008297:Male; D014297:Trimethylsilyl Compounds; D005466:Fluorocarbons
D005464:Fluorobenzenes; D007202:Indicators and Reagents; D007975:Leukotriene B4; D014297:Trimethylsilyl Compounds; C025063:pentafluorobenzyl bromide; D005466:Fluorocarbons