Mass spectrometric quantification of cysteine-containing leukotrienes in rat bile using 13C-labeled internal standards.
Leukotriene C4 and N-acetyl leukotriene E4 were determined in rat bile using a procedure based on high-performance liquid chromatographic fractionation, hydrogenation to 5-hydroxyeicosanoic acid, and gas chromatographic/mass spectrometric selected ion monitoring analysis of the pentafluorobenzyl ester, trimethylsilyl ether derivatives. 13C4-Labeled analogs of the leukotrienes were synthesized and used as internal standards. The concentrations of both leukotrienes in rat bile were markedly elevated after administration of endotoxin to anesthetized animals; N-acetyl leukotriene E4 was the more abundant product. The presence of leukotriene C4 in rat bile after endotoxin challenge was confirmed by fast atom bombardment/tandem mass spectrometry with precursor ion scanning. Quantitative determination of leukotriene C4 using the tandem mass spectrometric procedure was consistent with the gas chromatographic/mass spectrometric data but the latter procedure gave a substantially lower detection limit.
Raftery
M J
MJ
Center for Experimental Therapeutics, Baylor College of Medicine, Houston, Texas 77030.
Justesen
U
U
Jaeschke
H
H
Gaskell
S J
SJ
D000818:Animals; D001646:Bile / Q000737:chemistry*; D002247:Carbon Isotopes; D003545:Cysteine / Q000032:analysis*; D004731:Endotoxins / Q000494:pharmacology; D008401:Gas Chromatography-Mass Spectrometry; D015289:Leukotrienes / Q000032:analysis*; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008297:Male; D013058:Mass Spectrometry; D051381:Rats; D011916:Rats, Inbred F344; D016339:Spectrometry, Mass, Fast Atom Bombardment
D002247:Carbon Isotopes; D004731:Endotoxins; D015289:Leukotrienes; D003545:Cysteine