Assessment of
eicosanoid levels in
biological systems is important for understanding their role in cell function and pathophysiological events. Current methods of
eicosanoid quantitation are limited by sensitivity, scope, or throughput. The development of a new method for
eicosanoid assessment in
biological samples by electrospray and tandem mass spectrometry (MS/MS) in the multiple reaction monitoring mode is described here. In this study, 14 biologically significant
eicosanoids were quantitated in a single sample. Complete sample analysis required two repeated
injections of 5 microliters with an analysis time of 1.5 min/injection. Limits of detection ranged from 0.5 pg for
thromboxane B2 (TxB2) to 10 pg for 6-keto
prostaglandin F1 alpha (
6-keto PGF1 alpha). The reliability, reproducibility, sensitivity, and cross-detection of the method is also described. The MS/MS method was used to explore
eicosanoid production in two
inflammation models:
lipopolysaccharide (LPS)-stimulated human whole blood and
carrageenan-challenged rat air pouch. The most abundant metabolites in LPS-stimulated whole blood were
prostaglandin E2 (
PGE2), TxB2, and 6-keto
PGF1 alpha;
prostaglandins E1, D2, and F2 alpha and
leukotrienes B4 and C4 were detected in lower amounts.
Eicosanoid levels determined by MS/MS were similar to those obtained by immunoassay and GC-MS. The most abundant metabolites detected in
carrageenan-challenged rat air pouch were
PGE2, 6-keto
PGF1 alpha, and TxB2. The method described in this work is accurate and rapid and should greatly aid in evaluating the role of multiple
eicosanoids in future
biological studies.