Quantitative analysis of prostanoids in biological fluids by combined capillary column gas chromatography negative ion chemical ionization mass spectrometry

Abstract

An assay for the quantitative analysis of six biologically important prostanoids based on combined gas chromatography negative ion chemical ionization mass spectrometry has been developed. Prostanoids were extracted from biological fluids by liquid chromatography on Sep-Pak cartridges and converted to pentafluorobenzyl ester derivatives. Samples were injected on capillary column by the splitless technique and injections were made in a high boiling hydrocarbon solvent (n-dodecane) in order to minimize chromatographic run times. Quantification was carried out using selected ion monitoring of the appropriate [M-pentafluorobenzyl]- anion. The assay has been used for profiling cyclooxygenase metabolites of arachidonic acid in guinea pig lung perfusate after induction of anaphylaxis and platelet rich plasma after collagen-stimulated aggregation.

Release of arachidonic acid metabolites and histamine from sensitized guinea‐pig lung following antigen challenge

“…Prostanoids were extracted and derivatised by a modification of the method described by Waddell et al (1984). Aliquots of lung perfusate with added internal deuterated standards were made up to 10ml with equal volumes of distilled water and 3M acetate buffer, pH 5.2.…”

Effect of prostaglandin D2 on histamine‐induced weals in human skin

“…Order of injection was randomised, but corresponding injections of histamine alone or histamine with PGD2 were made at the same level either side of the midline. (Waddell et al, 1984). All dilutions of histamine and PGD2 were freshly made in sterile isotonic saline.…”

Glycosphingolipid Changes Induced by Advanced Glycation End-Products

“…GM3 was extracted from the silica by elution on small columns with C/M (1:1), C/M (1:2) and methanol. Free fatty acids of GM3 were obtained according to Aveldano and Horrocks (19) after incubation with 0.5 M HCl in acetonitrile/H 2 O (9:1, v/v) at 75°C for 2 h. FFA were derivatized as already described using pentafluorobenzene (PFB) and N,Obis(trimethylsilyl)trifluoroacetamide (BSTFA) (20). Free fatty acid derivatives were analyzed by GC-MS as already described (21).…”