title: Application of reversed phase high performance liquid chromatography to the analysis of sulphidopeptide leukotrienes in pig bile.
abstract: Reversed phase HPLC methodology has been developed for separation of peptide leukotrienes and indomethacin in porcine bile. Reproducible recoveries were obtained using radioactive leukotrienes ([3H]LTC4, 57.1 +/- 2.5%; [3H]LTE4, 62.7 +/- 1.9%; [3H]LTD4, 54.3 +/- 2.7%). Radioimmunoassay of column eluant demonstrated that as little as 300 pg of exogenous leukotrienes could be measured in bile fluids, with similar recoveries. Analysis of bile sampled 60-90 min after initiation of experimental endotoxic shock in indomethacin treated pigs revealed a leukotriene concentration of 5.24 +/- 1.16 ng/mL(LTD4). This was significantly greater (p less than 0.05, n = 3) than that observed in samples collected prior to endotoxin (0.42 +/- 0.23 ng/mL), or from untreated animals (0.85 +/- 0.51 ng/mL). This method is thus applicable to investigation of the role of 5-lipoxygenase products in porcine models of human disease, including shock conditions such as endotoxaemia, during cyclooxygenase inhibition by indomethacin.
authors.affiliation: Department of Pharmacology, Merck Frosst Centre for Therapeutic Research, Pointe Claire-Dorval, Quebec, Canada.
authors.forename: P
authors.initials: P
authors.lastname: Tagari
authors.forename: D
authors.initials: D
authors.lastname: Ethier
authors.forename: A
authors.initials: A
authors.lastname: Ford-Hutchinson
authors.forename: M
authors.initials: M
authors.lastname: Cirino
mesh_terms: D000818:Animals; D001646:Bile / Q000032:analysis* / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005260:Female; D007213:Indomethacin / Q000494:pharmacology; D015289:Leukotrienes / Q000032:analysis* / Q000378:metabolism; D008297:Male; D013552:Swine
chemical_list: D015289:Leukotrienes; D007213:Indomethacin