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A rapid and sensitive method for measurement of leukotrienes based on HPLC
1Department of Physiological Chemistry, Karolinska Institute, Stockholm, Sweden.
Abstract:
A rapid and sensitive method for simultaneous detection of leukotriene (LT) B4, LTC4, LTD4, LTE4, 5S,12S-dihydroxy-eicosatetraenoic acid, 6-trans LTB4, 12-epi-6-trans LTB4 and the omega-metabolites of LTB4 has been developed. Leukocyte incubations were terminated by the addition of one volume of methanol, and the mixture was centrifuged. Analysis was then directly performed by reverse phase high performance liquid chromatography on a column packed with 3 microns Nucleosil C18 guarded by a precolumn. Quantitation of eluted compounds was carried out by UV-monitoring at 280 nm and integration of the elution profile. The sensitivity of the method was approximately 1 ng for both LTB4 and LTC4. Accuracy was 101.6% (SD +/- 9.8%) for LTB4 and 99.9% (SD +/- 8.7%) for LTC4 in the interval 5-160 ng.