Abstract
A highly sensitive and specific assay for the quantitation of prostaglandins (PGs) such as PGE 1, PGE 2, PGF 1α, PGF 2α, 6-keto-PGF 1α, and including thromboxane B 2, is described. The method involves the addition of PGF 1α and PGE 1 as the internal standards, extraction from whole blood and purification by silica gel column chromatography. Following conversion into the methoximes, purification by reversed-phase chromatography and esterification with panacyl bromide, samples are analysed by high-performance liquid chromatography with fluorimetric detection. The lower limit of detection of the eicosanoids 6-keto-PGF 1α, thromboxane B 2 and PGF 2α in blood is ca. 50 pg/ml and that of PGE 2 is 100 pg/ml. Assay linearity is demonstrated over a range from 60 pg to 60 ng of eicosanoid injected. The method allows simultaneous assessment of prostaglandins and thromboxane extracted from complex biological fluids at picogram levels.
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