An assay for plasma prostaglandins from bronchial asthmatic patients using 9-anthryldiazomethane-HPLC method.

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Prostaglandins (PGs) are generated by the lung with physiologic stimuli. There are many evidences that PGs are involved in immediate hypersensitivity reactions and their pathophysiologic roles in bronchial asthma have been suggested. Even then, the significance of their roles has not yet been established at the in vivo level in clinical studies.We describe here an efficient method for simultaneous and quantitative determinations of plasma PGs from bronchial asthmatic patients using reversed-phase HPLC.An overnight reaction of plasma PGs with 9-anthryldiazomethane (ADAM) was performed at room temperature. The mixture was loaded on GPC column to avoid interferences for the assay and PG-ADAM fraction was collected. The fraction was loaded on ODS column and was eluted with water-methanol gradient at 40°C. Fluorescence of PG-ADAM was measured in fluorometer with excitation at 350nm and emission at 412nm. Detection and simultaneous quantitation of 20pg PGE1, PGE2 and 100pg 6-keto PGF1α, thromboxane B2 was possible.