Fluorometric Determination of Indomethacin in Serum by High Performance Liquid Chromatography Using In-Line Oxidation with Hydrogen Peroxide

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Abstract
A fluorometric method has been developed for the determination of indomethacin In serum by high performance liquid chromatography. Indomethacin after deproteinization with acetonitriie was separated by reversed-phase chromatography with a neutral mobile phase (pH 7.0, 35% acetonitriie In phosphate buffer) containing hydrogen peroxide as a fluorogenic reagent, and was fluorometrically detected (Ex 358nm and Em 462nm) via in-line oxidation at high temperature (180 °C). The calibration curve was linear over the range of 2.5-15.0μg/ml by injecting a volume of 20μl of deproteinized serum. The detection limit (signal-to-noise ratio=3) for indomethacin in serum was 0.5μg/ml using a 20-μl aliquot of deproteinized serum.

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