Determination of capsaicin and zucapsaicin in human serum by high-performance liquid chromatography with fluorescence detection.

A reversed-phase high-performance liquid chromatographic (HPLC) assay was developed to analyze capsaicin and zucapsaicin (civamide) in human serum at concentrations from 1 to 100 ng/ml. Human serum specimens were extracted twice with hexane-methyl tert.-butyl ether (1:1). The chromatographic separation was carried out on a C18 column at 40 degrees C using a mobile phase consisting of 40% acetonitrile in water with 5% tetrahydrofuran and 1% acetic acid. The concentration of the eluting compounds was monitored by a fluorescence detector with excitation at 270 nm and an emission cutoff of 300 nm. No interferences were observed from the extract of blank serum. The standard curves were linear in the detection range. The relative standard deviation of the assay was better than 8.4%. The limit of detection was 0.5 ng/ml.