Histamine in tissue: determination by high-performance liquid chromatography after condensation with o-phthaldialdehyde

Abstract

Histamine was determined by reversed-phase high-performance liquid chromatography in perchloric acid extracts after condensation with o-phthaldialdehyde. Fluorescence was monitored at 360 nm excitation and 450 nm emission wavelengths after elution with mixtures of 0.1 N acetic acid containing 0.1% pentanesulfonic acid and acetonitrile. The detection limit was 0.9 pmol of histamine. The histamine content was determined in rat whole brain, medulla oblongata, dorsal and ventral spinal cord, dorsal and ventral skin of the hind paw, stomach, ileum, rectum, lung and a hind-quarter perfusate, and compared to published data. The advantages of the described method over other methods are (A) rapid analysis in an automated system, (B) no selective extraction procedure is necessary, and (C) interfering substances are easily separated from the histamine fluorophore.