A sensitive method for simultaneous determination of histamine and noradrenaline with high-performance liquid chromatography/electrochemistry

Abstract

A simple and reliable high-performance liquid Chromatographic method is described for the simultaneous determination of histamine (His), which cannot be directly oxidized, and noradrenaline (NA), which can be directly oxidized within the useful working potential range. The isoindole products formed by precolumn derivatization of His and NA with o-phthalaldehyde (OPA) and 2-mercaptoethanol (2-ME) yielded a linear relationship of detection between the electrochemical signal and the compound content to a minimum detectable limit of 50 pg (signalto-noise ratio = 3:1) for both compounds at 0.5 nA of detector range. Without 2-ME, OPA derivatives of both His and NA were not detectable electrochemically at the oxidation potential range from 0 to +1 V. Although the peak potential was +0.85 V for both His and NA, we used +0.7 V for both compounds to keep background noise minimal. The capacity factors of some electrochemically interfering compounds were also determined. The significance of OPA/2-ME derivative of NA is discussed relative to the direct oxidation of catecholamines. An example of a practical application of the method to the determination of His and NA in rat cardiac tissue is presented.