Simultaneous determination of histamine and Nτ-methylhistamine with high-performance liquid chromatography using electrochemical detection

✍ Scribed by Kazunori Mine; Kenneth A. Jacobson; Kenneth L. Kirk; Yasuo Kitajima; Markku Linnoila

• Publisher: Elsevier Science
• Year: 1986
• Tongue: English
• Weight: 650 KB
• Volume: 152
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(86)90130-2

✦ Synopsis

We have developed a liquid chromatographic method which uses electrochemical detection for the simultaneous quantitation of histamine and N'-methylhistamine in rat brain. The amines are derivatized with the water-soluble Bolton-Hunter reagent (sulfo B-H). Perchloric acid extracts of rat brains are chromatographed on a strong cation-exchange resin. The eluate is evaporated and allowed to react with sulfa B-H at pH 9.8 at room temperature. The derivatization is complete after 30 s vortexing. The derivatives are purified using a cellulose-phosphate fibrous cation exchanger. They are quantified with an electrochemical detector at a potential of 0.56 V after preoxidizing the sample at 0.47 V. The derivatives of histamine, N'-methylhistamine, and N"methylhistamine are completely separated without interfering peaks. Since no N"-methylhistamine was detected in rat brain it was used as an internal standard. The detection limits are 0. I pmol of histamine and 0.2 pmol of N'-methylhistamine. The precision of this method is high, with within-run and between-run coefficients of variation of 2-7% and linearity of 0.999. Both histamine and N'-methylhistamine peak heights increased significantly and selectively after treatment with pargyline. Because of the high sensitivity, accuracy, and precision, the histamine and N'-methylhistamine contents of single nuclei of the rat hypothalamus can be routinely quantified.