Alam M K, Sasaki M, Watanabe T, Maeyama K
Department of Pharmacology I, Tohoku University School of Medicine, Sendai, Japan.
Anal Biochem. 1995 Jul 20;229(1):26-34. doi: 10.1006/abio.1995.1374.
A method for the simultaneous determinations of histamine and its metabolite N tau-methylhistamine by HPLC-chemiluminescence coupled with immobilized diamine oxidase was developed. The method was based on the determination of chemiluminescence formed by the reaction of a luminol-ferricyanide mixture in alkaline medium with hydrogen peroxide which is one of the metabolic products of histamine and N tau-methylhistamine formed by diamine oxidase. HPLC with postcolumn derivatization resulted in good separation of the two amines and gave linear relationships between the concentrations of both and their chemiluminescence intensities. The lower limits of chemiluminescent detection of histamine and N tau-methylhistamine were 5 and 10 pmol, respectively. The immobilized column showed good operational stability for more than 1 month, during which period 200 samples were analyzed. With this system, the histamine contents of the cerebral cortex, forestomach, glandular stomach, and kidney of Wistar rats were found to be 0.30, 58, 396, and 2.4 nmol/g wet wt, respectively. These values are very similar to those determined by HPLC-fluorometry. The N tau-methylhistamine contents of these tissues were 0.36, 0.40, 0.72, and 3.8 nmol/g wet wt, respectively. This method will be useful for studying the roles of histamine in both brain and peripheral tissues.
建立了一种通过高效液相色谱-化学发光联用固定化二胺氧化酶同时测定组胺及其代谢产物N-τ-甲基组胺的方法。该方法基于在碱性介质中，鲁米诺-铁氰化物混合物与过氧化氢反应产生的化学发光的测定，而过氧化氢是组胺和二胺氧化酶形成的N-τ-甲基组胺的代谢产物之一。柱后衍生的高效液相色谱法实现了两种胺的良好分离，并给出了两者浓度与其化学发光强度之间的线性关系。组胺和N-τ-甲基组胺的化学发光检测下限分别为5和10 pmol。固定化柱在1个多月的时间内表现出良好的操作稳定性，在此期间分析了200个样品。使用该系统，发现Wistar大鼠大脑皮层、前胃、腺胃和肾脏中的组胺含量分别为0.30、58、396和2.4 nmol/g湿重。这些值与通过高效液相色谱-荧光法测定的值非常相似。这些组织中N-τ-甲基组胺的含量分别为0.36、0.40、0.72和3.8 nmol/g湿重。该方法将有助于研究组胺在脑和外周组织中的作用。
