Simultaneous determinations of histamine and N tau-methylhistamine by high-performance liquid chromatography-chemiluminescence coupled with immobilized diamine oxidase.
Simultaneous determinations of histamine and N tau-methylhistamine by high-performance liquid chromatography-chemiluminescence coupled with immobilized diamine oxidase.
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固定化二胺氧化酶高效液相色谱-化学发光法同时测定组胺和N-tau-甲基组胺。
DOI：
10.1006/abio.1995.1374
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发表时间：
1995
影响因子：
2.9
通讯作者：
K. Maeyama
中科院分区：
文献类型：
--
作者：
M. K. Alam;M. Sasaki;T. Watanabe;K. Maeyama
建立了高效液相色谱-化学发光法同时测定组胺及其代谢产物N-牛甲基组胺的方法。该方法基于鲁米诺-铁氰化钠混合物在碱性介质中与过氧化氢反应生成的化学发光的测定，过氧化氢是组胺和N-牛甲基组胺在二胺氧化酶中形成的代谢产物之一。柱后衍生化高效液相色谱分离这两种胺类物质，两者的浓度与其化学发光强度呈线性关系。化学发光检测组胺和N-tau-甲基组胺的下限分别为5和10pmol。固定化柱在1个月以上的时间里表现出良好的运行稳定性，在此期间对200个样品进行了分析。用该系统测定Wistar大鼠大脑皮层、前胃、腺胃和肾脏的组胺含量分别为0.30、58、396和2.4nmol/g湿重。这些值与高效液相-荧光法测定的值非常相似。N-甲基组胺含量分别为0.36、0.40、0.72和3.8nmol/g湿重。这种方法将有助于研究组胺在脑和外周组织中的作用。
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.