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正常和恶性高热猪血液中生物胺的柱前衍生化及高效液相色谱分析

Pre-column derivatization and high-performance liquid chromatography of biogenic amines in blood of normal and malignant hyperthermic pigs.

作者信息

Davis T P, Gehrke C W, Williams C H, Gehrke C W, Gerhardt K O

出版信息

J Chromatogr. 1982 Mar 12;228:113-22. doi: 10.1016/s0378-4347(00)80424-7.

Abstract

A sensitive, selective, pre-column derivatization method was used with high-performance liquid chromatography to measure norepinephrine, dopamine and serotonin in plasma from normal and malignant hyperthermic (MH) pigs. Samples were carefully collected from control and stressed animals under halothane anesthesia. Using a simple extraction method involving pre-column derivatization with o-phthaladehyde and ethyl acetate partitioning, the samples were chromatographed in less than 50 min. Norepinephrine was found to be elevated in MH pigs as the syndrome progressed, reaching levels eight-fold greater than control pigs under anesthesia. These experiments provide some evidence for our hypothesis that a failure to metabolize excess norepinephrine may be one of the key metabolic defects in causing the pathophysiology of the malignant hyperthermia-stress syndrome. The application of our chromatographic method in animal and human tests may provide a pattern of biogenic amine types and levels that could be diagnostic in identifying susceptible humans and carrier animals.

摘要

采用一种灵敏、选择性的柱前衍生化方法结合高效液相色谱法,测定正常猪和恶性高热(MH)猪血浆中的去甲肾上腺素、多巴胺和5-羟色胺。在氟烷麻醉下,从对照动物和应激动物中仔细采集样本。采用一种简单的提取方法,该方法涉及邻苯二甲醛柱前衍生化和乙酸乙酯分配,样本在不到50分钟内完成色谱分析。发现随着综合征的进展,MH猪体内的去甲肾上腺素升高,在麻醉状态下其水平比对照猪高八倍。这些实验为我们的假设提供了一些证据,即不能代谢过量的去甲肾上腺素可能是导致恶性高热-应激综合征病理生理学的关键代谢缺陷之一。我们的色谱方法在动物和人体测试中的应用可能会提供一种生物胺类型和水平的模式,这对于识别易感人群和携带动物具有诊断价值。

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