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人血浆中芳香族L-氨基酸脱羧酶的存在及其用荧光检测高效液相色谱法进行测定

Occurrence of aromatic L-amino acid decarboxylase in human plasma and its assay by high-performance liquid chromatography with fluorescence detection.

作者信息

Lee M, Nohta H, Ohkura Y

出版信息

J Chromatogr. 1986 Jun 13;378(2):329-36. doi: 10.1016/s0378-4347(00)80729-x.

Abstract

A high-performance liquid chromatographic method using fluorescence detection for assessing the activity of aromatic L-amino acid decarboxylase in human plasma is described. Dopamine, formed enzymatically from L-DOPA, and isoproterenol (internal standard) are chromatographed on a small ion-exchange cartridge (Toyopak SP) and derivatized with 1,2-diphenylethylenediamine. The derivatives are separated by reversed-phase chromatography on an Ultrasphere ODS column. The detection limit for dopamine formed enzymatically is 0.6 pmol per 500 microliter of enzyme reaction mixture. Aromatic L-amino acid decarboxylase in human plasma is very similar to that in rat kidney, with respect to optimum conditions for the enzyme reaction and gel chromatographic behaviour.

摘要

描述了一种使用荧光检测的高效液相色谱法,用于评估人血浆中芳香族L-氨基酸脱羧酶的活性。由L-多巴酶促形成的多巴胺和异丙肾上腺素(内标)在小型离子交换柱(Toyopak SP)上进行色谱分离,并用1,2-二苯基乙二胺衍生化。衍生物在Ultrasphere ODS柱上通过反相色谱法分离。酶促形成的多巴胺的检测限为每500微升酶反应混合物0.6皮摩尔。就酶反应的最佳条件和凝胶色谱行为而言,人血浆中的芳香族L-氨基酸脱羧酶与大鼠肾脏中的非常相似。

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