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通过高效液相色谱法结合化学预衍生化荧光测定色氨酸和赖氨酸的常见代谢产物2-氧代己二酸。

Fluorometric determination of 2-oxoadipic acid, a common metabolite of tryptophan and lysine, by high-performance liquid chromatography with pre-chemical derivatization.

作者信息

Shibata Katsumi, Yasui Miki, Sano Mitsue, Fukuwatari Tsutomu

机构信息

Department of Food Science and Nutrition, School of Human Cultures, The University of Shiga Prefecture, 2500 Hassaka, Hikone, Shiga 522-8533, Japan.

出版信息

Biosci Biotechnol Biochem. 2011;75(1):185-7. doi: 10.1271/bbb.100723. Epub 2011 Jan 7.

Abstract

2-Oxoadipic acid, a key metabolite of tryptophan and lysine, reacted with 1,2-diamino-4,5-methylenebenzene in an acidic solution to produce a fluorescent derivative. The reaction product was separated using a Tosoh ODS-80Ts column with 20 mmol/L of KH₂PO₄-K₂HPO₄ buffer (pH 7.0) containing 26% methanol at a flow rate 0.8 mL/min. The excitation wavelength of detection was 367 nm, and the emission wavelength was 446 nm. The limit of quantification was 1 pmol per injection, sufficiently sensitive for the determination of 2-oxoadipic acid in human and experimental animal urine.

摘要

2-氧代己二酸是色氨酸和赖氨酸的关键代谢产物,它在酸性溶液中与1,2-二氨基-4,5-亚甲基苯反应生成一种荧光衍生物。使用Tosoh ODS-80Ts柱,以含26%甲醇的20 mmol/L KH₂PO₄-K₂HPO₄缓冲液(pH 7.0)为流动相,流速0.8 mL/min,对反应产物进行分离。检测的激发波长为367 nm,发射波长为446 nm。定量限为每次进样1 pmol,对测定人和实验动物尿液中的2-氧代己二酸具有足够的灵敏度。

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