Shibata Katsumi, Yasui Miki, Sano Mitsue, Fukuwatari Tsutomu
Department of Food Science and Nutrition, School of Human Cultures, The University of Shiga Prefecture, 2500 Hassaka, Hikone, Shiga 522-8533, Japan.
Biosci Biotechnol Biochem. 2011;75(1):185-7. doi: 10.1271/bbb.100723. Epub 2011 Jan 7.
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.
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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