Fluorometric Determination of 2-Oxoadipic Acid, a Common Metabolite of Tryptophan and Lysine, by High-Performance Liquid Chromatography with Pre-Chemical Derivatization

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  • SHIBATA Katsumi
    Department of Food Science and Nutrition, School of Human Cultures, The University of Shiga Prefecture
  • YASUI Miki
    Department of Food Science and Nutrition, School of Human Cultures, The University of Shiga Prefecture
  • SANO Mitsue
    Department of Food Science and Nutrition, School of Human Cultures, The University of Shiga Prefecture
  • FUKUWATARI Tsutomu
    Department of Food Science and Nutrition, School of Human Cultures, The University of Shiga Prefecture

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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.

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