Determination of glyoxylic acid in urine by liquid chromatography with fluorescence detection, using a novel derivatization procedure based on the Petasis reaction

Chihara, Kohki; Kishikawa, Naoya; Ohyama, Kaname; Nakashima, Kenichiro; Kuroda, Naotaka
November 2012
Analytical & Bioanalytical Chemistry;Nov2012, Vol. 403 Issue 9, p2765
Academic Journal
The Petasis reaction is the multi-component reaction of a carbonyl compound, amine, and arylboronic acid to form an α-amino acid or a β-aminoalcohol. In this work, as the first analytical application of the Petasis reaction, a high-performance liquid chromatographic (HPLC) method with fluorescence detection was developed for determination of glyoxylic acid. The glyoxylic acid was derivatized with 1-pyreneboronic acid, as fluorescent arylboronic acid, in the presence of N-methylbutylamine, as amine, to give a fluorescent α-amino acid. HPLC separation of the fluorescent derivative was performed within 30 min on an octyl column eluted with a gradient prepared from acetonitrile and 50 mmol L acetate buffer (pH 4.0). The detection limit ( S/ N=3) for glyoxylic acid was 5.0 nmol L (20 fmol/injection). The method can be used to determine the concentration of glyoxylic acid in human urine without interference from biological components.


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