High-performance liquid chromatographic method for profiling 2-oxo acids in urine and its application in evaluating vitamin status in rats

作者：Shibata Katsumi^*; Nakata Chifumi; Fukuwatari Tsutomu

来源：Bioscience Biotechnology and Biochemistry, 2016, 80(2): 304-312.

DOI：10.1080/09168451.2015.1083395

• urine 2-oxo acid
• 2-oxo acid-gram
• B-group vitamins
• nutritional status
• functional biomarker

B-group vitamins are involved in the catabolism of 2-oxo acids. To identify the functional biomarkers of B-group vitamins, we developed a high-performance liquid chromatographic method for profiling 2-oxo acids in urine and applied this method to urine samples from rats deficient in vitamins B-1 and B-6 and pantothenic acid. 2-Oxo acids were reacted with 1,2-diamino-4,5-methylenebenzene to produce fluorescent derivatives, which were then separated using a TSKgel ODS-80Ts column with 30mmol/L of KH2PO4 (pH 3.0):acetonitrile (7:3) at a flow rate of 1.0mL/min. Vitamin B-1 deficiency increased urinary levels of all 2-oxo acids, while vitamin B-6 deficiency only increased levels of sum of 2-oxaloacetic acid and pyruvic acid, and pantothenic acid deficiency only increased levels of 2-oxoisovaleric acid. Profiles of 2-oxo acids in urine samples might be a non-invasive way of clarifying the functional biomarker of B-group vitamins.

• 出版日期2016-2-1