High-performance liquid chromatographic method for profiling 2-oxo acids in urine and its application in evaluating vitamin status in rats |
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Authors: | Katsumi Shibata Chifumi Nakata Tsutomu Fukuwatari |
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Affiliation: | Department of Nutrition, School of Human Cultures, University of Shiga Prefecture, Hikone, Japan |
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Abstract: | 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 B1 and B6 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 30 mmol/L of KH2PO4 (pH 3.0):acetonitrile (7:3) at a flow rate of 1.0 mL/min. Vitamin B1 deficiency increased urinary levels of all 2-oxo acids, while vitamin B6 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. |
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Keywords: | urine 2-oxo acid 2-oxo acid-gram B-group vitamins nutritional status functional biomarker |
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