Hydroxycarboxylic and oxocarboxylic acids in urine: products from branched-chain amino acid degradation and from ketogenesis.

Hydroxy- and oxomonocarboxylic acids in urine of healthy individuals and of patients with diabetic ketoacidosis are analysed as methyl esters and methyl esters/O-methyloximes, respectively, by gas chromatography and gas chromatography-mass spectrometry. The derivatives are pre-fractionated by thin-layer chromatography. The acids originate mainly from ketogenesis and from the metabolism of valine, leucine and isoleucine. The amino acid metabolites fall into three groups: the 2-oxocarboxylic acids (2-oxoisovaleric acid, 2-oxoisocaproic acid and 2-oxo-3-methylvaleric acid); the 2-hydroxycarboxylic acids (2-hydroxyisovaleric acid, 2-hydroxyisocaproic acid and 2-hydroxy-3-methylvaleric acid); and the 3-hydroxycarboxylic acids (3-hydroxyisobutyric acid, 3-hydroxyisovaleric acid, 3-hydroxy-2-ethylpropionic acid, threo-3-hydroxy-2-methylbutyric acid and erythro-3-hydroxy-2-methylbutyric acid). The threo form of 3-hydroxy-2-methylbutyric acid is the major constituent within the diastereomeric pair. Of the three groups of amino acid metabolites, the 3-hydroxycarboxylic acids in particular are elevated during ketoacidosis. The characteristic general features of the mass spectrometric fragmentation of the derivatives of the identified components are systematically described. The discussion of the fragmentation includes constituents of low concentrations, such as 3-oxocaproic acid, 4-oxobutyric acid and 5-oxocaproic acid, which can be detected only when the pre-fractionation technique is applied.