Add time:08/26/2019         Source:sciencedirect.com

A method for the fluorometric determination of α-ketosuccinamic acid, the α-keto acid analog of asparagine, is described. The procedure involves the hydrolysis of α-ketosuccinamate to oxaloacetate by ω-amidase followed by NADH-dependent reduction of oxaloacetate to malate by malate dehydrogenase. A correction for endogenous oxaloacetate is made by using control samples lacking ω-amidase. Of the rat tissues investigated, liver contained the highest concentration, followed by kidney (53 ± 6 (n = 11) and 18 ± 3 (n = 3) μmol/kg wet wt, respectively). α-Ketosuccinamate was not detected in brain (<8 μmol/kg wet wt). Some chemical properties of α-ketosuccinamate were investigated. Concentrated solutions of sodium α-ketosuccinamate frozen for extended periods and the solid sodium salt of α-ketosuccinamate dimer heated to 130°C are converted to at least 10 products by processes involving dimerization, dehydration, and decarboxylation. Isobutane chemical ionization mass spectral analysis (170–230°C) of the free acid monomer yielded similar products. Many of the breakdown products were identified as di- and monoheterocyclic compounds, some of which are known to be of biological importance.

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