Enzymatic determination of the branched-chain alpha-keto acids.
A spectrophotometric endpoint assay for determination of branched-chain alpha-keto acids is described. The assay depends on measurement of the NADH produced after addition of branched-chain alpha-keto acid dehydrogenase. Interference by pyruvate and alpha-ketobutyrate was eliminated by pretreating the sample with pyruvate dehydrogenase. The method yielded a peripheral venous plasma value of 59 +/- 5 microM (mean +/- SE) for the branched-chain alpha-keto acids of five overnight fasted healthy humans.
D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide); D000328:Adult; D000818:Animals; D005260:Female; D006801:Humans; D007651:Keto Acids / Q000097:blood*; D007658:Ketone Oxidoreductases; D008297:Male; D009097:Multienzyme Complexes; D009243:NAD / Q000096:biosynthesis; D011768:Pyruvate Dehydrogenase Complex; D051381:Rats; D011919:Rats, Inbred Strains
D007651:Keto Acids; D009097:Multienzyme Complexes; D011768:Pyruvate Dehydrogenase Complex; D009243:NAD; D007658:Ketone Oxidoreductases; D042942:3-Methyl-2-Oxobutanoate Dehydrogenase (Lipoamide)