A spectrophotometric determination of α-dicarbonyl compounds and its application to the enzymatic formation of α-ketobutyrate

Rongshi S. Li, George L. Kenyon

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

A simple, rapid, and sensitive spectrophotometric method has been developed for the determination of α-keto acids, α-diketones, and α-ketoaldehydes using o-phenylenediamine as a derivatizing reagent in acidic media to yield chromophoric quinoxaline derivatives. Application to the determination of α-ketobutyrate and the rate of its formation from isomerization of vinylglycolate catalyzed by mandelate racemase is described. Under optimal conditions 1 to 4 mM o-phenylenediamine in 80% acetic acid was incubated at 50°C for 10 min with the α-dicarbonyl compounds. Solutions of α-ketobutyrate as low as 5 μM in the buffer solution can be measured quantitatively. This method has also been used for determining the rate of formation of the phenylglyoxalate from (S)-mandelate catalyzed by (S)-mandelate dehydrogenase.

Original languageEnglish (US)
Pages (from-to)37-40
Number of pages4
JournalAnalytical Biochemistry
Volume230
Issue number1
DOIs
StatePublished - Sep 1995

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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