Spectrophotometric enzymatic assay for S-3-hydroxyisobutyrate

Paul M. Rougraff, Ralph Paxton, Gary W. Goodwin, Reid G. Gibson, Robert A. Harris

Research output: Contribution to journalArticle

9 Scopus citations

Abstract

An enzymatic spectrophotometric end-point assay has been developed for determination of S-3-hydroxyisobutyrate in biological fluids. The assay measures NADH production at 340 nm after initiation of the reaction with rabbit liver 3-hydroxyisobutyrate dehydrogenase (EC 1.1.1.31). The assay is not affected by R-3-hydroxyisobutyrate, lactate, malate, 3-hydroxybutyrate, 2-methyl-3-hydroxybutyrate, 3-hydroxyisovalerate, 3-hydroxy-n-valerate, 2-methyl-3-hydroxyvalerate, and 3-hydroxypropionate. The assay does measure 2-ethyl-3-hydroxypropionate, a minor metabolite produced by catabolism of alloisoleucine. Application of the method to measure S-3-hydroxyisobutyrate in plasma obtained from normal, 48-h starved, and mildly and severely diabetic rats gave levels of 28, 42, 112, and 155 μm, respectively.

Original languageEnglish (US)
Pages (from-to)317-320
Number of pages4
JournalAnalytical biochemistry
Volume184
Issue number2
DOIs
StatePublished - Feb 1 1990

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ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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