Estimation of pentobarbital hydroxylase activity by gas-liquid chromatography

Jordan L. Holtzman, David S. Alberts

Research output: Contribution to journalArticle

6 Scopus citations

Abstract

A method is presented for the estimation by gas-liquid chromatography of the in vitro metabolism of pentobarbital to its alcoholic metabolite, hydroxypentobarbital (5-ethyl-5-(1′-methyl-3′-hydroxybutyl) barbituric acid). The parent compound and metabolite are extracted from the incubation medium with ethyl acetate, the ethyl acetate is removed, and the trimethylsilyl derivatives are formed. These are separated and the metabolite quantitated on a gas-liquid chromatograph with a hydrogen flame ionization detector. As little as 1% metabolism can be readily determined. The identification of the products was confirmed by thin-layer chromatography and combination gas-liquid chromatography-mass spectrography. The present method offers a compromise between the sensitivity of the method employing radiolabeled pentobarbital and the convenience of measuring the disappearance of substrate.

Original languageEnglish (US)
Pages (from-to)48-56
Number of pages9
JournalAnalytical Biochemistry
Volume43
Issue number1
DOIs
StatePublished - Sep 1971
Externally publishedYes

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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