Estimation of pentobarbital hydroxylase activity by gas-liquid chromatography

Jordan L. Holtzman, David S Alberts

Research output: Contribution to journalArticle

6 Citations (Scopus)

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 - 1971
Externally publishedYes

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Liquid chromatography
Metabolites
Gas chromatography
Gas Chromatography
Pentobarbital
Metabolism
Flame Ionization
Thin layer chromatography
Thin Layer Chromatography
Ionization
Hydrogen
Gases
Derivatives
Detectors
Liquids
Substrates
pentobarbital hydroxylase
ethyl acetate

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Estimation of pentobarbital hydroxylase activity by gas-liquid chromatography. / Holtzman, Jordan L.; Alberts, David S.

In: Analytical Biochemistry, Vol. 43, No. 1, 1971, p. 48-56.

Research output: Contribution to journalArticle

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