Combined use of 1H-NMR and GC-MS for metabolite monitoring and in vivo 1H-NMR assignments

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149 Scopus citations

Abstract

Thirty-three metabolites were observed in perchloric acid extracts of four different tissues by in vitro 1H-NMR, GC-MS and alcohol dehydrogenase assay, and the information was used to interpret an in vivo two-dimensional nuclear Overhauser effect 1H-NMR spectrum. The metabolite profiles of the differt tissues indicate a number of potential tissue-specific markers: N-acetylaspartate and γ-aminobutyric acid for rat brain, glutamine/glutamic acid ratio for dog heart, arginine and sucrose for carrot, and t-aconitate, sucrose, aspragine/aspartic acid concentration ratios for corm roots. γ-Aminobutyric acid and malate can be regarded as metabolic indicators for stressed corn roots. Concentrations of threonine and valine in corn roots were constant under hypoxic and salt stress, and can serve as internal standards for both in vivo and in vitro NMR studies. The in vitro information was further used to identify 12 compounds from the in vivo 1H-NMR spectra (including the two-dimensional nuclear Overhauser effect spectrum) of a carrot cylinder by correlating the chemical shift and nuclear Overhauser effect information. Thus, our choice of methods with a capability for structural determination allows the characterization of complex tissue extracts with minimum sample preparation, and supports, as well as complements, in vivo 1H-NMR investigations of metabolism.

Original languageEnglish
Pages (from-to)154-167
Number of pages14
JournalBiochimica et Biophysica Acta - General Subjects
Volume882
Issue number2
DOIs
StatePublished - Jun 19 1986

Bibliographical note

Funding Information:
This work was supported in part by NIH Grant No. 007711 and NSF Grant DMB 8204877. A.N.L.

Funding Information:
is grateful to EMBO for a Long Term Fellowship. We thank Dr. D.J. Wilbur for his assistance in the early stages of this study, N. Wade-Jardetzky for sample handling of the mammalian tissues, and Dr. B.H. Walpoth and Dr. G. Elliott for providing the animal samples, and Professor E.E. Conn for useful comments.

Keywords

  • GC-MS
  • H-NMR
  • Metabolite monitoring
  • Perchlorate extract

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology

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