The determination of low d5-phenylalanine enrichment (0.002-0.09 atom percent excess), after conversion to phenylethylamine, in relation to protein turnover studies by gas chromatography/electron ionization mass spectrometry

Alexander Graham Calder, S E Anderson, I Grant, M A McNurlan, P J Garlick

    Research output: Contribution to journalArticle

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    Abstract

    A gas chromatography/mass spectrometry (GC/MS) method for measuring very low levels of enrichment of d5-phenylalanine (0.002-0.09 atom percent excess) is described. This method makes it possible to determine the enrichment of amino acid incorporated into tissue protein during studies of protein synthesis in man. Phenylalanine is enzymatically converted to phenylethylamine and the d5-enrichment is measured in the heptafluorobutyryl derivative by selective-ion recording under electron ionization conditions. The coefficients of variation for muscle-protein hydrolysate samples enriched with d5-phenylalanine at the 0.005 and 0.05 atom percent excess levels were 6.0 and 1.2%, respectively. This precision at low enrichment and the small amount of protein needed (about 1 mg) provide real advantages for clinical studies of tissue protein synthesis. Moreover, in contrast to the conventional approach which uses GC/MS for plasma amino acids (typically 2-20 atom percent excess) but gas isotope-ratio mass spectrometry for protein-bound amino acids, the enrichment of both plasma-free and protein-bound d5-phenylalanine can be measured with a single instrument.
    Original languageEnglish
    Pages (from-to)421-424
    Number of pages4
    JournalRapid Communications in Mass Spectrometry
    Volume6
    Issue number7
    DOIs
    Publication statusPublished - 1 Jul 1992

    Fingerprint

    Phenethylamines
    Phenylalanine
    Gas chromatography
    Ionization
    Mass spectrometry
    Atoms
    Electrons
    Proteins
    Amino Acids
    Tissue
    Protein Hydrolysates
    Plasmas
    Muscle Proteins
    Isotopes
    Gases
    Ions
    Derivatives

    Keywords

    • Gas Chromatography-Mass Spectrometry
    • Humans
    • Muscle Proteins
    • Phenethylamines
    • Phenylalanine

    Cite this

    The determination of low d5-phenylalanine enrichment (0.002-0.09 atom percent excess), after conversion to phenylethylamine, in relation to protein turnover studies by gas chromatography/electron ionization mass spectrometry. / Calder, Alexander Graham; Anderson, S E; Grant, I; McNurlan, M A; Garlick, P J.

    In: Rapid Communications in Mass Spectrometry, Vol. 6, No. 7, 01.07.1992, p. 421-424.

    Research output: Contribution to journalArticle

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    abstract = "A gas chromatography/mass spectrometry (GC/MS) method for measuring very low levels of enrichment of d5-phenylalanine (0.002-0.09 atom percent excess) is described. This method makes it possible to determine the enrichment of amino acid incorporated into tissue protein during studies of protein synthesis in man. Phenylalanine is enzymatically converted to phenylethylamine and the d5-enrichment is measured in the heptafluorobutyryl derivative by selective-ion recording under electron ionization conditions. The coefficients of variation for muscle-protein hydrolysate samples enriched with d5-phenylalanine at the 0.005 and 0.05 atom percent excess levels were 6.0 and 1.2{\%}, respectively. This precision at low enrichment and the small amount of protein needed (about 1 mg) provide real advantages for clinical studies of tissue protein synthesis. Moreover, in contrast to the conventional approach which uses GC/MS for plasma amino acids (typically 2-20 atom percent excess) but gas isotope-ratio mass spectrometry for protein-bound amino acids, the enrichment of both plasma-free and protein-bound d5-phenylalanine can be measured with a single instrument.",
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