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  • 1
    ISSN: 0887-6134
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: The aim of the present investigation was to ascertain whether mass spectrometric analysis of glucose allows determination in small samples (0.01 nmol) of the sites and the extent of labelling of glucose produced by isolated liver cells from various gluconeogenic labelled precursors. The electron impact spectrum of the methyloxime pentatrimethylsilyl derivative of natural glucose affords fragment ions retaining specific carbon atoms, i.e. 1-2 (m/z 160), 1-2-3 (m/z 262), 3-4-5-6 (m/z 319), 4-5-6 (m/z 217), 5-6 (m/z 205), 6 (m/z 103). The mass fragmentography analysis of the same derivative of commercially available labelled glucose molecules (1-13C, 6-13C, 2-2H, 3-2H, 6,6-2H2) permitted evaluation of the degree of specificity of these fragment ions, and development of a calculation method for isotope incorporation. Using this methodology we found that incubation of hepatocytes with (2-13C)glycerol, (1,3-13C)glycerol or NaH13CO3 plus pyruvate or lactate produced (2,5-13C)glucose, (1,3,4,6-13C) glucose or (3,4-13C)glucose, respectively. The extent of labelling was measurable on individual carbon of the glucose molecule except for carbon 1. The lowest enrichment detectable on carbon 1-3 or 3 was found to be 0.5%. In conclusion, gas chromatography mass spectrometry is a reliable method for positional isotopic anlysis of 13C-labelled glucose, and appears useful in the study of the gluconeogenic pathway.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1052-9306
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Chemistry and Pharmacology
    Notes: We present gas chromatographic/mass spectrometric and gas chromatographic/isotope ratio mass spectrometric assays of the 13C enrichment of plasma urea converted to its dimethylaminomethylene derivative. The limits of sensitivity of the two techniques are 0.2% and 0.02%, respectively. The techniques were tested in rats and humans infused with (13C)urea or (3-13C)lactate. (13C)Urea enrichment during the infusion of (3-13C)lactate in humans was not detectable by gas chromatography/mass spectrometry but was easily measured by gas chromatography/isotope ratio mass spectrometry. These assays should be useful for clinical investigations, in which the incorporation of a (13C)gluconeogenic substrate into glucose must be corrected for the incorporation of 13CO2 derived from the oxidation of the substrate. This correction involves measuring the low-level 13C enrichment of urea.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
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