Simultaneous determination of [2-15N]- and [5-15N]glutamine with gas chromatography-mass spectroscopy: applications to nitrogen metabolic studies |
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| Authors: | I Nissim M Yudkoff A Lapidot |
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| Affiliation: | 1. The Isotope Department, The Weizmann Institute of Science, 76100 Rehovot, Israel;2. Division of Biochemical Development and Molecular Disease, Children''s Hospital of Philadelphia, Philadelphia, Pennsylvania 19104 USA |
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| Abstract: | A gas chromatographic-mass spectrometric method for analysis of L-[2-15N]- and L-[5-15N]glutamine is described. The method is based on direct acylation of glutamine with trifluoroacetic anhydride and the formation of the N,N-bis-trifluoroacetyl-L-glutamine derivative. This simple and sensitive method is capable of detecting approximately 0.5 atom% excess 15N in as little as 10 microliter of plasma with a mean coefficient of variance of 11.6%. The method was applied to determine the appearance of 15N enrichment in plasma amino-N and amide-N of glutamine in a healthy adult volunteer during a constant infusion of 15NH4Cl. A plateau level of 3.7 and 2.6 atom% excess was observed in amide-N and amino-N, respectively, at 1 and 2 h after 15NH4Cl infusion was started. |
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| Keywords: | To whom all correspondence should be addressed. |
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