title: Quantitative analysis of amino and organic acids by methyl chloroformate derivatization and GC-MS/MS analysis.
abstract: Alkyl chloroformates are known for their ability to produce mixed anhydrides, and they have found use as versatile derivatization reagents for gas chromatographic (GC) separation of amino- and organic acids. Triple-quadrupole mass spectrometers are excellent detectors for high sensitive and selective analysis. Here, we describe a methyl chloroformate (MCF) GC-MS/MS method for the quantitative analysis of metabolites containing amino- and/or carboxylic groups. The method covers over 60 metabolites with quantitation limits down to the low picomole range injected on column, and any metabolite with amino- and/or carboxylic acid functional groups that yield a stable and volatile MCF derivative can be included in the method. Absolute quantitation can be achieved by including a stable isotope-coded derivatization agent (d3-MCF) and deuterated alcohol solvent (e.g., d4-methanol). As the carboxylic and amino groups are differently labeled, the former from the solvent methanol while the latter from MCF, this approach can also be used to identify a number of amino and carboxylic groups in unknown analytes in an extract.
authors.affiliation: NTNU Department of Biotechnology, Norwegian University of Science and Technology, Sem Selands vei 6/8, 7491, Trondheim, Norway.
authors.forename: Hans Fredrik Nyvold
authors.initials: HF
authors.lastname: Kvitvang
authors.forename: Kåre A
authors.initials: KA
authors.lastname: Kristiansen
authors.forename: Stina K
authors.initials: SK
authors.lastname: Lien
authors.forename: Per
authors.initials: P
authors.lastname: Bruheim
mesh_terms: D000596:Amino Acids / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D002264:Carboxylic Acids / Q000032:analysis* / Q000378:metabolism; D005561:Formates / Q000737:chemistry*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D006801:Humans; D055442:Metabolome; D055432:Metabolomics / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D000596:Amino Acids; D002264:Carboxylic Acids; D005561:Formates; C014667:methyl chloroformate