title: Alkylation or Silylation for Analysis of Amino and Non-Amino Organic Acids by GC-MS?
abstract: Gas chromatography-mass spectrometry (GC-MS) is a widely used analytical technique in metabolomics. GC provides the highest resolution of any standard chromatographic separation method, and with modern instrumentation, retention times are very consistent between analyses. Electron impact ionization and fragmentation is generally reproducible between instruments and extensive libraries of spectra are available that enhance the identification of analytes. The major limitation is the restriction to volatile analytes, and hence the requirement to convert many metabolites to volatile derivatives through chemical derivatization. Here we compared the analytical performance of two derivatization techniques, silylation (TMS) and alkylation (MCF), used for the analysis of amino and non-amino organic acids as well as nucleotides in microbial-derived samples. The widely used TMS derivatization method showed poorer reproducibility and instability during chromatographic runs while the MCF derivatives presented better analytical performance. Therefore, alkylation (MCF) derivatization seems to be preferable for the analysis of polyfunctional amines, nucleotides and organic acids in microbial metabolomics studies.
authors: Centre for Microbial Innovation, School of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland, New Zealand. s.villas-boas@auckland.ac.nz.
authors: Silas G
authors: SG
authors: Villas-Bôas
authors: Centre for Microbial Innovation, School of Biological Sciences, The University of Auckland, Private Bag 92019, Auckland, New Zealand.
authors: Kathleen F
authors: KF
authors: Smart
authors: AgResearch Limited, Grasslands Research Centre, Tennent Drive, Palmerston North 4442, New Zealand.
authors: Subathira
authors: S
authors: Sivakumaran
authors: AgResearch Limited, Grasslands Research Centre, Tennent Drive, Palmerston North 4442, New Zealand.
authors: Geoffrey A
authors: GA
authors: Lane