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Use of chemical ionization for GC–MS metabolite profiling

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Abstract

Metabolite profiling is commonly performed by GC–MS of methoximated trimethylsilyl derivatives. The popularity of this technique owes much to the robust, library searchable spectra produced by electron ionization (EI). However, due to extensive fragmentation, EI spectra of trimethylsilyl derivatives are commonly dominated by trimethylsilyl fragments (e.g. m/z 73 and 147) and higher m/z fragment ions with structural information are at low abundance. Consequently different metabolites can have similar EI spectra, and this presents problems for identification of “unknowns” and the detection and deconvolution of overlapping peaks. The aim of this work is to explore use of positive chemical ionization (CI) as an adjunct to EI for GC–MS metabolite profiling. Two reagent gases differing in proton affinity (CH4 and NH3) were used to analyse 111 metabolite standards and extracts from plant samples. NH3-CI mass spectra were simple and generally dominated by [MH]+ and/or the adduct [M+NH4]+. For the 111 metabolite standards, m/z 73 and 147 were less than 3% of basepeak in NH3-CI and less than 30% of basepeak in CH4-CI. With CH4-CI, [MH]+ was generally present but at lower relative abundance than for NH3-CI. CH4-CI spectra were commonly dominated by losses of CH4 [M+1-16]+, 1–3 TMSOH [M+1-nx90]+, and combinations of CH4 and TMSOH losses [M+1-nx90-16]+. CH4-CI and NH3-CI mass spectra are presented for 111 common metabolites, and CI is used with real samples to help identify overlapping peaks and aid identification via determination of the pseudomolecular ion with NH3-CI and structural information with CH4-CI.

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Acknowledgments

This work was supported by a Discovery Grant and QEII Fellowship from the Australian Research Council (DP0662752). Georgia Warren assisted with collection of plant samples.

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  1. School of Biological Sciences, The University of Sydney, Heydon-Laurence Building A08, Sydney, NSW, 2006, Australia

    Charles R. Warren

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  1. Charles R. Warren
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Correspondence to Charles R. Warren.

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Supplementary Table 1

Relative abundance of major fragment ions in mass spectra of methoximated trimethylsilyl derivatives of metabolites analysed by CH4-CI. Pubchem and CHEBI identifiers are given for all metabolites. Abbreviations are: RI = retention index; BP = basepeak; MH+ ** = MH+ or largest fragment ion with abundance >1%. To simplify presentation only abundant or structurally informative fragment ions are shown (XLS 93 kb)

Supplementary File 1

Library of CH4-CI mass spectra of methoximated trimethylsilyl derivatives of 111 metabolites. Mass spectra were obtained as described in the text. The library is in JCAMP-DX format to enable upload to other software tools (JDX 123 kb)

Supplementary File 2

Library of CH4-CI mass spectra of 21 n-alkanes. Mass spectra were obtained as described in the text. The library is in JCAMP-DX format to enable upload to other software tools (JDX 16 kb)

Supplementary File 3

Library of NH3-CI mass spectra of methoximated trimethylsilyl derivatives of 111 metabolites. Mass spectra were obtained as described in the text. The library is in JCAMP-DX format to enable upload to other software tools (JDX 104 kb)

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Warren, C.R. Use of chemical ionization for GC–MS metabolite profiling. Metabolomics 9 (Suppl 1), 110–120 (2013). https://doi.org/10.1007/s11306-011-0346-8

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