Integrating comprehensive two-dimensional gas chromatography mass spectrometry and parallel two-dimensional liquid chromatography mass spectrometry for untargeted metabolomics - PubMed (original) (raw)

. 2019 Jul 21;144(14):4331-4341.

doi: 10.1039/c9an00560a. Epub 2019 Jun 13.

Affiliations

Integrating comprehensive two-dimensional gas chromatography mass spectrometry and parallel two-dimensional liquid chromatography mass spectrometry for untargeted metabolomics

Md Aminul Islam Prodhan et al. Analyst. 2019.

Abstract

The diverse characteristics and large number of entities make metabolite separation challenging in metabolomics. To date, there is not a singular instrument capable of analyzing all types of metabolites. In order to achieve a better separation for higher peak capacity and accurate metabolite identification and quantification, we integrated GC × GC-MS and parallel 2DLC-MS for analysis of polar metabolites. To test the performance of the developed system, 13 rats were fed different diets to form two animal groups. Polar metabolites extracted from rat livers were analyzed by GC × GC-MS, parallel 2DLC-MS (-) and parallel 2DLC-MS (+), respectively. By integrating all data together, 58 metabolites were detected with significant change in their abundance levels between groups (p≤ 0.05). Of the 58 metabolites, three metabolites were detected in two platforms and two in all three platforms. Manual examination showed that discrepancy of metabolite regulation measured by different platforms was mainly caused by the poor shape of chromatographic peaks resulting from low instrument response. Pathway analysis demonstrated that integrating the results from multiple platforms increased the confidence of metabolic pathway assignment.

PubMed Disclaimer

Figures

Figure 1.

Figure 1.

Overlap of metabolite identification. Metabolites were identified from GC×GC-MS by EI mass spectrum matching and retention index matching. (A) The metabolite identification in analysis of 2DLC-MS data was done by parent ion m/z matching. (B) The metabolite identification in analysis of 2DLC-MS/MS data by MS/MS spectrum matching with or without retention time match.

Figure 1.

Figure 1.

Overlap of metabolite identification. Metabolites were identified from GC×GC-MS by EI mass spectrum matching and retention index matching. (A) The metabolite identification in analysis of 2DLC-MS data was done by parent ion m/z matching. (B) The metabolite identification in analysis of 2DLC-MS/MS data by MS/MS spectrum matching with or without retention time match.

Figure 2.

Figure 2.

Overlap of metabolites detected with significant changes in their abundance levels between groups by three platforms.

Figure 3.

Figure 3.

Samples of instrument response of a metabolite affecting the quantification accuracy of that metabolite. (A) three dimensional chromatographic peak of

l-

ornithine detected by GC×GC-MS. (B) extracted ion chromatograms of

l-

ornithine in a randomly selected biological samples detected by 2DLC-MS (−). (C) Extracted ion chromatograms of

l-

ornithine in a randomly selected biological samples detected by 2DLC-MS (+). (D) three dimensional chromatographic peak of taurine detected by GC×GC-MS. (E) extracted ion chromatograms of taurine in a randomly selected biological samples detected by 2DLC-MS (−). (F) Extracted ion chromatograms of taurine in a randomly selected biological samples detected by 2DLC-MS (+).

Figure 4.

Figure 4.

Overlap of the pathways that were affected using different set of significant metabolites for metabolic pathway analysis.

References

    1. Sugimoto M, Wong DT, Hirayama A, Soga T and Tomita M, Metabolomics: Official journal of the Metabolomic Society, 2010, 6, 78–95. - PMC - PubMed
    1. Scalbert A, Brennan L, Fiehn O, Hankemeier T, Kristal BS, van Ommen B, Pujos-Guillot E, Verheij E, Wishart D and Wopereis S, Metabolomics: Official journal of the Metabolomic Society, 2009, 5, 435–458. - PMC - PubMed
    1. Kuehnbaum NL and Britz-McKibbin P, Chemical reviews, 2013, 113, 2437–2468. - PubMed
    1. Stoll DR and Carr PW, Anal. Chem, 2017, 89, 519–531. - PubMed
    1. Almstetter MF, Oefner PJ and Dettmer K, Analytical and bioanalytical chemistry, 2012, 402, 1993–2013. - PubMed

MeSH terms

Grants and funding

LinkOut - more resources