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Toward ‘Omic Scale Metabolite Profiling: A Dual Separation–Mass Spectrometry Approach for Coverage of Lipid and Central Carbon Metabolism

Ivanisevic, Julijana ; Zhu, Zheng-Jiang ; Plate, Lars ; Tautenhahn, Ralf ; Chen, Stephen ; O’Brien, Peter J ; Johnson, Caroline H ; Marletta, Michael A ; Patti, Gary J ; Siuzdak, Gary

Analytical chemistry (Washington), 2013-07, Vol.85 (14), p.6876-6884 [Periódico revisado por pares]

United States: American Chemical Society

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  • Título:
    Toward ‘Omic Scale Metabolite Profiling: A Dual Separation–Mass Spectrometry Approach for Coverage of Lipid and Central Carbon Metabolism
  • Autor: Ivanisevic, Julijana ; Zhu, Zheng-Jiang ; Plate, Lars ; Tautenhahn, Ralf ; Chen, Stephen ; O’Brien, Peter J ; Johnson, Caroline H ; Marletta, Michael A ; Patti, Gary J ; Siuzdak, Gary
  • Assuntos: Amino acids ; Bacteria ; Biological ; Burkitt Lymphoma - metabolism ; Carbon ; Carbon - analysis ; Carbon - metabolism ; Cells ; Chromatography ; Chromatography, Liquid - methods ; Computational chemistry ; Humans ; INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CHEMISTRY ; Ionization ; Lipid Metabolism - physiology ; Lipids ; Mass Spectrometry - methods ; Metabolism ; Metabolites ; Metabolomics - methods ; Molecules ; Phosphorylation ; Profiling ; Separation ; Solvents ; Tandem Mass Spectrometry - methods
  • É parte de: Analytical chemistry (Washington), 2013-07, Vol.85 (14), p.6876-6884
  • Notas: ObjectType-Article-1
    SourceType-Scholarly Journals-1
    ObjectType-Feature-2
    content type line 23
    National Institute on Aging (NIA)
    National Institutes of Health (NIH)
    AC02-05CH11231; FG02-07ER64325
    USDOE Office of Science (SC), Biological and Environmental Research (BER)
    California Institute of Regenerative Medicine
  • Descrição: Although the objective of any ‘omic science is broad measurement of its constituents, such coverage has been challenging in metabolomics because the metabolome is comprised of a chemically diverse set of small molecules with variable physical properties. While extensive studies have been performed to identify metabolite isolation and separation methods, these strategies introduce bias toward lipophilic or water-soluble metabolites depending on whether reversed-phase (RP) or hydrophilic interaction liquid chromatography (HILIC) is used, respectively. Here we extend our consideration of metabolome isolation and separation procedures to integrate RPLC/MS and HILIC/MS profiling. An aminopropyl-based HILIC/MS method was optimized on the basis of mobile-phase additives and pH, followed by evaluation of reproducibility. When applied to the untargeted study of perturbed bacterial metabolomes, the HILIC method enabled the accurate assessment of key, dysregulated metabolites in central carbon pathways (e.g., amino acids, organic acids, phosphorylated sugars, energy currency metabolites), which could not be retained by RPLC. To demonstrate the value of the integrative approach, bacterial cells, human plasma, and cancer cells were analyzed by combined RPLC/HILIC separation coupled to ESI positive/negative MS detection. The combined approach resulted in the observation of metabolites associated with lipid and central carbon metabolism from a single biological extract, using 80% organic solvent (ACN:MeOH:H2O 2:2:1). It enabled the detection of more than 30,000 features from each sample type, with the highest number of uniquely detected features by RPLC in ESI positive mode and by HILIC in ESI negative mode. Therefore, we conclude that when time and sample are limited, the maximum amount of biological information related to lipid and central carbon metabolism can be acquired by combining RPLC ESI positive and HILIC ESI negative mode analysis.
  • Editor: United States: American Chemical Society
  • Idioma: Inglês
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  • Realização: ABCD-USP USP   Logos de Redes Sociais: Freepik

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