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Title
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Validated comprehensive metabolomics and lipidomics analysis of colon tissue and cell lines
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Author
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Abstract
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| Current untargeted approaches for metabolic fingerprinting of colon tissue and cell lines lack validation of reproducibility and/or focus on a selection of metabolites as opposed to the entire metabolome. Yet, both are critical to ensure reliable results and pursue a fully holistic analysis. Therefore, we have optimized and validated a platform for analyzing the polar metabolome and lipidome of colon-derived cell and tissue samples based on a consecutive extraction of polar and apolar components. Peak areas of selected targeted analytes and the number of untargeted components were assessed. Analysis was performed using ultra-high performance liquid-chromatography (UHPLC) coupled to hybrid quadrupole-Orbitrap high-resolution mass spectrometry (HRMS). This resulted in an optimized extraction protocol using 50% methanol/ultrapure water to obtain the polar fraction followed by a dichloromethane-based lipid extraction. Using this comprehensive approach, we have detected more than 15,000 components with CV < 30% in internal quality control (IQC) samples and were able to discriminate the non-transformed (NT) and transformed (T) state in human colon tissue and cell lines based on validated OPLS-DA models (R2Y > 0.719 and Q2 > 0.674). To conclude, our validated polar metabolomics and lipidomics fingerprinting approach could be of great value to reveal gastrointestinal disease-associated biomarkers and mechanisms. |
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Language
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English
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Source (journal)
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Analytica chimica acta. - Amsterdam, 1947, currens
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Publication
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Amsterdam
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2019
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ISSN
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0003-2670
[print]
1873-4324
[online]
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DOI
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10.1016/J.ACA.2019.03.020
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Volume/pages
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1066
(2019)
, p. 79-92
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ISI
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000465212100009
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Pubmed ID
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31027537
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Full text (Publisher's DOI)
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Full text (open access)
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Full text (publisher's version - intranet only)
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