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Abstract
Metabolomic and lipidomic studies measure and discover metabolic and lipid profiles in biological samples, enabling a better understanding of the metabolism of specific biological phenotypes. Accurate biological interpretations require high analytical reproducibility and sensitivity, and standardized and transparent data processing. Here we describe a complete workflow for nanoelectrospray ionization (nESI) direct-infusion mass spectrometry (DIMS) metabolomics and lipidomics. After metabolite and lipid extraction from tissues and biofluids, samples are directly infused into a high-resolution mass spectrometer (e.g., Orbitrap) using a chip-based nESI sample delivery system. nESI functions to minimize ionization suppression or enhancement effects as compared with standard electrospray ionization (ESI). Our analytical technique-named spectral stitching-measures data as several overlapping mass-to-charge (m/z) windows that are subsequently 'stitched' together, creating a complete mass spectrum. This considerably increases the dynamic range and detection sensitivity-about a fivefold increase in peak detection-as compared with the collection of DIMS data as a single wide mass-to-charge (m/z ratio) window. Data processing, statistical analysis and metabolite annotation are executed as a workflow within the user-friendly, transparent and freely available Galaxy platform (galaxyproject.org). Generated data have high mass accuracy that enables molecular formulae peak annotations. The workflow is compatible with any sample-extraction method; in this protocol, the examples are extracted using a biphasic method, with methanol, chloroform and water as the solvents. The complete workflow is reproducible, rapid and automated, which enables cost-effective analysis of >10,000 samples per year, making it ideal for high-throughput metabolomics and lipidomics screening-e.g., for clinical phenotyping, drug screening and toxicity testing.
Original language | English |
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Pages (from-to) | 310-328 |
Journal | Nature protocols |
Volume | 12 |
Issue number | 2 |
DOIs | |
Publication status | Published - 12 Jan 2017 |
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Dive into the research topics of 'A complete workflow for high-resolution spectral-stitching nanoelectrospray direct-infusion mass-spectrometry-based metabolomics and lipidomics'. Together they form a unique fingerprint.Projects
- 2 Finished
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OMICS Discipline Hopping - Implementing Metabolomics Analyses into Galaxy Workflows: Towards Genome-Metabolome Large-Scale Data Fusion
Viant, M. (Principal Investigator) & Colbourne, J. (Co-Investigator)
Natural Environment Research Council
17/04/13 → 31/10/13
Project: Research Councils
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From Airborne Exposures to Biological Effects(FABLE): The Impact of Nanoparticles on Health
Viant, M. (Principal Investigator), Ayres, J. (Co-Investigator), Chang, I. (Co-Investigator), Delgado Saborit, J. M. (Co-Investigator), Lead, J. (Co-Investigator), Preece, J. (Co-Investigator), Rappoport, J. (Co-Investigator), Hodges, N. (Co-Investigator) & Knight, T. (Co-Investigator)
Natural Environment Research Council
1/05/11 → 31/10/15
Project: Research Councils