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UHPLC-QTOF-MS/MS-SPE-NMR: A Solution to the Metabolomics Grand Challenge of Higher-Throughput, Confident Metabolite Identifications

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NMR-Based Metabolomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2037))

Abstract

Metabolomics represents a powerful, complementary approach for studying biological system responses to various biotic and abiotic stimuli. A major challenge in metabolomics is the lack of reliable annotations for all metabolites detected in complex MS and/or NMR data. To meet this challenge, we have developed an integrated UHPLC-QTOF-MS/MS-SPE-NMR system for higher-throughput metabolite identifications, which provides advanced biological context and enhances the scientific value of metabolomics data for understanding systems biology. This integrated instrumental method is less labor-intensive and more cost-effective than conventional individual methods (LC; MS; SPE; NMR). It enables the simultaneous purification and identification of primary and secondary metabolites present in biological samples. In this chapter, we describe the configuration and use of UHPLC-MS/MS-SPE-NMR in metabolite analyses ranging from sample extraction to higher-throughput metabolite annotation. With the integrated UHPLC-QTOF-MS/MS-SPE-NMR method, we have purified and confidently identified more than 100 previously known as well as unknown triterpene and flavonoid glycosides while noting that most of the identified compounds are not commercially available.

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Acknowledgments

The Sumner lab has been graciously supported by several entities over the years for the development of metabolite identifications, natural products profiling, and plant metabolomics. These specifically include support from the University of Missouri, the Samuel Roberts Noble Foundation, Bruker BioSpin and Daltonics Gmbh, Agilent Technologies, NSF-JST Metabolomics for a Low Carbon Society #1139489, NSF-MRI-DBI #1126719, NSF-RCN #1340058, and NSF-MCB #1024976. We thank Drs. Ulrich Braumann (Bruker BioSpin), Aiko Barsch (Bruker Daltonics), Daniel J. Wherritt (Noble Foundation), and Dennis Fine (Noble Foundation) for their operational and technical guidance in assembling the UHPLC-MS-SPE-NMR ensemble.

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Authors and Affiliations

  1. Department of Biochemistry, University of Missouri at Columbia, Columbia, MO, USA

    Anil Bhatia, Saurav J. Sarma, Zhentian Lei & Lloyd W. Sumner

  2. MU Metabolomics Center, University of Missouri at Columbia, Columbia, MO, USA

    Anil Bhatia, Saurav J. Sarma, Zhentian Lei & Lloyd W. Sumner

  3. Bond Life Sciences Center, University of Missouri at Columbia, Columbia, MO, USA

    Anil Bhatia, Saurav J. Sarma, Zhentian Lei & Lloyd W. Sumner

Authors
  1. Anil Bhatia
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  2. Saurav J. Sarma
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  3. Zhentian Lei
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  4. Lloyd W. Sumner
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Corresponding author

Correspondence to Lloyd W. Sumner .

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Editors and Affiliations

  1. Northwest Metabolomics Research Center, Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington, Seattle, WA, USA

    G. A. Nagana Gowda

  2. Northwest Metabolomics Research Center, Mitochondria and Metabolism Center, Department of Anesthesiology and Pain Medicine, University of Washington, Fred Hutch Cancer Research Center, Seattle, WA, USA

    Daniel Raftery

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Bhatia, A., Sarma, S.J., Lei, Z., Sumner, L.W. (2019). UHPLC-QTOF-MS/MS-SPE-NMR: A Solution to the Metabolomics Grand Challenge of Higher-Throughput, Confident Metabolite Identifications. In: Gowda, G., Raftery, D. (eds) NMR-Based Metabolomics. Methods in Molecular Biology, vol 2037. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9690-2_7

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  • DOI: https://doi.org/10.1007/978-1-4939-9690-2_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9689-6

  • Online ISBN: 978-1-4939-9690-2

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