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Mass Spectrometry in Metabolomics pp 15–27Cite as

Global Metabolic Profiling Using Ultra-Performance Liquid Chromatography/Quadrupole Time-of-Flight Mass Spectrometry

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1198))

Abstract

Currently, liquid chromatography-mass spectrometry (LC-MS) is one of the most important analytical technologies for detecting hundreds of metabolites in the field of metabolomics. A recent advance in LC that has impacted metabolomics is the development of UPLC (ultra-performance liquid chromatography). In this chapter, we describe the analytical methodologies for the global metabolic profiling of serum, urine, and tissue samples using UPLC-Q-TOF (quadrupole-time-of-flight)-MS. Aqueous metabolites are extracted after adding methanol/acetonitrile/acetone and then analyzed by UPLC-MS under positive and/or negative ionization mode. With the aid of multivariate statistical analysis, separation between various groups can be observed in the score plots, and biomarkers are screened in the loading/weight/VIP (variable importance in the projection) scatterplots. Furthermore, putative markers can be identified through comparison with the authentic standards based on tandem mass spectrometry (MS/MS) fragmentation pattern and LC retention. We expect that our protocol, with modifications if necessary, can be useful in many metabolomics studies and a wide range of research areas related to small molecules and LC-MS.

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Acknowledgments

This work was supported by the National Science and Technology Major Project of the Ministry of Science and Technology of China (2013ZX09103-001), the National Natural Science Foundation of China (No. 21365001), the Chinese National Instrumentation Program (No. 2011YQ170067), and Li-Shi-Zhen Young Scientist Project of School of Pharmacy at the Second Military Medical University, Shanghai, China.

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Authors and Affiliations

  1. Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai, 200433, China

    Yunpeng Qi, Yunlong Song, Guorong Fan & Yifeng Chai

  2. Department of Anesthesiology & Pain Medicine, Northwest Metabolomics Research Center, University of Washington, Seattle, WA, 98195, USA

    Haiwei Gu

  3. Jiangxi Key Laboratory for Mass Spectrometry and Instrumentation, East China Institute of Technology, Nanchang, Jiangxi, China

    Haiwei Gu

Authors
  1. Yunpeng Qi
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  2. Yunlong Song
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  3. Haiwei Gu
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  4. Guorong Fan
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  5. Yifeng Chai
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Corresponding author

Correspondence to Yifeng Chai .

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Editors and Affiliations

  1. University of Washington Fred Hutchinson Cancer Rsrch Ctr, Seattle, Washington, USA

    Daniel Raftery

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Qi, Y., Song, Y., Gu, H., Fan, G., Chai, Y. (2014). Global Metabolic Profiling Using Ultra-Performance Liquid Chromatography/Quadrupole Time-of-Flight Mass Spectrometry. In: Raftery, D. (eds) Mass Spectrometry in Metabolomics. Methods in Molecular Biology, vol 1198. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1258-2_2

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  • DOI: https://doi.org/10.1007/978-1-4939-1258-2_2

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1257-5

  • Online ISBN: 978-1-4939-1258-2

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