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Comparative evaluation of extraction methods for simultaneous mass-spectrometric analysis of complex lipids and primary metabolites from human blood plasma

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Abstract

Metabolomic results on human blood plasma largely depend on the sample preparation protocols employed for protein precipitation and metabolite extraction. Five different extraction methods were examined, which can be grouped into two categories, liquid-liquid extraction and protein precipitation methods, including long-standing protocols such as the Folch extraction and Bligh-Dyer extraction in comparison to modern methods such as the Matyash protocol and two global metabolite extraction methods. Extracts were subjected to analysis of blood plasma lipids and primary metabolites by using chip-based direct infusion nanoelectrospray tandem mass spectrometry and gas chromatography coupled to time-of-flight mass spectrometry, respectively. Optimal extraction schemes were evaluated based on the number of identified metabolites, extraction efficiency, compound diversity, reproducibility, and convenience for high-throughput sample preparations. Results showed that Folch and Matyash methods were equally valid and robust for lipidomic assessments while primary metabolites were better assessed by the protein precipitation methods with organic solvent mixtures.

Schematic workflow of five extraction methods and subsequent mass spectrometry analysis using GC-TOF MS and nanoelectrospray direct-infusion ion trap MS/MSᅟ

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Acknowledgments

This study was supported by the National Institutes of Health grants 1U24 DK097154 and P20 HL113452 to OF, the Bio & Medical Technology Development Program of the National Research Foundation (NRF) funded by the Ministry of Science, ICT & Future Planning (grant no. NRF-2013M3A9B6046416), the Korea Healthcare Technology R&D Project funded by Ministry of Health & Welfare (grant no. HN13C0076), and the Advanced Biomass R&D Center of Korea funded by the Korean Government (MSIP, grant no. 2011-0031353), Republic of Korea.

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Authors and Affiliations

  1. The Department of Bio and Fermentation Convergence Technology, Kookmin University, Seoul, 136-702, South Korea

    Do Yup Lee

  2. UC Davis Genome Center, Metabolomics, UC Davis, 451 Health Sciences Drive, Davis, CA, 95616, USA

    Do Yup Lee, Tobias Kind, Oliver Fiehn & Kwang-Hyeon Liu

  3. Department of Biomedical Science and Clinical Trial Center, BK21 PLUS KNU Bio-Medical Convergence Program for Creative Talent, Kyungpook National University Graduates School and Hospital, Daegu, 700-734, South Korea

    Young-Ran Yoon

  4. College of Pharmacy and Research Institute of Pharmaceutical Sciences, Kyungpook National University, Daegu, 702-701, South Korea

    Kwang-Hyeon Liu

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  1. Do Yup Lee
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  3. Young-Ran Yoon
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Correspondence to Oliver Fiehn or Kwang-Hyeon Liu.

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Lee, D.Y., Kind, T., Yoon, YR. et al. Comparative evaluation of extraction methods for simultaneous mass-spectrometric analysis of complex lipids and primary metabolites from human blood plasma. Anal Bioanal Chem 406, 7275–7286 (2014). https://doi.org/10.1007/s00216-014-8124-x

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  • DOI: https://doi.org/10.1007/s00216-014-8124-x

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