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Pre-analytic evaluation of volumetric absorptive microsampling and integration in a mass spectrometry-based metabolomics workflow

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Abstract

Volumetric absorptive microsampling (VAMS) is a novel approach that allows single-drop (10 μL) blood collection. Integration of VAMS with mass spectrometry (MS)-based untargeted metabolomics is an attractive solution for both human and animal studies. However, to boost the use of VAMS in metabolomics, key pre-analytical questions need to be addressed. Therefore, in this work, we integrated VAMS in a MS-based untargeted metabolomics workflow and investigated pre-analytical strategies such as sample extraction procedures and metabolome stability at different storage conditions. We first evaluated the best extraction procedure for the polar metabolome and found that the highest number and amount of metabolites were recovered upon extraction with acetonitrile/water (70:30). In contrast, basic conditions (pH 9) resulted in divergent metabolite profiles mainly resulting from the extraction of intracellular metabolites originating from red blood cells. In addition, the prolonged storage of blood samples at room temperature caused significant changes in metabolome composition, but once the VAMS devices were stored at − 80 °C, the metabolome remained stable for up to 6 months. The time used for drying the sample did also affect the metabolome. In fact, some metabolites were rapidly degraded or accumulated in the sample during the first 48 h at room temperature, indicating that a longer drying step will significantly change the concentration in the sample.

Volumetric absorptive microsampling (VAMS) is a novel technology that allows single-drop blood collection and, in combination with mass spectrometry (MS)-based untargeted metabolomics, represents an attractive solution for both human and animal studies. In this work, we integrated VAMS in a MS-based untargeted metabolomics workflow and investigated pre-analytical strategies such as sample extraction procedures and metabolome stability at different storage conditions. The latter revealed that prolonged storage of blood samples at room temperature caused significant changes in metabolome composition, but if VAMS devices were stored at − 80 °C, the metabolome remained stable for up to 6 months

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Acknowledgements

This work was supported by the FWF-funded doctoral program HOROS and by a transnational PhD research project between the Medical University of Innsbruck and the EURAC research in Bolzano (BI-DOC project).

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Authors and Affiliations

  1. Institute for Biomedicine, Eurac Research, Affiliated Institute of the University of Lübeck, Via Galvani 31, 39100, Bolzano/Bozen, Italy

    Chiara Volani, Giulia Caprioli, Giovanni Calderisi, Baldur B. Sigurdsson, Johannes Rainer, Andrew A. Hicks, Peter P. Pramstaller, Sigurdur V. Smarason & Giuseppe Paglia

  2. Department of Internal Medicine II (Infectious Diseases, Immunology, Rheumatology and Pneumology), Medical University of Innsbruck, Anichstrasse 35, 6020, Innsbruck, Austria

    Chiara Volani & Guenter Weiss

  3. Transfusion Center of the Hospital of Bolzano, Lorenz Böhler Str. 5, 39100, Bozen, Italy

    Ivo Gentilini

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  1. Chiara Volani
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Correspondence to Giuseppe Paglia.

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This study was performed in accordance with the ethical standards. The local ethics committee (Comitato etico del comprensorio sanitario di Bolzano) approved the study, and all participants provided written informed consent.

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Volani, C., Caprioli, G., Calderisi, G. et al. Pre-analytic evaluation of volumetric absorptive microsampling and integration in a mass spectrometry-based metabolomics workflow. Anal Bioanal Chem 409, 6263–6276 (2017). https://doi.org/10.1007/s00216-017-0571-8

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