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Urine stability for metabolomic studies: effects of preparation and storage

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Metabolomic studies attempt to identify and profile unique metabolic differences among test populations, which may be correlated with a specific biological stress or pathophysiology. Due to the ease of collection and the metabolite-rich nature of urine, it is frequently used as a bio-fluid for human and animal metabolic studies. High-resolution 1H-NMR is an analytical tool used to qualitatively and quantitatively identify metabolites in urine. Urine samples were collected from healthy male and female subjects and prepared: raw, following centrifugation, filtration, or the addition of the bacteriostatic preservative sodium azide and analyzed by NMR. In addition, these samples were stored at room temperature (22 °C), in a refrigerator (4 °C), or in a deep-freeze (−80 °C). Samples were analyzed by NMR every week for a month and changes in concentrations of 55 easily identifiable metabolites were followed. The degree of change in metabolite concentrations following storage over a 4-week period were influenced by the different methods of sample preparation and storage. Significant changes in urine metabolites are likely due to bacterial contamination of the urine. Our study demonstrates that bacterial contamination of urine in normal individuals significantly alters the metabolic profile of urine over time and proper preparation and storage procedures must be followed to reduce these changes. By identifying appropriate methods of urine preparation and storage investigators will preserve the fidelity of the urine samples in order to better reflect the original metabolic state.

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Acknowledgments

This research was supported by the Canadian Institutes of Health Research, Genome Prairie, Genome Canada, the Natural Science and Engineering Research Council of Canada, and the University of Alberta.

The authors would like to thank Angela Thiessen for her assistance in sample preparation. We would also like to thank Carolyn Slupsky for her helpful discussions. This research was supported by the Canadian Institues of Health Research (CIHR), Genome Prairie, and Genome Canada. We would like to thank the Canadian National High Field NMR Centre (NANUC) for their assistance and use of the facilities. Operation of NANUC is funded by CIHR, the Natural Science and Engineering Research Council of Canada and the University of Alberta.

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Authors and Affiliations

  1. CIHR Group in Protein Structure and Function, University of Alberta, Edmonton, Alberta, Canada

    Erik J. Saude & Brian D. Sykes

  2. Department of Biochemistry, University of Alberta, 4-19 Medical Sciences Building, Edmonton, Alberta, Canada, T6G 2E1

    Erik J. Saude & Brian D. Sykes

  3. Medical Resonance Diagnostics Centre (MDRC), University of Alberta, Edmonton, Alberta, Canada

    Erik J. Saude & Brian D. Sykes

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  1. Erik J. Saude
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Correspondence to Brian D. Sykes.

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Saude, E.J., Sykes, B.D. Urine stability for metabolomic studies: effects of preparation and storage. Metabolomics 3, 19–27 (2007). https://doi.org/10.1007/s11306-006-0042-2

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