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An approach for the development and selection of chromatographic methods for high-throughput metabolomic screening of urine by ultra pressure LC-ESI-ToF-MS

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Abstract

Since the components of a sample for open metabolomic analysis are unknown a priori a pragmatic approach to method development has been taken in order to develop and select a chromatographic method suitable for high-throughput open metabolomic screening of urine by Ultra Performance Liquid Chromatography-Mass Spectrometry (UPLC-MS). A total of 848 injections of diluted rat urine were made onto a UPLC-ESI-ToF-MS system using several different gradient profiles and run times to determine a suitable method for analysis of urine from male and female rats. Peak integral and multivariate data analysis were performed to investigate the quality of separation and information obtained from these multiple analyses. A suitable 8 min method was selected and is now used routinely for open profiling metabolomic analyses of urine. The use of a sample-relevant QC mix is also discussed.

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Acknowledgements

The authors would like to thank Rachel Ball (GSK) for her assistance with UPLC-MS data acquisition, Brian Sweatman and Susan Connor (GSK) for advice regarding the composition of Artificial Urine, and Dan McMillan and Hilary Major (Waters) for advice relating to OpenLynx and MarkerLynx software respectively. M.P·H. would like to thank GlaxoSmithKline for the financial support of his Ph.D. programme. J.L.G. is a Royal Society University Research Fellow.

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Authors and Affiliations

  1. Safety Assessment Division, GlaxoSmithKline, Park Road, Ware, Hertfordshire, SG12 0DP, UK

    Mark P. Hodson & John N. Haselden

  2. Department of Biochemistry, University of Cambridge, Tennis Court Road, Cambridge, CB2 1QW, UK

    Mark P. Hodson & Julian L. Griffin

  3. DMPK Division, GlaxoSmithKline, Park Road, Ware, Hertfordshire, SG12 0DP, UK

    Gordon J. Dear

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  1. Mark P. Hodson
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Correspondence to Mark P. Hodson.

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Hodson, M.P., Dear, G.J., Griffin, J.L. et al. An approach for the development and selection of chromatographic methods for high-throughput metabolomic screening of urine by ultra pressure LC-ESI-ToF-MS. Metabolomics 5, 166–182 (2009). https://doi.org/10.1007/s11306-008-0135-1

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