Cleanup and analysis of sugar phosphates in biological extracts by using solid phase extraction and anion-exchange chromatography with pulsed amperometric detection

Hans Peter Smith, A. Cohen, T. Buttler, Jens Bredal Nielsen, Lisbeth Olsson

    Research output: Contribution to journalJournal articleResearchpeer-review

    Abstract

    A cleanup method based on anion-exchange solid-phase extraction (SPE) was developed to render biological extracts suitable for the analysis of hexose phosphates with a modified anion-exchange chromatography method and pulsed amperometric detection. The method was applied to cell extracts of Saccharomyces cerevisiae obtained by using cold methanol as quenching agent and chloroform as extraction solvent. It was shown that pretreatment of the cell extract with SPE markedly improved the quality of the liquid chromatography analysis with recoveries of the sugar phosphates close to 100%. Furthermore, the method allowed for sample enrichment and the original extraction procedure could be simplified by implementing SPE early in the extraction protocol. (C) 1998 Academic Press.
    Original languageEnglish
    JournalAnalytical Biochemistry
    Volume261
    Pages (from-to)36-42
    ISSN0003-2697
    DOIs
    Publication statusPublished - 1998

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