Bromide-sulfur interchange: Ion chromatographic determination of total reduced thiol levels in plasma

SA Chitre, GAM Lobo, SM Rathod, RB Smith, R Leslie, C Livingstone, J Davis

    Research output: Contribution to journalArticle

    18 Citations (Scopus)

    Abstract

    Plasma thiol concentration has long been recognised as a potential indicator for assessing the severity of oxidative stress processes within physiological systems. While such measurements are normally restricted to research studies, this communication has sought to develop and characterise a novel approach through which this parameter could be exploited within routine clinical settings. The protocol is based on the rapid derivatisation of reduced thiol functionalities (protein and monomolecular moieties) through the homogenous reaction of a naphthoquinone bromide derivative. Bromide released in the reaction can be easily quantified through ion chromatography (Isocractic Dionex DX-120 incorporating an lonPac (R) AS 14 anion exchange column and a 25 mu L sample loop with conductivity detector. Mobile phase consisted sodium carbonate/bicarbonate (3.5 mM/1 mM) at a flow rate of 1.5 mL/min). Method selectivity and sensitivity has been critically evaluated. The technique covers the range 15 mu M-3.5 mM PSH with a detection limit of 9 mu M PSH and analysis time of 5 min. The efficacy of the approach for the analysis of human plasma from five volunteers was assessed (ranging from 49 to 72 mu M with an intra assay variation of less than 5% in all cases). The responses were validated through comparison with the standard Ellman colorimetric technique. (C) 2008 Elsevier B.V. All rights reserved.
    Original languageEnglish
    Pages (from-to)173-177
    JournalJournal of Chromatography B
    Volume864
    Issue number1-2
    DOIs
    Publication statusPublished - Mar 2008

      Fingerprint

    Keywords

    • albumin
    • plasma
    • thiol
    • bromide
    • ion chromatography
    • oxidative stress

    Cite this