Study of factors affecting the determination of total plasma 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD)-thiol derivatives by liquid chromatography

V. Rizzo, L. Montalbetti, M. Valli, T. Bosoni, E. Scoglio, R. Moratti

Research output: Contribution to journalArticle

Abstract

A detailed investigation of the factors affecting the determination of total plasma 7-fluorobenzo-2-oxa-1,3-diazole-4-sulfonate (SBD)-thiol derivatives (i.e. cysteine, homocysteine and cysteinylglycine) is described. Essentially, this assay entails extracting specific thiols by plasma disulphide bond reduction, protein precipitation, sulphydryl compound derivatization with the thiol-specific fluorogenic reagent ammonium 7-fluorobenzo-2-oxa-1,3-diazole-4-sulphonate (SBD-F), and subsequent separation with isocratic reversed-phase high-performance liquid chromatography. By improving the reliability of several analytical parameters (composition of the mobile phase, pretreatment of the sample using different reducing and protein precipitation agents, and optimization of the derivatization of thiols with SBD-F), a number of critical issues can be identified and solved.

Original languageEnglish
Pages (from-to)209-215
Number of pages7
JournalJournal of Chromatography B: Biomedical Applications
Volume706
Issue number2
DOIs
Publication statusPublished - Mar 20 1998

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Keywords

  • 7-Fluorobenzo-2-oxa-1,3-diazole-4-sulfonate
  • Amino acids
  • Cysteine
  • Cysteinylglycine
  • Derivatization
  • Homocysteine
  • LC
  • Thiols

ASJC Scopus subject areas

  • Chemistry(all)

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