Separation of human fibrinopeptides by capillary zone electrophoresis

Aldo Profumo, Barbara Cardinali, Carla Cuniberti, Mattia Rocco

Research output: Contribution to journalArticle

Abstract

We describe a method for the simultaneous determination of the five fibrinopeptide forms derived from the thrombin-promoted activation of human fibrinogen by capillary zone electrophoresis (CZE). The fibrinopeptide mixture was first desalted by a solid-phase extraction (SPE) step. The analysis was performed in reversed polarity in a highly cross-linked polyethylene glycol (PEG)-coated capillary with UV-light absorption detection at 200 nm. Several parameters including buffer concentration and pH, presence of an organic modifier, temperature, and applied voltage, have been tested. The best separations were obtained within 20 min, utilizing a 20 mM sodium phosphate buffer without organic modifier, in the narrow 6.1-6.2 pH range, at 25°C, with an applied voltage of 20 kV. Quantitative analysis is made possible by the use of sheep fibrinopeptide A as an internal standard to correct for both extraction and injection errors.

Original languageEnglish
Pages (from-to)600-609
Number of pages10
JournalElectrophoresis
Volume26
Issue number3
DOIs
Publication statusPublished - Feb 2005

Keywords

  • Blood coagulation
  • Capillary electrophoresis
  • Fibrinogen
  • Fibrinopeptides
  • Solid-phase extraction

ASJC Scopus subject areas

  • Clinical Biochemistry

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