Plasma L-ergothioneine measurement by high-performance liquid chromatography and capillary electrophoresis after a pre-column derivatization with 5-iodoacetamidofluorescein (5-IAF) and fluorescence detection

PLoS One. 2013 Jul 29;8(7):e70374. doi: 10.1371/journal.pone.0070374. Print 2013.

Abstract

Two sensitive and reproducible capillary electrophoresis and high-performance liquid chromatography-fluorescence procedures were established for quantitative determination of L-egothioneine in plasma. After derivatization of L-ergothioneine with 5-iodoacetamidofluorescein, the separation was carried out by HPLC on an ODS-2 C-18 sperisorb column by using a linear gradient elution and by HPCE on an uncoated fused silica capillary, 50 µm id, and 60 cm length. The methods were validated and found to be linear in the range of 0.3 to 10 µmol/l. The limit of quantification was 0.27 µmol/l for HPCE and 0.15 µmol/l for HPLC. The variations for intra- and inter-assay precision were around 6 RSD%, and the mean recovery accuracy close to 100% (96.11%).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Chromatography, High Pressure Liquid
  • Electrophoresis, Capillary
  • Ergothioneine / blood*
  • Ergothioneine / chemistry
  • Fluoresceins / chemistry
  • Humans
  • Reproducibility of Results
  • Sensitivity and Specificity
  • Spectrometry, Fluorescence

Substances

  • Fluoresceins
  • 5-iodoacetamidofluorescein
  • Ergothioneine

Grants and funding

Funding was provided by the “Fondazione Banco di Sardegna”, Sassari, Italy and by the “Ministero dell’Università e della Ricerca”, Italy. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.