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.

Salvatore Sotgia, Elisabetta Pisanu, Gianfranco Pintus, Gian Luca Erre, Gerard Aime Pinna, Luca Deiana, Ciriaco Carru, Angelo Zinellu

PloS one 2013

filter terms:

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%).

Citation

Salvatore Sotgia, Elisabetta Pisanu, Gianfranco Pintus, Gian Luca Erre, Gerard Aime Pinna, Luca Deiana, Ciriaco Carru, Angelo Zinellu. 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;8(7):e70374