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.
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%).
Department of Biomedical Sciences, University of Sassari, Sassari, Italy. salvatore.sotgia@gmail.com
Salvatore
S
Sotgia
Elisabetta
E
Pisanu
Gianfranco
G
Pintus
Gian Luca
GL
Erre
Gerard Aime
GA
Pinna
Luca
L
Deiana
Ciriaco
C
Carru
Angelo
A
Zinellu
D002851:Chromatography, High Pressure Liquid; D019075:Electrophoresis, Capillary; D004880:Ergothioneine / Q000097:blood* / Q000737:chemistry; D005452:Fluoresceins / Q000737:chemistry; D006801:Humans; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
D005452:Fluoresceins; C015296:5-iodoacetamidofluorescein; D004880:Ergothioneine