Determination of selenomethionine by high-performance liquid chromatography-fluorescence detection coupled with on-line oxidation.
A simple and sensitive determination method for selenomethionine (Se-Met) using an HPLC-fluorescence detection system coupled with an on-line electrochemical reactor has been developed. NBD-F (4-fluoro-7-nitro-2,1,3-benzoxadiazole) was used as the fluorescent derivatization reagent for Se-Met. NBD-Se-Met was separated from NBD-derivatives of 22 other amino acids within 35 min. Applying an optimized oxidation potential enhanced the fluorescence intensity of NBD-Se-Met 10-fold. The calibration curve was linear in the range of 300 fmol to 30 pmol with a correlation coefficient of 0.997. Detection limit (S/N = 3) was calculated to be 50 fmol, which is comparable to that of inductively coupled plasma mass spectrometry. This simple and sensitive method should be useful for the determination of Se-Met in physiological samples, such as serum or urine.
Aoyama
Chiaki
C
Graduate School of Pharmaceutical Sciences, The University of Tokyo, Bunkyo, Tokyo 113-0033, Japan.
Tsunoda
Makoto
M
Funatsu
Takashi
T
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives; D002138:Calibration; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005453:Fluorescence; D010084:Oxidation-Reduction; D012645:Selenomethionine / Q000032:analysis*
C032930:7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole; D012645:Selenomethionine; D009327:4-Chloro-7-nitrobenzofurazan