Rhodamine B amine as a highly sensitive fluorescence derivatization reagent for saccharides in reversed-phase liquid chromatography.
6-Rhodamine B amine functions as a highly sensitive fluorescence derivatization reagent for mono- and oligosaccharides; it reacts with the reducing end of saccharides under acidic conditions. The fluorescent derivatives of five monosaccharides can be separated within 25 min by reversed-phase liquid chromatography with isocratic elution. The detection limits (S/N = 3) for mono-, di-, and oligosaccharides are 7-51, 13, and 9-35 fmol/20 microl injection, which correspond to analyte concentrations of 35-255, 65, 45-175 nM, respectively. We have applied this derivatization method successfully to the analysis of the components of oligosaccharides in glycoproteins (ribonuclease B and fetuin) following their acidic or enzymatic hydrolysis. The results from these analyses are in good agreements with the reported values established previously.
Todoroki
Kenichiro
K
Faculty of Pharmaceutical Sciences, Fukuoka University, 8-19-1 Nanakuma, Johnan-ku, Fukuoka 814-0180, Japan.
Hayama
Tadashi
T
Ijiri
Soichiro
S
Kazuta
Asako
A
Yoshida
Hideyuki
H
Nohta
Hitoshi
H
Yamaguchi
Masatoshi
M
D002240:Carbohydrate Sequence; D002241:Carbohydrates / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D008969:Molecular Sequence Data; D012235:Rhodamines / Q000737:chemistry*
D002241:Carbohydrates; D005456:Fluorescent Dyes; D012235:Rhodamines; C029773:rhodamine B