Separation of 1-phenyl-3-methyl-5-pyrazolone derivatives of monosaccharides by capillary electrochromatography.
1-Phenyl-3-methyl-5-pyrazolone (PMP) derivatives of component monosaccharides in glycoproteins (fucose, galactose, mannose, N-acetylgalactosamine and N-acetylglucosamine) and epimeric aldopentoses (arabinose, lyxose, ribose and xylose) were well separated from each other by capillary electrochromatography on a Hypersil ODS column with a mixture of 50 mM N-(2-hydroxyethyl)piperazine-2'-(2-ethanesulfonic acid) buffer, pH 6.0 to approximately 6.3, and acetonitrile (2.2:1 v/v) as eluent. The elution of these compounds showed relatively strong dependence on the pH and concentration of the buffer salts contained in the eluent, as compared to the elution by pressure-driven high-performance liquid chromatography (HPLC) on the same stationary phase, but separation of PMP-monosaccharides was better than that by HPLC. Retention times of PMP-monosaccharides were highly reproducible with a relative standard deviation (RSD) of approximately 0.6%, and quantification with an RSD less than 5% could be achieved using 3-O-methylglucose as an internal standard.
Suzuki
S
S
Faculty of Pharmaceutical Sciences, Kinki University, Higashi-osaka, Japan.
Yamamoto
M
M
Kuwahara
Y
Y
Makiura
K
K
Honda
S
S
D000983:Antipyrine / Q000031:analogs & derivatives* / Q000302:isolation & purification; D002851:Chromatography, High Pressure Liquid; D000077553:Edaravone; D019075:Electrophoresis, Capillary / Q000379:methods; D009005:Monosaccharides / Q000302:isolation & purification*; D011720:Pyrazoles / Q000302:isolation & purification*; D015203:Reproducibility of Results
D009005:Monosaccharides; D011720:Pyrazoles; D000077553:Edaravone; D000983:Antipyrine