High-performance liquid chromatography of reducing carbohydrates as strongly ultraviolet-absorbing and electrochemically sensitive 1-phenyl-3-methyl-5-pyrazolone derivatives.
We found that 1-phenyl-3-methyl-5-pyrazolone reacts with reducing carbohydrates almost quantitatively to yield 2:1 compounds having no stereoisomers, which strongly absorb the uv light at 245 nm and are easily oxidizable on a glassy carbon electrode. Reverse-phase partition chromatography on a column of Capcell Pak C18 with uv or electrochemical detection allowed rapid analysis of aldoses and N-acetylhexosamines with the detection limit of 1 pmol or 100 fmol, respectively. This method proved especially useful for analysis of component monosaccharides of glycorproteins. It was also shown to be valid for separation of reducing oligosaccharides; maltodextrins with a degree of polymerization up to 19 were similarly derivatized and separated on this stationary phase.
Honda
S
S
Faculty of Pharmaceutical Sciences, Kinki University, Higashi-osaka, Japan.
Akao
E
E
Suzuki
S
S
Okuda
M
M
Kakehi
K
K
Nakamura
J
J
D000818:Animals; D000983:Antipyrine / Q000031:analogs & derivatives*; D002241:Carbohydrates / Q000032:analysis*; D002247:Carbon Isotopes; D002417:Cattle; D002645:Chickens; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D000077553:Edaravone; D004563:Electrochemistry; D005260:Female; D005947:Glucose / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D009682:Magnetic Resonance Spectroscopy; D009005:Monosaccharides / Q000032:analysis; D009844:Oligosaccharides / Q000032:analysis; D012015:Reference Standards; D015203:Reproducibility of Results; D013056:Spectrophotometry, Ultraviolet; D013552:Swine
D002241:Carbohydrates; D002247:Carbon Isotopes; D006023:Glycoproteins; D009005:Monosaccharides; D009844:Oligosaccharides; D005947:Glucose; D000077553:Edaravone; D000983:Antipyrine