Rapid analysis of amino sugars by microchip electrophoresis with laser-induced fluorescence detection.
Microchip electrophoresis for the short-time analysis of amino sugars is described. D-Glucosamine, D-galactosamine and their reduced forms were labeled with 4-nitro-2,1,3-benzoxadiazole 7-fluoride (NBD-F) at pH 6.0 and the fluorescent derivatives were purified on an octadecyl silica (ODS) gel plate. The derivatives were analyzed by electrophoresis on a microfabricated chip with a 33 mm long separation channel with argon laser-induced fluorescence detection. Under the established conditions, these amino sugarderivatives were well separated from each other within 60 s. Amino sugars of as small an amount as 0.5 fmol could be detected with a signal-to-noise (S/N) ratio of 3, and peak response showed good linearity between at least 0.8 and 8 fmol of samples with a relative standard deviation (RSD) of ca. 4%. This method was also applied to the analysis of amino sugar composition of O-linked glycans released from bovine submaxillary mucin with alkali in the presence of borohydride. The result of amino sugar composition analysis for individual O-glycans fractionated by high-performance liquid chromatography was quite useful for their identification.
Suzuki
S
S
Faculty of Pharmaceutical Sciences, Kini, University, Higashi-osaka, Japan
Shimotsu
N
N
Honda
S
S
Arai
A
A
Nakanishi
H
H
D009327:4-Chloro-7-nitrobenzofurazan / Q000031:analogs & derivatives* / Q000032:analysis; D000606:Amino Sugars / Q000032:analysis*; D000818:Animals; D001894:Borohydrides; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid; D002855:Chromatography, Thin Layer; D019075:Electrophoresis, Capillary / Q000295:instrumentation*; D004867:Equipment Design; D005456:Fluorescent Dyes / Q000032:analysis; D005470:Fluorometry / Q000295:instrumentation*; D005688:Galactosamine / Q000032:analysis; D005944:Glucosamine / Q000032:analysis; D006863:Hydrogen-Ion Concentration; D007834:Lasers; D008832:Microchemistry / Q000295:instrumentation*; D008969:Molecular Sequence Data; D009077:Mucins / Q000737:chemistry / Q000302:isolation & purification; D009844:Oligosaccharides / Q000032:analysis; D010084:Oxidation-Reduction; D011134:Polysaccharides / Q000737:chemistry; D012680:Sensitivity and Specificity; D013363:Submandibular Gland / Q000378:metabolism
D000606:Amino Sugars; D001894:Borohydrides; D005456:Fluorescent Dyes; D009077:Mucins; D009844:Oligosaccharides; D011134:Polysaccharides; C032930:7-fluoro-4-nitrobenzo-2-oxa-1,3-diazole; D005688:Galactosamine; D009327:4-Chloro-7-nitrobenzofurazan; D005944:Glucosamine