title: A novel condition for capillary electrophoretic analysis of reductively aminated saccharides without removal of excess reagents.
abstract: We have identified novel CE conditions for the separation of 7-amino-4-methylcoumarin-labeled monosaccharides and oligosaccharides from glycoproteins. Using a neutrally coated capillary and alkaline borate buffer containing hydroxypropylcellulose and ACN, saccharide derivatives form anionic borate complexes, which move from the cathode to the anode in an electric field and are detected near the anodic end. Excess labeling reagents and other fluorescent products remain at the cathodic end. Fluorimetric detection using an LED as a light source enables determination of monosaccharide derivatives with good linearity between at least 0.4 and 400 μM, may correspond to 140 amol to 140 fmol. The lower LOD (S/N = 5) is only 80 nM in the sample solution (ca. 28 amol). The results were comparable to reported values using fluorometric detection LC. The method was also applied to the analysis of oligosaccharides that were enzymatically released from glycoproteins. Fine resolution enables profiling of glycans in glycoproteins. The applicability of the method was examined by applying it to other derivatives labeled with nonacidic tags such as ethyl p-aminobenzoate- and 2-aminoacridone-labeled saccharides.
authors.affiliation: Faculty of Pharmaceutical Sciences, Kinki University, Kowakae, Higashi-Osaka, Osaka, Japan.
authors.forename: Masahiro
authors.initials: M
authors.lastname: Yodoshi
authors.forename: Natsumi
authors.initials: N
authors.lastname: Ikeda
authors.forename: Naoko
authors.initials: N
authors.lastname: Yamaguchi
authors.forename: Mana
authors.initials: M
authors.lastname: Nagata
authors.forename: Noriaki
authors.initials: N
authors.lastname: Nishida
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Kakehi
authors.forename: Takao
authors.initials: T
authors.lastname: Hayakawa
authors.forename: Shigeo
authors.initials: S
authors.lastname: Suzuki
mesh_terms: D000818:Animals; D002417:Cattle; D002645:Chickens; D003374:Coumarins / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D006023:Glycoproteins / Q000737:chemistry*; D006801:Humans; D057230:Limit of Detection; D009005:Monosaccharides / Q000032:analysis* / Q000737:chemistry; D009844:Oligosaccharides / Q000032:analysis* / Q000737:chemistry
chemical_list: D003374:Coumarins; D006023:Glycoproteins; D009005:Monosaccharides; D009844:Oligosaccharides; C028743:7-amino-4-methylcoumarin
keywords: 7-Amino-4-methylcoumarin; Borate complex; Glycoprotein glycans; Monosaccharide analysis; Reductive amination