title: Characterization of saccharide using high fluorescent 5-(((2-(carbohydrazino)methyl)thio)acetyl)-aminofluorescein tag by Capillary-HPLC-LIF and MALDI-TOF-MS.
abstract: The new approach to one-step derivatization of saccharide with 5-(((2-(carbohydrazino)methyl)thio)acetyl)-aminofluorescein (C356) was described. In this approach, high fluorescent C356 was applied to label saccharide to enhance the response of derivative saccharide and high sensitive capillary high performance liquid chromatography with laser-induced fluorescence (Capillary-HPLC-LIF) associated with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was used to characterize C356 labeled saccharide. The effect of derivatization conditions was evaluated and discussed. The limit of detection (LOD) of neutral saccharide in our method attained the level of femtomolar. As a result, this method could be successfully applied to determine the structure of N-glycans of glycoprotein.
authors.affiliation: Department of Chemistry & Institutes of Biomedical Sciences, Fudan University, Shanghai 200433, PR China.
authors.forename: Chaofeng
authors.initials: C
authors.lastname: Wang
authors.forename: Mingxia
authors.initials: M
authors.lastname: Gao
authors.forename: Zhi
authors.initials: Z
authors.lastname: Huang
authors.forename: Xiangmin
authors.initials: X
authors.lastname: Zhang
mesh_terms: D019342:Acetic Acid / Q000737:chemistry; D000818:Animals; D002240:Carbohydrate Sequence; D002417:Cattle; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005452:Fluoresceins / Q000737:chemistry*; D005936:Glucans / Q000032:analysis* / Q000737:chemistry; D006834:Hydrazines / Q000737:chemistry*; D006868:Hydrolysis; D057230:Limit of Detection; D008969:Molecular Sequence Data; D012259:Ribonuclease, Pancreatic / Q000737:chemistry*; D013050:Spectrometry, Fluorescence / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: C121151:5-(((2-(carbohydrazino)methyl)thio)acetyl)aminofluorescein aminooxyacetyl hydrazide; D005452:Fluoresceins; D005936:Glucans; D006834:Hydrazines; C031146:maltoheptaose; D012259:Ribonuclease, Pancreatic; C504335:pancreatic ribonuclease B, bovine; D019342:Acetic Acid
keywords: C356; Capillary-HPLC-LIF; Derivatization; Saccharide