3-Amino-9-ethylcarbazole (AEC) was employed for monosaccharide derivatization. The derivatives can be analyzed by high-performance liquid chromatography (HPLC) with an ultraviolet detection (wavelength: 254 nm)...3-Amino-9-ethylcarbazole (AEC) was employed for monosaccharide derivatization. The derivatives can be analyzed by high-performance liquid chromatography (HPLC) with an ultraviolet detection (wavelength: 254 nm). Monosaccharides were quantitatively derivatized with 3-amino-9-ethylcarbazole (AEC) at 70 ℃ for 60 min. The method was linear for all samples over the concentration range tested (r〉0.999), the precision was found to be satisfactory (R.S.D. 〈 3%), and the recovery ratios were 〉98.62%. The stability analysis showed R.S.D. was between 1.81%-3.16%. Detection limits for the samples (D-glucose, L-xylose, D-mannose, L-arabinose, and L-rhamnose) ranged from 0.06 to 1.97 ng/mL (S/N=3). Under the optimized derivatization and HPLC conditions, five monosaccharides were well separated using a narrow bore C18 column (250 mm×4.6 mm) with 0.1 mol/L ammonium acetate containing 25% acetonitrile at a flow rate of 0.5 mL/min. As an application, the method has been successfully applied to the determination of monosaccharide compositions of three polysaccharides SPPA-1, SPPB-1 and SPPC- 1 of Spirulina platensis. This method also has potential application to oligosaccharide or glycan analyses.展开更多
基金Project supported by the Hi-Tech Research and Development program of China (No. 2006AA02Z146), the Natural Science Foundation of Shaanxi Province (No. 2005C131) and the Science and Technology Key Program of Ministry of Education of China (No. 206147).
文摘3-Amino-9-ethylcarbazole (AEC) was employed for monosaccharide derivatization. The derivatives can be analyzed by high-performance liquid chromatography (HPLC) with an ultraviolet detection (wavelength: 254 nm). Monosaccharides were quantitatively derivatized with 3-amino-9-ethylcarbazole (AEC) at 70 ℃ for 60 min. The method was linear for all samples over the concentration range tested (r〉0.999), the precision was found to be satisfactory (R.S.D. 〈 3%), and the recovery ratios were 〉98.62%. The stability analysis showed R.S.D. was between 1.81%-3.16%. Detection limits for the samples (D-glucose, L-xylose, D-mannose, L-arabinose, and L-rhamnose) ranged from 0.06 to 1.97 ng/mL (S/N=3). Under the optimized derivatization and HPLC conditions, five monosaccharides were well separated using a narrow bore C18 column (250 mm×4.6 mm) with 0.1 mol/L ammonium acetate containing 25% acetonitrile at a flow rate of 0.5 mL/min. As an application, the method has been successfully applied to the determination of monosaccharide compositions of three polysaccharides SPPA-1, SPPB-1 and SPPC- 1 of Spirulina platensis. This method also has potential application to oligosaccharide or glycan analyses.