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A Sensitive Derivatization Method for the Determination of the Sugar Composition after Pre‐column Reductive Amination with 3‐Amino‐9‐ethylcarbazole (AEC) by High‐Performance Liquid Chromatography
Author(s) -
ZHANG Ying,
HUANG LinJuan,
WANG ZhongFu
Publication year - 2007
Publication title -
chinese journal of chemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.28
H-Index - 41
eISSN - 1614-7065
pISSN - 1001-604X
DOI - 10.1002/cjoc.200790280
Subject(s) - chemistry , derivatization , monosaccharide , chromatography , high performance liquid chromatography , xylose , rhamnose , arabinose , detection limit , reductive amination , polysaccharide , organic chemistry , catalysis , fermentation
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 °C 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.