Analysis of sucrose acetates in a crude 6-O-acetyl sucrose product by on-line hydrolysis-high-performance liquid chromatography with pulsed amperometric detection

A standard-free and sensitive method was developed for analysis of sucrose acetates in a crude 6- O -acetyl sucrose (S-6-a) product by on-line hydrolysis-high-performance liquid chromatography with pulsed amperometric detection (PAD). Sucrose, three regio-isomers of acetyl sucrose and five regio-isomers of diacetyl sucrose were separated on a C18 column using 3% (v/v) acetonitrile in water as eluent within 25   min. After purification with LC followed by semi-preparative HPLC, their chemical structures were identified by 1D, 2D NMR and LC–MS. Moreover, quantification of those regio-isomers was achieved by on-line alkaline hydrolysis to liberate sucrose using a post-column delivery system, and then detected by PAD for indirect estimation of the sucrose acetate content. Under optimal conditions, the linear ranges were from 0.03 to 150   μmol   L −1 for sucrose corresponding to sucrose acetates with coefficient of determination as 0.9997 and detection limit as 0.01   μmol   L −1 ( S/N   =   3). Good repeatability was obtained (RSD   <   3%, n   =   6). Furthermore, this method has been successfully applied to the analysis of sucrose and sucrose acetates in a crude S-6-a product during synthesis, purification and structure elucidation studies. The recoveries were from 94.89% to 102.31% for sucrose and sucrose acetates.