title: Analysis of sucrose acetates in a crude 6-O-acetyl sucrose product by on-line hydrolysis-high-performance liquid chromatography with pulsed amperometric detection.
abstract: 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 25min. 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 150μmolL(-1) for sucrose corresponding to sucrose acetates with coefficient of determination as 0.9997 and detection limit as 0.01μmolL(-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.
authors.affiliation: Department of chemistry, Zhejiang University, Hangzhou 310028, China.
authors.forename: Wenwu
authors.initials: W
authors.lastname: Yan
authors.affiliation: Department of Medicine, Zhejiang Academy of Traditional Chinese Medicine, Hangzhou, 310028, China.
authors.forename: Nani
authors.initials: N
authors.lastname: Wang
authors.affiliation: Department of chemistry, Zhejiang University, Hangzhou 310028, China.
authors.forename: Peimin
authors.initials: P
authors.lastname: Zhang
authors.affiliation: Department of chemistry, Zhejiang University, Hangzhou 310028, China.
authors.forename: Jiajie
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Zhejiang Institute of geology and mineral resources, Hangzhou 310007, China.
authors.forename: Shuchao
authors.initials: S
authors.lastname: Wu
authors.affiliation: Department of chemistry, Zhejiang University, Hangzhou 310028, China. Electronic address: zhuyan@zju.edu.cn.
authors.forename: Yan
authors.initials: Y
authors.lastname: Zhu
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods; D006868:Hydrolysis; D057230:Limit of Detection; D013237:Stereoisomerism; D013395:Sucrose / Q000031:analogs & derivatives* / Q000032:analysis / Q000737:chemistry
chemical_list: C556997:sucrose-6-acetate; D013395:Sucrose
keywords: 6-O-acetyl sucrose; HPLC; On-line hydrolysis; Pulsed amperometric detection; Regio-isomers