低聚合度壳寡糖制备及其组分的色谱行为分析

Preparation and the chromatographic behavior analysis of low degree polymerization of Chitooligosaccharides components

摘要: 利用壳聚糖酶水解壳聚糖,再通过乙醇沉淀法制备低聚合度（Dp<6）的壳寡糖。利用ShodexNH2P-504E色谱柱,以乙腈-氨水为流动相梯度洗脱,建立了一种分离度好、灵敏度高的壳寡糖HPLC分析方法;利用UPLC-MS定性鉴定壳寡糖中N-乙酰化的组分。研究还探讨了壳寡糖组分的色谱行为,发现壳寡糖的N-乙酰化是导致壳寡糖不按照聚合度大小进行色谱分离的主要原因,为壳寡糖质量分析、组分分离提供了依据。

Abstract: Low- degree polymerization chitooligosaccharides ( COS) were prepared with the chitosan hyrolized by chitosan enzymatic utilizing the ethanol precipitation method.A HPLC analysis method was also presented for COS with remarkable separation and high sensitivity based on Shodex NH2P-50 4E Column and acetonitrile- ammonia mobile phase as the gradient elution.The N-acetylated constituent in COS could be identified using UPLC-MS.The chromatographic behavior of COS was also studied. Results illustrated that the N- acetylated constituent in COS was the main reason for that chromatographic separation of chitosan is not in accordance with its polymerization degree, which was very important for analyzing and separating the constituents of chitosan.