Abstract:
Chitosan and its derivatives are interesting biopolymers for different field of analytical chemistry, especially in separation techniques. The present study was aimed at testing chitosan water soluble derivatives as dynamic coating agents for application to capillary electrophoresis. In particular, chitosan was modified following three different chemical reactions (nucleophilic substitution, reductive amination, and condensation) to introduce differences in charge and steric hindrance, and to assess the effect of these physico-chemical properties in capillary electrophoresis. The effects were tested on the capillary electrophoretic separation of the glycoforms of human transferrin, an important iron-transporting serum protein, one of which, namely disialo-transferrin (CDT), is a biomarker of alcohol abuse. Chitosan derivatives were characterized by using NMR and 1H NMR, HP-SEC-TDA, DLS, and rheology. The use of these compounds as dynamic coatings in the electrolyte running buffer in capillary electrophoresis was tested assessing the peak resolution of the main glycoforms of human transferrin and particularly of disialo-transferrin. The results showed distinct changes of the peak resolution produced by the different derivatives. The best results in terms of peak resolution were achieved using polyethylene glycol (PEG)-modified chitosan, which, in comparison to a reference analytical approach, provided an almost baseline resolution of disialo-transferrin from the adjacent peaks.
摘要:
壳聚糖及其衍生物是分析化学不同领域的有趣生物聚合物,特别是在分离技术方面。本研究旨在测试壳聚糖水溶性衍生物作为动态涂层剂在毛细管电泳中的应用。特别是,壳聚糖经过三种不同的化学反应(亲核取代,还原胺化,和冷凝)引入电荷和空间位阻的差异,并评估这些理化性质在毛细管电泳中的作用。测试了对人转铁蛋白糖型毛细管电泳分离的影响,一种重要的铁运输血清蛋白,其中之一,即双唾液酸转铁蛋白(CDT),是酒精滥用的生物标志物。用NMR和1HNMR对壳聚糖衍生物进行了表征,HP-SEC-TDA,DLS,和流变学。测试了这些化合物在毛细管电泳中的电解质运行缓冲液中作为动态涂层的用途,以评估人转铁蛋白,特别是二唾液酸转铁蛋白的主要糖型的峰分辨率。结果表明,不同衍生物产生的峰分辨率有明显变化。使用聚乙二醇(PEG)修饰的壳聚糖,在峰分辨率方面取得了最好的结果,which,与参考分析方法相比,提供了来自相邻峰的双唾液酸转铁蛋白的几乎基线分辨率。