点击化学功能化POSS杂化整体柱电色谱新技术及抗生素肽残留分析

Food safety of peptide antibiotics (PTs) is the focus issue of concern to the international community. The residue analysis of PTs faces with the dual challenges of selectivity and stability, since they are kinds of polar and alkaline peptides with complex composition and structural similarity. It still lacks of an effective physics-chemical analysis method so far. In this project, a novel grafting approach based on the thiol-ene click reaction and organic-inorganic hybrid technique, will be introduced in the preparation and functionalization of POSS hybrid capillary monolithic stationary phase, according to the unique chemical structure and properties of PTs. The development of functionalized monolithic capillary electrochromatography hyphenated technique paves a new avenue for efficient, selective and sensitive analysis of PTs. The research strategy includes three aspects. For constructing of novel monolithic stationary phase of superior characteristics (hypercrosslinked, good permeability, stability and high column efficiency), optimization of the pore structure and surface area of the column will be performed by investigating the modes of initiated reaction, synthesis conditions of hybrid, and rules of click reaction. Owing to the regioselectivity and speedy of light-intiated click reaction, high efficient functionalization method of polar zwitterionic /mixed mode monolithic column can be developed, and used to improve the selectivity, separation efficiency and biocompatibility of stationary phase. Combining with the hyphenated technique of CEC with sensitive detection, high efficient and environmental friendly analysis of trace PTs residue can be achieved, and the mechanism of interaction and synergic separation between PTs and the stationary phase will be discussed, which provides an effective method for food safety monitoring and food safety assessment.

抗生素肽（PTs）的食品安全性是国际社会广泛关注的焦点问题。作为组成复杂、结构近似的极性碱性肽类，其残留分析面临选择性和稳定性的双重挑战，至今缺乏有效理化方法。课题针对PTs化学结构特性，基于巯基-烯点击化学功能化和有机无机杂化技术，探索功能化POSS杂化整体柱的制备新路径，发展高效、稳定的功能化杂化整体柱及毛细管电色谱（CEC）联用分析新技术。研究策略关键在于：通过点击反应聚合引发方式、杂化条件和规律研究，优化整体柱孔结构，构建高度交联、通透、稳定和柱效优越的新型整体固定相；利用光点击反应定域化和快速优势，发展两性/亲水混合模式整体柱的高效功能化方法，提高固定相的选择性、分离效能与生物相容性；阐明PTs与新型固定相的相互作用与协同分离机制，结合CEC-高灵敏检测联用技术，解决PTs分离难题，实现痕量PTs残留的高效、快速和环境友好的分析，为食品安全监控和安全性评价提供有力手段。

项目针对抗生素肽残留分析缺乏有效理化方法的瓶颈问题，重点创新研发杂化整体柱功能化制备路径和整体微萃取-毛细管电色谱（CEC）联用分析技术，实现了痕量抗生素肽的快速灵敏和环境友好分析。主要成果如下：.（1）基于巯基-烯点击化学、自由基聚合反应与有机无机杂化技术，以纳米级POSS、离子液体（ILs）以及两性或亲水性基团作为功能化单体，提出了功能化杂化整体柱的热引发、光引发与离子液体介质反应等多种高效的“one-pot”制备新路径和新技术，研制了系列高效、稳定和重现性好的功能化杂化整体分离柱与微萃取柱12种，包括C8-POSS杂化固定相、混合或亲水型离子液体功能化杂化柱、离子液体功能化POSS杂化柱以及磷酸基或磷酸胆碱功能化POSS杂化柱等不同相互作用类型。.开展了扫描电镜、红外光谱、氮吸附法、能谱分析、拉曼光谱和CEC表征，优化确定了整体柱功能化制备过程中影响电渗流、渗透性、柱效能的决定性因素，阐明了系列杂化整体柱对不同种类分析物的分离模式与机制。成果为杂化整体材料的高效功能化提供了绿色新途径，尤其是光化学引发或离子液体介质反应制备方法可以使制备时间缩短为几分钟，极大改善了传统整体柱制备周期或能耗方面的不足。.（2）构建了整体微萃取 (PMME) -CEC/CLC联用系统及在线/离线富集新技术，利用所研制的杂化整体微萃取柱，发展了抗生素肽的PMME-CEC/CLC- UV/LIF/AD联用分析新技术。优化影响分析物富集与分离效能的实验条件，阐明抗生素肽分离与富集机制，构建了功能化富集与选择性分离-高灵敏检测的联用新模式，并开展动物性样品分析与标准方法对照研究，实现了痕量抗生素肽残留的高灵敏、选择性分离分析。成果为抗生素肽的残留分析和食品安全监测提供了科学依据和环保经济的工具。.相关成果在Anal. Chem.、Electrophoresis、Microchim. Acta、J. Sep. Sci.、Sens. Actuat. B: Chem.、分析化学、色谱、有机化学等国内外重要专业期刊发表论文11篇, 其中SCI收录9篇，申请并获得国家发明专利授权 3项。参加国内外重要学术会议交流并做会议报告6次，培养博硕士研究生8名。项目已实现了预期目标。