在线联用银纳米颗粒固定化整体柱选择性富集与精准分析反式脂肪酸技术研究

Focused on the problem of the determination of trans fatty acid (TFA), we innovatively fabricate a novel silver nanoparticles (AgNPs) immobilized monolithic column for highly selective enrichment and accurate analysis of TFA, and present a sensitive online analysis method for TFA by utilizing the selective AgNPs monolithic column as stationary phase for both solid phase microextraction (SPME) and analytical chromatography column. The selective AgNPs monolithic column is fabricated by immobilized AgNPs on the active sites (e.g. amino, carboxyl and thiol groups) on the suface of the basic monolithic column which is prepared by rapid in-situ urea-formaldehyde polycondensation. The preparation feeds, preparation conditions and AgNPs immobiled conditions will be investigated respectively. A series of performance and characterization of the prepared monolithic column will be carried out. The structure of online analysis system for the determination will be designed by online coupling AgNPs-SPME with AgNPs-HPLC. The operating parameter of the system will be optimized, and the selectivity mechanisms of the AgNPs monolithic column will also be discussed. Taking the advantages of the specific interaction between AgNPs and unsaturated fatty acid, the online extraction of TFA and the selective and sensitive determination of TFA could be integrated into one step. A novel technology for online sensitive analysis of trace TFA in food will be developed to provide experimental data and scientific basis for people's health and risk assessment of food safety.

针对反式脂肪酸(TFA)的检测难题，本项目基于原位一步脱水快速缩聚的整体柱制备技术，创新性地提出了制备可选择性富集与分离TFA的银纳米颗粒(AgNPs)固定化整体柱，并结合基于AgNPs整体柱的固相微萃取-液相色谱(AgNPs-SPME-AgNPs-HPLC)在线联用系统，实现TFA精准分析。首先制备表面带活性位点的基质整体柱，通过AgNPs与活性位点相互作用实现整体柱表面AgNPs固定化，研究基质整体柱制备条件，优化AgNPs固定化条件，并对AgNPs整体柱进行性能测试。研究构建AgNPs-SPME-AgNPs-HPLC在线联用系统，利用AgNPs与不饱和脂肪酸之间的特殊相互作用，实现复杂样品中微量TFA选择性分步富集与精准分析，探讨TFA选择性富集与分离机理；优化系统运行条件，建立食品中微量TFA的高分离度高灵敏度分析新技术，为食品安全风险评估、保障人民身体健康提供实验数据和科学依据。

本项目基于酸催化原位一步脱水快速缩聚的整体柱制备技术，制备了透明质酸钠功能化脲醛树脂整体柱、银纳米颗粒表面涂覆整体柱、原位还原法制备的银纳米颗粒功能化整体柱等几种类型整体柱；并对制备的整体柱进行了全面的材料测试与表征，考察了包括反式脂肪酸在内的几类化合物的管内固相微萃取与微柱液相色谱分离行为与机理。本项目还利用室温席夫碱反应，开展了几种亚胺型共价有机框架材料的制备，通过对基质共价有机聚合物进行磺酸基功能化和银离子功能化，拓展了其在反式脂肪酸等化合物的富集分析应用。本项目进一步创制了以六通阀和十通阀为基础的在线管内固相微萃取-高效液相色谱（in-tube SPME-HPLC）联用系统，实现了该系统与二极管阵列和紫外等检测器的联用，并以三磷酸腺苷及其磷酸化代谢产物和唑来膦酸、利塞膦酸等二磷酸盐类药物等物质为研究对象，验证上述联用系统的稳定性和可靠性。以本项目开发的银纳米颗粒表面涂覆整体柱、原位还原法制备的银纳米颗粒功能化整体柱、以及银离子功能化共价有机聚合物作为固相微萃取介质，依靠银纳米颗粒与不饱和双键的银离子型相互作用以及银离子与不饱和双键的电荷转移相互作用，实现反式脂肪酸等检测对象的在线选择性富集，并结合in-tube SPME-HPLC联用系统，发展了几种不同类型食品中的微量反式脂肪酸在线富集分析新技术。此外，本项目还以原位还原法制备的银纳米颗粒功能化整体柱为微柱液相色谱分析柱，利用银纳米颗粒与不饱和π键系统的银离子型相互作用，实现了包括反式脂肪酸在内的几种不饱和化合物的高效识别分析，进一步发掘了银纳米颗粒功能化整体柱的分离应用潜力。本项目共发表SCI论文10篇，还有2篇文章正在整理，共申请发明专利12项，授权发明专利7项，授权实用新型专利3项，发表会议论文3篇，并作国内会议报告2场，国际会议报告1场，培养研究生12名。各项指标均超出预期目标，圆满完成了此项课题。