基于稳定同位素标记技术的磷脂高选择性、高灵敏定性和定量分析新方法及其应用研究

In view of the technical bottlenecks of existing MS technologies on phospholipids analysis, this project is intended to integrate stable isotope labeling technique with electrospray ionization tandem mass spectrometry (ESI-MS/MS) and to establish a novel method based on stable isotope labeling technique for high selective and sensitive qualitative and quantitative analysis of phospholipids. Firstly, a series of stable isotope labeling reagents will be synthesized, which can be easily ionized with ESI-MS and can be used to react with the specific groups of phospholipids molecules efficiently and selectively; then rapid sample preparation methods for phospholipids are tried to be developed; meanwhile, shotgun lipidomic strategy will be used to establish a high selective and sensitive new method based on stable isotope labeling for global analysis of phospholipids; on this basis, the established platform will be applied for study of lipid nutrition. On one hand, analysis the changes of phospholipids and their metabolites content in rat plasma and liver tissues under variable fatty acid composition of dietary lipids, and therefore many characteristic phospholipids can be identified and molecular markers or metabolic rules can be uncovered; on the other hand, through preliminary study the molecular mechanisms of variable fatty acid composition ratio dietary on mediation of phospholipids metabolism so as to provide scientific basis for the reasonable intake of fatty acids. In conclusion, the research of this project will provide the new approach for qualitative and quantitative analysis of complex phospholipids compounds, and provides the theoretical basis and technological support for lipid nutrition research.

针对现有质谱技术在磷脂的分析检测中存在的技术瓶颈，本课题拟将稳定同位素标记技术与电喷雾串联质谱（ESI-MS/MS）相结合，开展基于稳定同位素标记技术的磷脂高选择性、高灵敏定性和定量分析新方法研究。合成一系列在ESI条件下易电离，能高效、且选择性的与磷脂分子中特定基团反应的稳定同位素标记试剂；开发针对磷脂类化合物的快速通用的样品预处理方法；采用 “鸟枪法”研究策略，建立基于稳定同位素标记技术的磷脂的高选择性、高灵敏的定性和定量分析新方法；在此基础上，将建立的分析平台应用于脂质营养研究中，分析不同脂肪酸构成比的膳食对大鼠血浆及肝脏组织中磷脂及其代谢路径上各组分含量的变化，找到相关的特征磷脂成分和标志分子或代谢规律，初步探讨不同脂肪酸构成比的膳食对磷脂脂质代谢调节作用的分子机制，为合理食用脂肪酸提供科学依据。本项目的研究将为磷脂等复杂样品的高效定性和定量分析提供新途径，为脂质营养研究提供理论依

针对现有质谱技术在磷脂的分析检测中存在的技术瓶颈，本课题将稳定同位素标记技术与电喷雾串联质谱（ESI-MS/MS）相结合，开展了基于稳定同位素标记技术的磷脂高选择性、高灵敏定性和定量分析新方法研究。开发了针对磷脂类化合物的快速通用的样品预处理方法；采用一系列在ESI条件下易电离，能高效、且选择性的与磷脂分子中特定基团反应的稳定同位素标记试剂，结合 “鸟枪法”研究策略，建立了基于稳定同位素标记技术的磷脂的高选择性、高灵敏的定性和定量分析新方法；在此基础上，将建立的分析平台应用于脂质营养研究中，分析不同脂肪酸构成比的膳食对大鼠血浆和组织中磷脂及其代谢路径上各组分含量的变化，找到相关的特征磷脂成分和标志分子或代谢规律，初步探讨不同脂肪酸构成比的膳食对磷脂脂质代谢调节作用的分子机制，为合理食用脂肪酸提供科学依据。本项目的研究将为磷脂等复杂样品的高效定性和定量分析提供新途径，为脂质营养研究提供理论依据。