Mass spectrometry method with C=C location specificity for qualitative and quantitative lipid analysis

 描述（由申请人提供）：在本项目中，我们的目标是开发一种质谱（MS）方法，该方法能够确定脂质中的碳-碳双键（C=C）位置，并将其应用于定性和定量脂质组学分析。该项目建立在将光化学反应Paternel-Büchi（PB）反应与电喷雾电离和串联质谱（ESI-MS/MS）耦合的创新基础上。通过碰撞诱导解离（CID），脂质的PB产物在原始C=C位置断裂，从而允许脂质C=C位置异构体的混合物的C=C位置测定和脂质定量。在该R 01项目中，建议开展研究活动，以开发和验证完全集成的PB-MS/MS方法，其目标如下：（1）开发和优化用于Paterness-Büchi（PB）反应与ESI-MS/MS的在线耦合的反应和电离条件，（2）开发用于不同类别的不饱和脂质的结构测定和定量的MS/MS方法，（3）PB-MS/MS方法在脂质组学分析中的应用与验证。普渡大学的Xia研究小组在分析方法，MS仪器和数据分析工具的开发方面具有专业知识。该项目的预期成果是为脂质组学提供一个具有C=C特异性的强大且广泛适用的MS平台。将产生一组关于大鼠/小鼠组织样品中内源性不饱和脂质的新知识，包括其C=C位置信息和C=C位置异构体的组成。这种新的脂质分析能力可以潜在地推进生物科学中许多领域的研究，包括但不限于脂质分子生物学、功能脂质组学、代谢组学和生物标志物发现。

 DESCRIPTION (provided by applicant): In this project, we aim to develop a mass spectrometry (MS) method which has the capability of determining carbon-carbon double bond (C=C) locations in lipids and apply it to qualitative and quantitative lipidomic analysis. This project builds upon an innovation of coupling a photochemical reaction, the Paternò-Büchi (PB) reaction, with electrospray ionization and tandem mass spectrometry (ESI-MS/MS). Through collision-induced dissociation (CID), the PB products of lipids fragment at the original C=C positions, allowing both C=C location determination and lipid quantitation for mixtures of lipid C=C position isomers. In this R01 project, research activities are proposed to develop and validate a fully integrated PB-MS/MS approach, with the following aims: (1) development and optimization of reaction and ionization conditions for on-line coupling of Paternò-Büchi (PB) reaction with ESI-MS/MS, (2) development of MS/MS methods for structural determination and quantitation of unsaturated lipids of different classes, and (3) implementation and validation of PB-MS/MS method for lipidomic analysis. The Xia research group at Purdue University has the expertise in the development of analytical methods, MS instrumentation, and data analysis tools. The expected outcome from the project is to provide a robust and widely applicable MS platform with C=C specificity for lipidomics. A set of new knowledge will be generated for unsaturated lipids endogenous in rats/mice tissue samples including their C=C position information and composition of C=C position isomers. This new lipid analysis capabilities can potentially advance research in many fields in biological sciences, including but not limited to lipid molecular biology, functional lipidomics, metabolomics, and biomarker discovery.