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

具有 C=C 位置特异性的质谱方法，用于定性和定量脂质分析

• 批准号: 9080382
• 负责人: Yu Xia
• 金额: $ 31.37万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-07-01 至 2021-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9080382
• 关键词: Benchmarking; Biochemical; Biological; Biological Process; Biological Sciences; Carbon; Cell membrane; Collaborations; Coupling; Data Analyses; Development; Diagnostic; Dissociation; Electrospray Ionization; Epigenetic Process; General Population; Goals; Homeostasis; Investigation; Ions; Isomerism; Knowledge; Lead; Lipids; Location; Mammalian Cell; Mass Spectrum Analysis; Medical; Methods; Modification; Molecular Biology; Molecular Structure; Mus; Outcome; Pathway interactions; Physiological; Positioning Attribute; Public Health; Rattus; Reaction; Reporting; Research; Research Activity; Sampling; Science; Series; Signal Transduction; Specificity; Spectrometry, Mass, Electrospray Ionization; Techniques; Tissue Sample; Universities; Unsaturated Fats; Validation; Veterinary Medicine; Work; analytical method; base; biomarker discovery; college; disease diagnosis; innovation; instrument; instrumentation; ionization; ionization technique; metabolomics; method development; pi bond; public health relevance; tandem mass spectrometry; tool

 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.

 描述(申请人提供)：在这个项目中，我们的目标是开发一种能够确定脂类中碳-碳双键(C=C)位置的质谱学(MS)方法，并将其应用于定性和定量脂肪组学分析。该项目建立在将光化学反应--帕特诺-比奇(PB)反应与电喷雾电离和串联质谱仪(ESI-MS/MS)相结合的创新基础上。通过碰撞诱导解离(CID)，脂类的PB产物在原始的C=C位置上碎裂，从而对脂类C=C位置异构体的混合物进行C=C位置测定和脂类定量。在这个R01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，目的如下：(1)开发和优化用于Paternó-Büchi(PB)反应与ESI-MS/MS在线耦合的反应和电离条件，(2)开发用于不同类别不饱和脂肪结构确定和定量的MS/MS方法，以及(3)实施和验证PB-MS/MS方法用于脂组分析。普渡大学的夏研究小组在分析方法、MS仪器和数据分析工具的开发方面拥有专业知识。该项目的预期结果是为脂质组学提供一个具有C=C特异性的强大和广泛适用的MS平台。大鼠组织中内源性不饱和脂类的C=C位置信息和C=C位置异构体的组成将产生一套新的知识。这种新的脂类分析能力可能会促进许多生物科学领域的研究，包括但不限于脂类分子生物学、功能脂类组学、代谢组学和生物标记物的发现。