High-resolution mass spectrometer with Liquid Chromatography (LC) and Capillary Electrophoresis (CE)

通过液-质联用(LC-MS)分离化合物的质谱学已经成为大规模代谢组学测量的关键方法。与直接进样质谱仪相比，LC-MS具有几个优点，因为复杂样品基质中的化合物被分离到不同的峰，从而减少了离子抑制并提高了灵敏度。这项提议旨在采购一台新的串联质谱仪，用于LC-MS和毛细管电泳质谱(CE-MS)小分子的测量，采用定向和非定向策略。新仪器将用于鉴定和定量信号分子(例如，肌醇磷脂和肌醇磷酸盐)、次生植物代谢物(例如，类黄酮类)和其他分子，重点是植物和细菌。我们将使用LC-MS建立一条管道，用于鉴定在两个生物系统(非靶标代谢组学)之间差异积累的代谢物，例如通过比较野生类型和突变体，或暴露在控制或应激条件下的生物。仪器将切换到CE-MS模式，用于测定肌醇-磷酸和其他极性、阴离子代谢物(NADPH、ATP等)。波恩大学的校园目前还不能使用CE-MS系统。质谱仪将安装并集成到波恩大学植物分子生理学和生物技术研究所(IMBIO)现有的脂质组学/代谢组学平台中。该平台已经运行了两台LC-MS仪器，用于甘油脂类的直接输注/鸟枪测量和低丰度代谢物的LC-MS分析。新的质谱仪将用于高质量精度和灵敏度的测量，这是鉴定和定量复杂提取物中大量代谢物所必需的。该文书将对已获得资金或正在评估的若干(联合)赠款申请至关重要。质谱仪将服务于校园内的团体，但也将开放给与国内和国际研究团体的合作。

Mass spectrometry with separation of compounds by e.g. liquid chromatography (LC-MS) has emerged as a key method for large scale metabolomic measurements. In contrast to direct infusion mass spectrometry, LC-MS offers several advantages because compounds from complex sample matrices are separated into different peaks, which reduces ion suppression and results in improved sensitivity. This proposal aims at the procurement of a new tandem mass spectrometer for LC-MS and capillary electrophoresis-mass spectrometry (CE-MS) measurements of small molecules, both employing targeted and non-targeted strategies. The new instrument will be used for the identification and quantification of signaling molecules (e.g., phosphoinositides and inositol-phosphates), secondary plant metabolites (e.g., flavonoids) and other molecules, with a focus on plants and bacteria. We will use the LC-MS to establish a pipeline for the identification of metabolites that differentially accumulate between two biological systems (non-targeted metabolomics), e.g. by comparing wild types and mutants, or organisms exposed to control or stress conditions. The instrument will be switched to CE-MS mode for the measurement of inositol-phosphates and other polar, anionic metabolites (NADPH, ATP, etc.). The CE-MS system is currently not available on campus at the University of Bonn. The mass spectrometer will be located and integrated into the existing Lipidomics/Metabolomics Platform at the Institute of Molecular Physiology and Biotechnology of Plants (IMBIO) of the University of Bonn. This platform already operates two LC-MS instruments, which are used at high capacity for direct infusion/shotgun measurements of glycerolipids, and LC-MS analysis of low abundant metabolites. The new mass spectrometer will be used for measurements with high mass accuracy and sensitivity, which is required for the identification and quantification of large numbers of metabolites in complex extracts. The instrument will be essential for several (joint) grant applications that have been funded or are under evaluation. The mass spectrometer will serve groups on campus, but it will also be open to collaborations with national and international research groups.