Liquid chromatography electrospray ionisation triple quadrupole mass spectrometer (LC-ESI-QQQ)

有机化合物的定性和定量分析在营养科学领域至关重要，可以表征（微）营养素的代谢及其在信号传导途径中的影响，并确定营养状况和饮食相关疾病风险的生物标志物。在此背景下，本文应用的液相色谱耦合电喷雾电离三重四极杆质谱仪（LC-ESI-QQQ-MS）将主要用于蛋白质和多肽的定量以及生物样品（血液和组织样品，细胞培养等）中翻译后修饰（PTMs）的表征，具体考虑蛋白质氧化和蛋白质磷酸化。与经典的基于抗体的方法（如ELISA或Western blot）相比，基于lc - esi - qqq - ms的方法提供了区分、表征（PTM位点）和量化蛋白质的各种PTM的可能性。此外，它们允许建立高通量方法，同时减少样本量。除了对PTMs进行定性和定量分析外，LC-ESI-QQQ-MS还可以检测可能产生的蛋白质形态，例如，由可变剪接或可变启动子产生的蛋白质形态。不同的蛋白质形态在结构、定位和功能上有很大的不同，对新陈代谢和体内平衡的影响也不同。因此，它们的系统表征可以更好地评估代谢状态，有助于更好地理解（营养相关）慢性疾病的发病机制和病理生理，从而实施创新和个性化的预防和治疗措施。除了营养生理学主席（Henze教授），基于lc - esi - qqq - ms的有机分子分析也与人类营养学（Stangl教授）、二级植物化合物生物功能（Wätjen教授）和动物营养学（Zeyner教授）教授的创新研究方法密切相关。因此，拟议的LC-ESI-QQQ-MS不仅将用于蛋白质和多肽的分析，而且还将用于微量营养素（例如维生素D）和次生植物化合物的定量和表征——尤其是评估它们对内源性蛋白质的影响。农业和营养科学研究所和大学现有设备的能力已经完全耗尽，无法满足日益增加的需求。此外，现有设备的灵敏度和测量精度仅部分达到现行标准。因此，为了保持高质量的研究，在第三方资助领域保持竞争力，并保持合作的吸引力，所要求的LC-ESI-QQQ-MS是强制性的。

The qualitative and quantitative analysis of organic compounds is of central importance in the field of nutritional science to characterize the metabolism of (micro-)nutrients and their implications in signaling pathways, and to determine biomarkers for nutritional status and risk of diet-related diseases. In this context, the liquid chromatography-coupled electrospray ionization triple quadrupole mass spectrometer (LC-ESI-QQQ-MS) applied for here will be used primarily for the quantification of proteins and peptides as well as the characterization of post-translational modifications (PTMs) in biological samples (blood and tissue samples, cell culture etc.), with specific consideration of protein oxidation and protein phosphorylation. In contrast to classical antibody-based methods such as ELISA or Western blot, LC-ESI-QQQ-MS-based methods offer the possibility to differentiate, characterize (PTM site) and quantify various PTMs of a protein. Furthermore, they allow the establishment of high-throughput approaches while reducing the sample volume. In addition to the qualitative and quantitative analysis of PTMs, LC-ESI-QQQ-MS also enables the detection of proteoforms that can arise, for example, from alternative splicing or variable promoters. The different proteoforms can vary considerably in structure, localization, and function and can affect metabolism and homeostasis differently. Their systematic characterization therefore allows a better assessment of the metabolic status and contributes to a better understanding of the pathogenesis and pathophysiology of (nutrition-related) chronic diseases and thus to the implementation of innovative and individualized prevention and therapy measures.In addition to the Chair of Nutritional Physiology (Prof. Dr. Henze), LC-ESI-QQQ-MS-based analysis of organic molecules is also highly relevant for the professorships of Human Nutrition (Prof. Dr. Stangl), Biofunctionality of Secondary Plant Compounds (Prof. Dr. Wätjen) and Animal Nutrition (Prof. Dr. Zeyner) to work on innovative research approaches. The proposed LC-ESI-QQQ-MS will therefore be used not only for the analysis of proteins and peptides but also for the quantification and characterization of micronutrients (e.g. vitamin D) and secondary plant compounds - not least to assess their influence on endogenous proteins. The capacity of the equipment available at the Institute of Agricultural and Nutritional Science and the University, respectively, is already completely exhausted and will not meet the increasing demand. In addition, the sensitivity and measurement accuracy of the available equipment only partially meet current standards. The requested LC-ESI-QQQ-MS is therefore mandatory in order to perpetuate high-quality research, to remain competitive in the field of third-party funding and to maintain the attractiveness for collaborations.