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

• Subject Area: Medicine
• Term: Funded in 2022
• Project identifier: Deutsche Forschungsgemeinschaft (DFG) - Project number 516933750

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.

• DFG Programme: Major Research Instrumentation
• Major Instrumentation: Flüssigchromatographie-gekoppeltes Elektrosprayionisations Triple-Quadrupol-Massenspektrometer
• Instrumentation Group: 1700 Massenspektrometer
• Applicant Institution: Martin-Luther-Universität Halle-Wittenberg
• Leader: Professorin Dr. Andrea Henze