UHPLC system with high-resolution mass spectrometer (UHPLC-HRMS/MS)

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The research groups of the Institute of Pharmaceutical Biology investigate the biosynthesis, evolution and biological functions of plant specialized metabolites as well as methods for biotechnological production and qualitative and quantitative detection of these substances in food, feed and plant-derived pharmaceuticals. The analysis of these substances and their natural mixtures is a core element of all research projects. Analytical challenges are in particular (1) the small amounts of the analytes compared to typical primary metabolites (e.g. complex mixtures of differently substituted compounds, main and by-products of enzyme reactions, traces of toxic alkaloids), (2) the complex matrices (e.g. plant extracts), against which a sensitive detection of the substances has to be performed, (3) the instability of many analytes (e.g. polyprenylated acylphloroglucinols), (4) the detection of new or poorly characterized structures (e.g. new active compounds from St. John's wort or fiber hemp, biosynthetic intermediates and by-products), (5) the difficulty of identifying differences between the complex metabolite profiles of different samples (e.g. wild-type vs. mutant) and assigning structural information. Here, we apply for a UHPLC-HRMS/MS system that can help overcome these challenges. Ultra-High-Performance Liquid Chromatography (UHPLC) allows high chromatographic separation efficiency and resolution as an important prerequisite for sensitive detection. High-resolution mass spectrometry (HRMS) allows the determination of precise molecular masses and, in the case of the requested HR tandem mass spectrometry (HRMS/MS), also the precision mass determination of fragment ions. Using the precise masses, molecular and fragment ion molecular formulae can be determined and structural predictions can be made. This allows structural clues to be assigned to the signals obtained from the analytes. For known substances, identification may be possible. The requested instrument is equipped with metabolomics software that can be used to compare metabolite profiles, i.e. features from HRMS(/MS) assigned to a specific retention time, between multiple samples. The instrument is suitable for measurements of samples with complex matrices by providing a wide dynamic range. Thus it allows sensitive detection and quantification up to trace analysis of small molecules even in complex matrices. It replaces a technically obsolete instrument of a different type, which had been in operation for more than 15 years, and expands the existing work possibilities at the institute due to its high sensitivity, mass precision and chromatographic as well as mass spectrometric resolution coupled with its suitability for metabolomics approaches.