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

药物生物学研究所的研究小组研究植物专门代谢物的生物合成、进化和生物功能，以及食品、饲料和植物衍生药物中这些物质的生物技术生产和定性和定量检测方法。对这些物质及其天然混合物的分析是所有研究项目的核心要素。分析方面的挑战尤其在于：(1)与典型的初级代谢物（如不同取代化合物的复杂混合物，酶反应的主要和副产物，痕量的有毒生物碱）相比，分析物的数量很少，(2)复杂的基质（如植物提取物），必须对这些物质进行敏感的检测，(3)许多分析物的不稳定性（如聚戊烯酰化酰基间苯三酚），(4)检测新的或特征不明显的结构（例如，来自圣约翰草或大麻纤维的新的活性化合物，生物合成中间体和副产品），(5)识别不同样品（例如野生型与突变型）的复杂代谢物谱之间的差异和分配结构信息的困难。在这里，我们申请一个UHPLC-HRMS/MS系统，可以帮助克服这些挑战。超高效液相色谱（UHPLC）是实现高分离效率和高分辨率检测的重要前提。高分辨率质谱（HRMS）可以精确测定分子质量，在要求的HR串联质谱（HRMS/MS）的情况下，也可以精确测定片段离子的质量。利用精确的质量，可以确定分子和碎片离子分子式并进行结构预测。这允许结构线索被分配到从分析物获得的信号。对于已知物质，鉴定是可能的。所要求的仪器配备代谢组学软件，可用于比较多个样品之间的代谢物谱，即分配到特定保留时间的HRMS（/MS）特征。该仪器具有较宽的动态范围，适用于复杂基质样品的测量。因此，它允许敏感的检测和定量到痕量分析的小分子，甚至在复杂的矩阵。它取代了一种技术上已经过时的不同类型的仪器，该仪器已经运行了超过15年，并扩大了研究所现有的工作可能性，因为它具有高灵敏度、质量精度、色谱和质谱分辨率，以及它对代谢组学方法的适用性。

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.