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，以及生物技术生产和定性和定量检测食品、饲料和植物衍生药物中这些物质的方法。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内容。分析挑战尤其是(1)与典型的初级代谢物(例如，不同取代化合物的复杂混合物、酶反应的主要和副产物、有毒生物碱的微量)相比，分析物的量很少，(2)必须对物质进行灵敏检测的复杂基质(例如，植物提取物)，(3)许多分析物(例如，多戊烯基间苯三酚)不稳定，(4)检测到新的或特征不佳的结构(例如，来自圣约翰麦芽汁或纤维大麻的新的活性化合物，生物合成中间体和副产品)，(5)难以识别不同样品(例如野生型和突变型)的复杂代谢物图谱之间的差异，以及分配结构信息的困难。在这里，我们申请一种UHPLC-HRMS/MS系统，它可以帮助克服这些挑战。超高性能液相色谱(UHPLC)允许高的色谱分离效率和分辨率作为灵敏检测的重要前提。高分辨率质谱仪(HRMS)可以测定精确的分子质量，在要求的HR串联质谱仪(HRMS/MS)的情况下，还可以对碎片离子进行精确的质量测定。使用精确的质量，可以确定分子和碎片离子分子式，并可以进行结构预测。这允许将结构线索分配给从分析物获得的信号。对于已知的物质，鉴定是可能的。所要求的仪器配备了代谢组学软件，可用于比较多个样品之间的代谢物图谱，即分配给特定保留时间的HRMS(/MS)的特征。该仪器具有较宽的动态范围，适用于复杂矩阵样品的测量。因此，即使在复杂的基质中，它也可以对小分子进行灵敏的检测和定量，甚至可以进行痕量分析。它取代了已经运行了15年以上的另一种类型的技术上过时的仪器，并扩大了研究所现有工作的可能性，因为它具有高灵敏度、质量精确度和色谱学以及质谱学分辨率，再加上它适用于代谢组学方法。

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.