Liquid Chromatography-High Resolution Mass Spectrometer (LC-HRMS) for Discovery and Targeted Metabolomics/Lipidomics research

克里克研究所是一个拥有120多个研究实验室的现代医学研究所。实现克里克成为世界领先研究机构之一的雄心壮志的核心是使用最先进的仪器提供尖端科学。为了实现这一目标，克里克建立了中央科学技术平台(STP)，这些平台容纳了这种仪器，并得到了特定专业人员的支持。在过去的十年里，代谢组学--对给定生物系统中所有小分子(或代谢物)的测量--已成为基础细胞生物学研究中的一项核心和关键技术。事实上，克里克代谢组学STP目前与大约70个克里克研究实验室合作，对其中许多实验室来说，代谢组学是商业关键(特别是那些研究癌症代谢、癌症进展、感染和免疫、疾病发病机制和发育生物学的实验室)。代谢组学的范围可以从“广泛的”发现概况(测量尽可能广泛的代谢物)到复杂的目标分析(例如，代谢途径流量测量)。代谢物本身的化学和物理性质差异很大，从小的极性代谢物到大的非极性脂类。因此，代谢组学依赖于复杂的质谱学仪器，以尽可能多地涵盖这种广泛的代谢物。为了实现这一目标，代谢组学STP目前拥有三台液质联用(LC-MS)仪器。这些Thermo‘Orbitrap’仪器在该领域被公认为代谢组学研究的黄金标准。Orbitraps提供了这项工作所必需的速度和质量分辨率，允许在任何给定的运行中检测和量化数千种代谢物，而且--重要的是--更好地识别“未知”代谢物(很容易成为当今代谢组学的最大瓶颈)。STP每年运行约35,000个样品，其中一半由LC-MS分析。自2013年以来，在我们的Thermo Q-Exactive Orbitrap仪器上运行了36,000次，仅在过去两年就运行了15,000次。这不仅突显了这是一个主要的工具，而且它的工作量继续增加。然而，这个工具现在处于使用寿命的末期，考虑到工具的工作量和随着时间的推移重大故障风险的增加，Q-Exactive的故障对代谢组学STP发挥必要作用的能力来说是一个严重的风险。此外，该系统缺乏新一代Orbitrap LC-MS仪器的分析能力。拟议的替代仪器--Thermo Vanquish-Exploris 240--将与代谢组学STP中目前的其他仪器的能力相匹配。这将允许数据质量标准的连续性，并需要对STP工作人员进行微不足道的额外培训，因为他们已经在该仪器方面经验丰富。重要的是，Exploris 240的规格比Q-Exactive好得多。大大提高的质量分辨率和扫描速度将对我们专门从事的稳定同位素标记技术非常重要，使我们能够区分不同元素的同位素，并更深入地研究新陈代谢和代谢途径活动。此外，Exploris 240还改进了数据采集能力，包括“Acquire X”-一种允许自动生成和更新离子碎裂(“包含”和“排除”)列表的工作流程的模式，允许不插手、更深入的特征分析，极大地提高了代谢物识别的效率和置信度，提高了能力，为操作员腾出了宝贵的时间。

The Crick is a modern medical research institute that houses 120+ research laboratories. Central to meeting The Crick's aspiration of becoming one of the world's leading research institutes is the delivery of cutting-edge science using state-of-the-art instrumentation. To achieve this, The Crick has established centralised Scientific Technology Platforms (STPs) that house this instrumentation and are supported by staff of specific expertise. This model allows operational efficiency and institute-wide consistent standards of the highest quality.Over the last decade, metabolomics - the measurement of all small molecules (or metabolites) in a given biosystem - has emerged as a core and essential technology in fundamental cell biology research. Indeed, The Crick Metabolomics STP currently works with ~70 Crick research laboratories and for many of these, metabolomics is business critical (notably, those studying cancer metabolism, cancer progression, infection and immunity, disease pathogenesis, and developmental biology). Metabolomics can range from 'broad-stroke' discovery profiling (where as wide a range of metabolites as possible is measured) to intricate targeted analyses (e.g. metabolic pathway flux measurements). The metabolites themselves vary greatly in their chemical and physical properties, from small polar metabolites to large non-polar lipids. Metabolomics relies, therefore, on sophisticated mass spectrometry instrumentation in order to cover as much of this broad range of metabolites as possible. To achieve this, The Metabolomics STP currently houses three liquid chromatography-mass spectrometry (LC-MS) instruments. These Thermo 'Orbitrap' instruments are well-established in the field as being the gold standard for metabolomics studies. The Orbitraps afford the speed and mass resolution power essential for this work, allowing the detection and quantification of thousands of metabolites in any given run and - importantly - better identification of 'unknown' metabolites (easily the biggest bottleneck in metabolomics today).The STP runs ~35,000 samples per year, of which half are analysed by LC-MS. Since 2013, 36,000 have been run on our Thermo Q-Exactive Orbitrap instrument, with 15,000 in the last two years alone. This highlights not only that this is a workhorse instrument, but that its workload continues to increase.However, this instrument is now at end-of-life and, given both the workload of the instrument and the increased risk of major failure over time, breakdown of the Q-Exactive represents a critical risk to the ability of the Metabolomics STP to function as necessary. Moreover, this system lacks the analytical capabilities of the newer generation of Orbitrap LC-MS instruments. The proposed replacement - the Thermo Vanquish-Exploris 240 - will match the capabilities of the other instrumentation currently within the Metabolomics STP. This will both allow a continuity of data quality standards and require negligible additional training for the STP staff as they are already highly experienced in this instrument.Importantly, the Exploris 240 has considerably better specifications than the Q-Exactive. The much-improved mass resolution and scan speed will be important for the stable isotope labelling techniques in which we specialize, allowing us to differentiate between isotopes of different elements and study metabolism and metabolic pathway activity with greater depth. Moreover, the Exploris 240 has improved data acquisition capabilities, including 'Acquire X' - a mode that allows automated generation and updating of workflows for ion fragmentation ('inclusion' and 'exclusion') lists, allowing a hands-off, more in-depth feature analysis, hugely improving metabolite identification efficiency and confidence, improving capacity, and freeing up valuable time for the operator.