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

克里克是一个现代化的医学研究所，拥有120多个研究实验室。克里克成为世界领先的研究机构之一的核心是使用最先进的仪器提供尖端的科学。为了实现这一目标，克里克建立了集中的科学技术平台（stp）来容纳这些仪器，并由具有特定专业知识的工作人员提供支持。这种模式允许操作效率和机构范围内一致的最高质量标准。在过去的十年中，代谢组学——测量给定生物系统中所有小分子（或代谢物）——已经成为基础细胞生物学研究的核心和必要技术。事实上，克里克代谢组学STP目前与约70个克里克研究实验室合作，对其中许多实验室来说，代谢组学对业务至关重要（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“广谱”发现分析（测量尽可能广泛的代谢物范围）到复杂的目标分析（例如代谢途径通量测量）。代谢物本身的化学和物理性质差别很大，从小的极性代谢物到大的非极性脂质。因此，代谢组学依赖于复杂的质谱仪器，以便尽可能多地覆盖广泛的代谢物。为了实现这一目标，代谢组学STP目前拥有三个液相色谱-质谱（LC-MS）仪器。这些Thermo“Orbitrap”仪器在代谢组学研究领域是公认的金标准。orbitrap为这项工作提供了必要的速度和质量分辨率，允许在任何给定的运行中检测和定量数千种代谢物，更重要的是，可以更好地识别“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，其中一半采用LC-MS分析。自2013年以来，我们的Thermo Q-Exactive Orbitrap仪器已经运行了36,000次，仅在过去两年中就有15,000次。这不仅突出表明这是一个主力工具，而且表明它的工作量在继续增加。然而，该仪器现在已处于使用寿命末期，考虑到仪器的工作量和随着时间的推移发生重大故障的风险增加，Q-Exactive的故障对代谢组学STP的必要功能来说是一个关键风险。此外，该系统缺乏新一代Orbitrap LC-MS仪器的分析能力。拟议的替代品Thermo Vanquish-Exploris 240将与目前代谢组学STP中其他仪器的功能相匹配。这样既可以保持数据质量标准的连续性，又无需对STP工作人员进行额外培训，因为他们在这一工具方面已经非常有经验。重要的是，exploris240的规格比Q-Exactive好得多。大大提高的质量分辨率和扫描速度对于我们专门研究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，并更深入地研究代谢和代谢途径活性。此外，Exploris 240改进了数据采集能力，包括“Acquire X”模式，该模式允许自动生成和更新离子碎片（“包含”和“排除”）列表的工作流程，允许不干预，更深入的特征分析，极大地提高了代谢物识别的效率和信心，提高了容量，为操作人员节省了宝贵的时间。

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.