Vorkas Panagiotis A, Abellona U M R, Li Jia V
Section of Biomolecular Medicine, Division of Computational and Systems Medicine, Department of Surgery and Cancer, Faculty of Medicine, Imperial College London, London, UK.
Centre for Digestive and Gut Health, Institute of Global Health Innovation, Imperial College London, London, UK.
Methods Mol Biol. 2018;1738:239-260. doi: 10.1007/978-1-4939-7643-0_17.
The use of tissue as a matrix to elucidate disease pathology or explore intervention comes with several advantages. It allows investigation of the target alteration directly at the focal location and facilitates the detection of molecules that could become elusive after secretion into biofluids. However, tissue metabolomics/metabonomics comes with challenges not encountered in biofluid analyses. Furthermore, tissue heterogeneity does not allow for tissue aliquoting. Here we describe a multiplatform, multi-method workflow which enables metabolic profiling analysis of tissue samples, while it can deliver enhanced metabolome coverage. After applying a dual consecutive extraction (organic followed by aqueous), tissue extracts are analyzed by reversed-phase (RP-) and hydrophilic interaction liquid chromatography (HILIC-) ultra-performance liquid chromatography coupled to mass spectrometry (UPLC-MS) and nuclear magnetic resonance (NMR) spectroscopy. This pipeline incorporates the required quality control features, enhances versatility, allows provisional aliquoting of tissue extracts for future guided analyses, expands the range of metabolites robustly detected, and supports data integration. It has been successfully employed for the analysis of a wide range of tissue types.
使用组织作为基质来阐明疾病病理学或探索干预措施具有多个优点。它允许在病灶部位直接研究目标改变，并有助于检测那些分泌到生物流体后可能难以捉摸的分子。然而，组织代谢组学/代谢物组学面临着生物流体分析中未遇到的挑战。此外，组织异质性不允许对组织进行等分。在此，我们描述了一种多平台、多方法的工作流程，该流程能够对组织样本进行代谢谱分析，同时可提供增强的代谢组覆盖范围。在进行连续两次提取（先有机提取，后水提取）后，通过反相（RP-）和亲水相互作用液相色谱（HILIC-）超高效液相色谱与质谱（UPLC-MS）以及核磁共振（NMR）光谱联用对组织提取物进行分析。该流程纳入了所需的质量控制功能，增强了通用性，允许对组织提取物进行临时等分以备将来的定向分析，扩大了能可靠检测的代谢物范围，并支持数据整合。它已成功用于分析多种组织类型。
