Emwas Abdul-Hamid M, Al-Talla Zeyad A, Yang Yang, Kharbatia Najeh M
NMR Core Lab, King Abdullah University of Science and Technology, Room 0149, Thuwal, 23955-6900, Kingdom of Saudi Arabia,
Methods Mol Biol. 2015;1277:91-112. doi: 10.1007/978-1-4939-2377-9_8.
Gas chromatography-mass spectrometry (GC-MS) has been widely used in metabonomics analyses of biofluid samples. Biofluids provide a wealth of information about the metabolism of the whole body and from multiple regions of the body that can be used to study general health status and organ function. Blood serum and blood plasma, for example, can provide a comprehensive picture of the whole body, while urine can be used to monitor the function of the kidneys, and cerebrospinal fluid (CSF) will provide information about the status of the brain and central nervous system (CNS). Different methods have been developed for the extraction of metabolites from biofluids, these ranging from solvent extracts, acids, heat denaturation, and filtration. These methods vary widely in terms of efficiency of protein removal and in the number of metabolites extracted. Consequently, for all biofluid-based metabonomics studies, it is vital to optimize and standardize all steps of sample preparation, including initial extraction of metabolites. In this chapter, recommendations are made of the optimum experimental conditions for biofluid samples for GC-MS, with a particular focus on blood serum and plasma samples.
气相色谱-质谱联用技术(GC-MS)已广泛应用于生物流体样本的代谢组学分析。生物流体提供了有关全身以及身体多个部位新陈代谢的丰富信息,可用于研究总体健康状况和器官功能。例如,血清和血浆能够提供全身的综合情况,尿液可用于监测肾脏功能,而脑脊液(CSF)则能提供有关大脑和中枢神经系统(CNS)状态的信息。已开发出不同方法用于从生物流体中提取代谢物,这些方法包括溶剂萃取、酸处理、热变性和过滤。这些方法在去除蛋白质的效率以及提取的代谢物数量方面差异很大。因此,对于所有基于生物流体的代谢组学研究而言,优化和标准化样品制备的所有步骤(包括代谢物的初始提取)至关重要。在本章中,针对用于GC-MS的生物流体样本,给出了最佳实验条件的建议,尤其着重于血清和血浆样本。