Institute of Medical Science, Tokyo Medical University, Tokyo, Japan.
Institute for Advanced Biosciences, Keio University, Tsuruoka, Yamagata, Japan.
Methods Mol Biol. 2023;2571:83-94. doi: 10.1007/978-1-0716-2699-3_8.
Capillary electrophoresis-mass spectrometry (CE-MS) is an ideal method for analyzing various metabolites in biological samples. CE-MS can simultaneously identify and quantify hundreds of charged metabolites using only two acquisition methods for positively and negatively charged metabolites. Furthermore, CE-MS is commonly used for analyzing biological samples to understand the pathology of diseases at the metabolic level and biofluid samples, such as blood and urine, to explore biomarkers. Here, we introduce a protocol that delineates the handling of clinical samples to ensure that the CE-MS analysis yields reproducible quantified data. We have focused on sample collection, storage, processing, and measurement. Although the implementation of rigorous standard operating protocols is preferred for enhancing the quality of the samples, various limitations in an actual clinical setting make it difficult to adhere to strict rules. Therefore, the effect of each process on the quantified metabolites needs to be evaluated to design a protocol with acceptable tolerances. Furthermore, quality controls and assessments to handle clinical samples are introduced.
毛细管电泳-质谱联用(CE-MS)是分析生物样品中各种代谢物的理想方法。CE-MS 仅使用两种正、负离子采集方法,即可同时鉴定和定量数百种带电荷的代谢物。此外,CE-MS 常用于分析生物样品,以了解疾病在代谢水平上的病理,以及血和尿等生物流体样本,以探索生物标志物。在这里,我们介绍了一个方案,阐述了临床样品的处理,以确保 CE-MS 分析产生可重复的定量数据。我们重点介绍了样品的采集、储存、处理和测量。虽然实施严格的标准操作方案有助于提高样品的质量,但在实际临床环境中存在各种限制,难以严格遵守规则。因此,需要评估每个过程对定量代谢物的影响,以设计具有可接受公差的方案。此外,还介绍了处理临床样品的质量控制和评估。