The discovery of extracellular vesicles (EVs), including exosomes, microvesicles, and apoptotic bodies, has opened a new frontier in the study of signal transduction and understanding cell-to-cell communication. EVs play a key role regulating various biological processes such as tissue regeneration, blood coagulation, immunomodulation or carcinogenesis. Therefore, they gain much attention as potentially therapeutic and non-invasive diagnostic targets. In this aspect, a comprehensive characterization of the EVs molecular content and understanding EVs functions are especially relevant. However, routine isolation of a usually small amount of high purity EVs is challenging, and analysis requires highly sensitive advanced analytical solutions. The utility of EVs in clinical settings is still limited due to the lack of standardization in existing protocols. This review focuses on the implication of the state-of-the-art mass-spectrometry based untargeted metabolomics approach for the molecular profiling of EVs, with the emphasis on the analytical methodologies, highlighting potential challenges and limitations.

细胞外囊泡 (EV) 的发现，包括外泌体、微囊泡和凋亡小体，为信号转导和理解细胞间通讯的研究开辟了新领域。EVs 在调节组织再生、血液凝固、免疫调节或致癌作用等各种生物过程中发挥着关键作用。因此，它们作为潜在的治疗和非侵入性诊断目标而备受关注。在这方面，EVs 分子含量的综合表征和理解 EVs 功能尤其重要。然而，对通常少量的高纯度 EV 进行常规分离具有挑战性，并且分析需要高度灵敏的高级分析解决方案。由于现有协议缺乏标准化，EV 在临床环境中的实用性仍然有限。本综述侧重于基于最先进质谱的非靶向代谢组学方法对 EV 进行分子分析的意义，重点是分析方法，强调潜在的挑战和局限性。