Metabolic profiling performed using untargeted metabolomics of different, complex biological samples aims to apply agnostic/holistic, hypothesis-free, analysis of the small molecules that are present in the analyzed sample. This approach has been the center of major investments and dedicated efforts from the research community for many years. However, limitations and challenges remain, particularly with regard to the validation and the quality of the obtained results. This has led to increasing community engagement, with the formation of think tanks, the establishment of working groups, and the many seminars on quality control (QC) in metabolomics. Here we describe a quality control (QC) protocol used to monitor LC-MS-based metabolomics analysis. A key target is the monitoring of analytical precision. This methodology is described for the analysis of urine but can be applied to different biological matrices, such as various biofluids, cell, and tissue extracts.
Keywords: Biological samples; LC-MS; Quality control; Untargeted metabolomics; Validation.
© 2025. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.