Global metabolic profiling (untargeted metabolomics) of different and complex biological matrices aims to implement an holistic, hypothesis-free analysis of (potentially) all the metabolites present in the analyzed sample. However, such an approach, although it has been the focus of great interest over the past few years, still faces many limitations and challenges, particularly with regard to the validation and the quality of the obtained results. The present protocol describes a quality control (QC) procedure for monitoring the precision of the analytical process involving untargeted metabolic phenotyping of urine and plasma/serum. The described/suggested methodology can be applied to different biological matrices, such as biological biofluids, cell, and tissue extracts.