While global metabolic profiling (untargeted metabolomics) has been the center of much interest and research activity in the past few decades, more recently targeted metabolomics approaches have begun to gain ground. These analyses are, to an extent, more hypothesis-driven, as they focus on a set of pre-defined metabolites and aim towards their determination, often to the point of absolute quantification. The continuous development of the technological platforms used in these studies facilitates the analysis of large numbers of well-characterized metabolites present in complex matrices. The present review describes recent developments in the hyphenated chromatographic methods most often applied in targeted metabolomic/lipidomic studies (LC-MS/MS, CE-MS/MS, and GC-MS/MS), highlighting applications in the life and food/plant sciences. The review also underlines practical challenges-limitations that appear in such approaches.

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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.