Wide-targeted Metabolomics in Dried Blood Spots

Dried blood spot (DBS) sampling offers a minimally invasive, logistically flexible alternative to venipuncture, and is suitable for remote metabolomics applications. This study presents a high-throughput workflow using volumetric absorptive microsampling (VAMS, 10 µL), miniaturized liquid–liquid extraction in well plates, and dual-column liquid chromatography–high-resolution mass spectrometry (LC–HRMS) for targeted metabolomics. Accurate quantification was enabled by fixed sample volume and yeast-derived fully ¹³C-labeled internal standards. The simplified extraction—shaking and filtration—minimized solvent use and enabled direct LC–HRMS injection. The method was validated using standard reference material (SRM) 1950 human plasma and assessed for DBS stability under room temperature and -80°C storage for 90 days.