Protocol for a low-volume, direct analysis urine preparation procedure for non-targeted GC-MS metabolomics.

We present a low-volume, direct analysis protocol for non-targeted gas chromatography-mass spectrometry (GC-MS) metabolomics, using 100 μL of urine. The steps include sample collection, stock solution preparation, metabolite extraction, two-step derivation with a drying phase, and analysis via two-dimensional GC time-of-flight MS (GCxGC-TOFMS). This protocol improves the efficiency and thoroughness of urinary metabolite analysis, contributing to advancements in metabolomics research, disease diagnosis, and biomarker discovery. For complete details on the use and execution of this protocol, please refer to Olivier et al.(1).