Fig. 1: DIAMetAlyzer - a pipeline for assay library generation and targeted analysis with statistical validation.

DDA data is used for candidate identification containing feature detection, adduct grouping and accurate mass search. Library construction uses fragment annotation via compositional fragmentation trees (SIRIUS) and decoy generation using a fragmentation tree re-rooting method (Passatutto) to create a target-decoy assay library. This library is used in a second step to analyse metabolomics DIA data by performing targeted extraction (OpenSWATH), scoring and statistical validation (PyProphet).