1. Supplemental Figures and Tables

2. WebMetabase: Batch Processor Settings
Supplementary Figure 1: Screenshots from WebMetabase Batch Processor showing the settings used for the evaluation of the metabolites for HT-MetId

3. Supplementary Figure 2: Metabolite identification with WebMetabase (WMB) The analysis of human and rat microsomal stability samples by LC/MS/MS with time points for 0, 5, 10, 15, 20 and 30 minutes show species differences in metabolization of Substrate 0 / parent (blue) into the two metabolites M17-14 / D 703 (red) and M / Norverapamil (orange) in human and rat. (left: human microsomal stability, right: rat microsomal stability; Substrate 0 / parent (blue); Metabolites: M17-14 / D703 RT= 1.37 (red) and M / Norverapamil RT=1.42 (orange)

4. Buspirone metabolism
Supplementary Figure 3: Microsomal metabolism of Buspirone in HT-MetId and Metd In microsomal incubations not only primary (1st generation) metabolites are observed, but at longer incubation time also multiple secondary (2nd generation metabolites are formed by metabolism of the primary metabolites. For the identification of the main site of metabolism these metabolites are not crucial.

5. Supplementary Table 1: Comparison of cycle times for Quan/Qual and separated workflow. Overall time needed for analysis and evaluation of 96 compounds using cassette of 6 compounds. Here we assumed that for MetID the same procedure with cassetted compounds is used as in our newly described workflow. Using individual runs would increase the analysis run time at least 6-fold.

6. Supplementary Table 2: Clearance values and signal intensity of the reference compounds used in the workflow validation Compounds were chosen to represent a large variety of chemical structures and a large range of microsomal clearance values. * Indicate compounds used also for the validation of the quantitative clearance determination on the QTOF
Clearance Human
[µl/min/mg]
Clearance Rat
[µl/min/mg]