Additional file 1 of Solanidine is a sensitive and specific dietary biomarker for CYP2D6 activity

Additional file 1. Figure S1. Effects of CYP2D6 and CYP3A inhibition on the formation of solanidine metabolites. The effects of paroxetine (CYP2D6 inhibitor) and ritonavir (CYP3A inhibitor) on the formation of various solanidine metabolites, including OH-solanidine (m/z 414/98, rt 6.5 min, marked with a star*), from solanidine 5 µM were investigated in HLMs (2 mg/ml) for up to 150 min. CYP, cytochrome P450; HLMs, human liver microsomes. Figure S2. Phase II metabolism of solanidine. UGT-mediated metabolism of solanidine was investigated by incubation of solanidine (0.1 µM) with HLMs (0.2 mg/ml) and UDPGA (a-b) for 30 min, and SULT-mediated metabolism by incubation of solanidine (0.1 µM) with HLC (0.1 mg/ml) and PAPS (c-d) for 30 min. HLC, human liver cytosol; HLMs, human liver microsomes; PAPS, adenosine 3-phosphate 5′-phosphosulfate triethylammonium salt; SULT, sulfotransferase; UDPGA, uridine 5′-diphospho-glucuronic acid; UGT, uridine diphospho-glucuronosyltransferase. Figure S3. Chromatograms of solanidine and its selected CYP2D6 mediated metabolites in plasma. a) Separation of compounds using the chromatographic settings described by Magliocco et al. 2021 and b) using the method presented in this study. Table S1. Demographic characteristics of study subjects. Table S2. CYP2D6 star allele definitions. Table S3. CYP2D6 genotype distribution in the study population. Table S4. Associations of metabolite features found in non-targeted metabolomics analysis of human plasma with CYP2D6 phenotypes. Table S5. Effect of CYP2D6 phenotype on plasma solanidine metabolites in healthy volunteers (n=314). Table S6. Effect of NFIB adjusted CYP2D6 phenotype on plasma solanidine and metabolite to solanidine ratios in healthy volunteers (n=314). Table S7. Solanidine clearance values.