Healthy volunteer samples
Venous blood was collected from three healthy Thai volunteers given a
single dose of IVM (400 µg/kg) (NCT02568098) (Kobylinski et al., 2020).
The blood samples were collected in sodium heparin tubes at 24 hours
post IVM ingestion and kept frozen at -80°C until LC-MS/MS analysis. For
metabolite extraction, whole blood samples were thawed at ambient
temperature, vortexed briefly, and centrifuged at 3000×g for 5
min at 20°C. 50 µL of plasma was transferred into a new microcentrifuge
tube and mixed with 200 µL of cold acetonitrile. Tubes were vortexed for
10 minutes at ambient temperature and centrifuged at 1100×g for 5
min at 20°C. The supernatant was collected, evaporated in speed vacuum,
and kept at -80°C freezer until LC-MS/MS analysis.