Pancreatin 4 usp

All compounds were purchased from Sigma-Aldrich (St. Louis, Mo, USA) except for mefloquine (Roche, Mannheim, Germany) and piperaquine (Sigma-tau, Rome, Italy). Stock solution of chloroquine, amodiaquine, piperaquine and tetracycline were dissolved in water; quinine in DMSO; mefloquine, halofantrine, artemisinin, artesunate in methanol; and rolytetracycline and doxycycline in ethanol 70 % and pyrimethamine in absolute ethanol. Based on the investigation of growth conditions, these compounds were tested with: (1) the baseline medium, (2) blood extract substituted with haemin at 5 mM in DMSO, (3) pancreatin 4× USP (Sigma-Aldrich) at 10 % and (4) 2.5 %. A range of doubling concentrations from 15.6 to 1000 µM was tested under these four conditions. After preparing the seeded mediums with the compounds serial dilutions, were incubated for 5–7 days at 37 °C in 5 % CO₂. The plates were observed daily. The results from the 48 h incubation time points were used to establish inhibition.

β-carotene (BC) (>93%, UV), sodium caseinate (SCN), Tween^® 80 (Polysorbate 80, TW80), pepsin (≥250 unit/mg), pancreatin (4× USP), bile salts, Dulbecco’s Modified Eagle’s Medium (DMEM) (containing 4.5 g/L D-glucose), penicillin and streptomycin (100×), fetal bovine serum (FBS), phosphate-buffered solution (PBS) and cell lysis buffer were purchased from Sigma-Aldrich (St. Louis, MO, USA). Sunflower oil was purchased from a local supermarket, and whey protein isolate (WPI) was obtained from Davisco Food International (Le Sueur, MN, USA). All other chemicals and reagents used were of AR-grade and obtained from Sigma-Aldrich.