Luna omega 1.6 μm polar c18

PK studies were performed in male CD-1 mice between 18 and 25 g in weight. The test compound was injected intravenously (i.v.) into CD-1 mice at 0.5 mg/kg in formulation containing 5% DMA (dimethyl acetamide) 15% Solutol HS15 and 80% water. Animals were euthanized at predefined time points by CO₂ inhalation and blood was collected in K2-EDTA vacutainers by cardiac puncture. The vials were then centrifuged to collect plasma. The plasma samples were extracted using acetonitrile and then analyzed by LC-MS to determine the concentrations of Compound 1 and Compound 3. LC-MS was performed using a Luna Omega 1.6 μm Polar C18 (Phenomenex) column, a Shimadzu Nexera UHPLC, and a Sciex QTrap 5500 mass spectrometer. The PK parameters were calculated by non-compartmental analysis using Phoenix WinNonlin (Certara).

Compound at 1 μM (0.5% final DMSO concentration) was incubated with a 0.5 mg/mL CD-1 mouse (Sigma), SD rat (Life Technologies), or human liver microsome (Life Technologies) in the presence of 1 mM NADPH at 37°C for up to one hour. Aliquots were taken out at different time points and quenched in acetonitrile containing diclofenac (internal standard). The samples were then centrifuged and the supernatant was analyzed by LC-MS using a Luna Omega 1.6 μm Polar C18 (Phenomenex) column, a Shimadzu Nexera UHPLC, and a Sciex QTrap 5500 mass spectrometer. Verapamil was used as a positive control, and warfarin was used as a negative control.