H1 hybrid multi mode reader plate

Eighteen 14-week old C57BL/6J male mice (Jackson Laboratories) housed in temperature, light, and humidity-controlled conditions with a 12-h light/dark cycle were separated into three groups and administered a single injection as follows: 1) 3.5 g/kg ethanol i.p, injection 2) 3.5 g/kg ethanol + DHM (5 mg/kg) i.p. injection, and 3) 3.5 g/kg ethanol + DHM ( 10 mg/kg) i.p. injection (6 mice per group). Mice were administered ethanol via i.p. injection to ensure constant concentrations of ethanol in all mice for accurate comparisons between groups. All mice were euthanized via CO2 and cervical dislocation 45 minutes post-injection, and whole blood samples were collected, stored at room temperature for thirty minutes, and separated by refrigerated centrifugation for 10 minutes at 2,000 × g. All samples were stored on ice immediately after separation. Serum samples were analyzed immediately after separation using the Ethanol and Acetaldehyde Assay Kit (Megazyme, Bray, Ireland) and H1 Hybrid Multi-Mode Reader Plate (BioTek, Winooski, VT) according to the manufacturer’s guidelines. All acetaldehyde measurements were conducted on ice to keep samples cold and preserve acetaldehyde concentrations in solution.