Blood samples were collected from the lateral tail vein 30 minutes after the presentation of the bottles in animals that had established stable baseline ethanol consumption. Whole blood was centrifuged at 4°C for 13 minutes at 8000 rpm and sera was separated and stored at −80°C until analysis. Sera was then analyzed using the nicotinamide adenine dinucleotide (NAD)-alcohol dehydrogenase (ADH) spectrophotometric assay (Zapata et al., 2006). The BECs were determined using a standard calibration curve and then correlated with ethanol consumed (g/kg/30min).
