mm DV; 2 mm × 2 mm). Protein content in punches from the hippocampus, prefrontal cortex, and nucleus accumbens averaged 25–30 μg/sample. Punches from the piriform cortex averaged 10–15 μg protein/sample. Lipids were extracted with chloroform (2 vol) and washed with water (1 vol). Endocannabinoids were fractionated by open-bed silica gel column chromatography, as previously described (Moise et al., 2008, Astarita and Piomelli, 2009). Lipids were reconstituted in chloroform, loaded onto small glass columns packed with Silica Gel G (60-Å 230–400 Mesh ASTM; Whatman, Clifton, NJ, USA), and washed with 2 ml of chloroform. Anandamide and 2-AG were eluted with 1 ml of chloroform/methanol (9:1, vol/vol). Eluates were dried under N2 and reconstituted in 50 μL of methanol for LC/MS analyses.
