Fig. 7.

Degradation of anandamide and 5,6-EET-EA by mouse brain proteins. Anandamide or 5,6-EET-EA (5 μM) were incubated in the presence of mouse brain homogenate (0.5 mg protein/reaction) in phosphate buffer, pH 7.4, at 37°C for 0 to 180 min. At the designated time points and after the addition of internal standard, the reaction mixtures were extracted with 3 volumes of ethyl acetate and analyzed by ESI-LC/MS as described under Materials and Methods. The amounts of 5,6-EET-EA or anandamide remaining at each time point were plotted as a percentage of the starting amount (at time 0). Control samples contained 0.5 mg of heat-inactivated mouse brain homogenate. ^**, p < 0.01; ^***, p < 0.001 compared with the anandamide group at the same time point.