Identification and quantification of anandamide and 2-AG was performed using an Applied Biosystems 3200 Q trap with a turbo V source for TurbolonSpray attached to a Shimadzu SCL HPLC system controlled by Analyst 1.4.2 software. A five point calibration curves with concentration ranges of 0.078 to 20 pmol of anandamide and 0.125 to 40 nmols of 2-AG, a blank with and without the internal standards were prepared with each sample batch. A linear regression of the ratio of the peak area counts of anandamide, 2-AG to their corresponding internal standards (anandamide-d8 and 2-AG-d8) versus concentration was used to construct the calibration curves. The lower limit of quantification and lower limit of detection for anandamide were 0.078 pmol and 0.020 pmol, respectively. The lower limit of quantification and the lower limit of detection for 2-AG were 0.125 nmol and 0.0.025 nmol, respectively. All calibration concentrations were calculated to be within 15% of their expected values, except the lower limit of quantification which was within 20 % of its expected concentrations. The lower limit of quantification also had a response at least
