2.6. Determination of In Vivo Anti-Inflammatory Potential

The animal assays were conducted as reported earlier [30] with some modification. Firstly, BALB/c juvenile mice (20–25 g) obtained from the stock at the Bioterium of Faculty of Medicine, University of Chile were used. Subsequently, inflammation was induced in the right ear in the control mouse group (n = 16) by the topical application of 5 μg/ear of phorbol 12-myristate 13-acetate (TPA) or 2 mg of arachidonic acid (AA) dissolved in 20 μL of acetone (negative control = 100% inflammation), while acetone (20 μL) was topically applied to each mouse’s left ear (vehicle). In parallel, extracts reconstituted in acetone (3.0 mg/ear) obtained from dried cauliflower by different methods or indomethacin (1.0 mg/ear; positive control or reference drugs against TPA) or nimesulide (0.5 mg/ear; positive control or reference drugs against AA) were applied to the other groups of mice immediately after inducing right ear inflammation (group treated with anti-inflammatory agents; n = 8). The left ear in this group also received only acetone as vehicle. After 6 h or 1 h of ear edema induction by TPA or AA, respectively, the mice were euthanized by CO₂ asphyxiation, and ear samples (6 mm of tissue from both ears) were collected using a mechanical punch. Edema was measured as the difference in the weights between the punches from right and left ears of the negative control and the treated mouse groups, respectively.