Dysmenorrhea (menstrual cramps) is defined as difficult menstrual flow or painful menstruation and the most important cause of chronic pelvic pain. It caused by an increase in prostaglandin which has an inflammatory and painful effect. Flavonoids are constituents that can inhibit the formation of prostaglandins. In this study, extraction of flavonoid compounds from mother in law's tongue (Sansevieria trifasciata) leaves was evaluated using Microwave-Assisted Enzymatic Extraction (MAEE) method. The optimal extraction conditions were as follows: enzyme concentration 40 mg/100 mL distilled water and 70% ethanol concentration. Under these conditions, the extraction yield of S. trifasciata achieved was 42.53%, which represented an increase of 83%, respectively, compared with the control experiment (23.27%). The flavonoid content at optimum MAEE conditions reached 0.52 mg/g extract, while the control only obtained 0.32 mg/g extract. Anti-inflammatory activity test of mother in law's tongue leaves was done using in vivo method in white male rats (Rattus norvegicus). The inhibition percentage showed that S. trifasciata extract (500 mg/kg BB) was potential to inhibit anti-inflammatory activity by 70.32% relative to Na Diclofenac as a positive control. This results concluded that MAEE methods was an effective extraction method to improved extraction yield and flavonoid content of S. trifasciata which can be used for anti-inflammatory conditions such as menstrual cramps.