Canada is one of the world’s top producers of mustard, and according to the Canadian Grain Commission, its yield varied between 50,000-286,700 tonnes over the last twenty years. Besides being high in protein and oil, mustard is rich in phenolic compounds with great potential for application in food production, nutraceuticals, and cosmetics. The most noticeable compounds that are found in mustard are sinapine, sinapic acid and its conversion to canolol. However, the production of mustard has not fully exploited this oilseed and its functional potential, especially its bioactivities. Therefore, a more effective extraction method has been researched. In this study, a home-scale system with the application of temperatures, pressure and pH was designed to optimize the extraction of the sinapine, and sinapic acid for the generation of canolol from Oriental, black and yellow mustard varieties. Experiments proceeded with whole and crushed seeds using sautéing as a preheating treatment, followed by acidified-, neutralized- and alkaline-pressurized wet extraction. HPLC analysis, different antioxidant assays together with total phenolic content (TPC) and total flavonoid content (TFC) as long as the anti-tyrosinase activity was used for the quantification of extraction efficiency. This extraction system proved to be productive, with the highest targeted major sinapates obtained from yellow and black mustard. Moreover, the strongest antioxidant and anti-tyrosinase activity was also observed for both yellow and black mustard extracts providing in-vitro evidence for the potential application of mustard for nutraceuticals and in cosmetic production.