Flavonoids with focus on the rutin can have beneficial effects for human health already scientifically proven due to its antioxidant properties that can be found in the acerola and its processing wastes. Therefore, the compounds extraction yields with high aggregated value from the wastes using various extraction techniques must be studied. The approach adopted in this work was the rutin extraction from acerola wastes using various extraction methods (conventional, assisted by ultrasound and combined). The rutin extractions from the acerola waste in ATPS were able to separate the flavonoid to top and bottom phases. The results of separation coefficient and extraction efficacy show that it is possible to conceive extraction processes that are cheaper and simpler, able to promote the rutin biomolecule separation.