Pure compounds belonging to flavonoid family were studied for their biological potential such as rutin hydrate (3, 3′, 4′, 5, 7-pentahydroxyflavone-3-rhamnoglucoside). These molecules were chosen based on their neuroactive properties. The evaluation of the in vitro de-addiction activity were exploited for the first time for such purposes. The aim of the present study was to investigate the effects of rutin hydrate (3, 3′, 4′, 5, 7-pentahydroxyflavone-3-rhamnoglucoside), a flavonoid that is an important dietary constituent of foods and plant-based beverages, on response-reinstatement of mice using running alley method of ethanol reward. Rutin (0.1, 1, and 10 mg/kg) was administered orally (p.o) one hour before post conditioning test, during extinction trials, and on re-instatement day. It was noted that rutin significantly (P < 0.001) increased runtimes on reinstatement day. No changes in body weight, food and water intake when compared with saline control group was observed. These results indicate that flavonoid rutin hydrate has a potential role in decreasing ethanol reinforcement. This can be used as a powerful food to delineate ethanol re-inforced running. Several mechanisms may contribute to the potential role of rutin in ethanol re-inforced running. Therefore, further studies are warranted to understand the exact mechanism of rutin hydrate.