Introduction: More than 90 countries have given the artificial sweetener aspartame the green light to be used in thousands of food and beverage products. Two hundred times sweeter than sugar, aspartame allows food manufacturers to produce sweet foods they can market as “low calorie,” “diet,” or sugar-free,” appealing to hundreds of millions of consumers looking to cut sugar from their diets. Concern relating to the possible adverse effect has been raised due to aspartames metabolic components. Aspartame is rapidly and completely metabolized in humans and experimental animals to aspartic acid (40%), phenylalanine (50%) and methanol (10%). Methanol, a toxic metabolite is primarily metabolized by oxidation to formaldehyde and then to formate these processes are accompanied by the formation of superoxide anion and hydrogen peroxide. Lacuna and Method: This study focus is to understand whether the oral administration of aspartame (40mg/kg.bw) for 90 days, has any effect on membrane bound ATPases, antioxidant status(both enzymatic and non-enzymatic) in spinal cord and motor behavior of Wistar albino rats. To mimic human methanol metabolism, folate deficient rats were used. Result: After 90 days of aspartame administration, showed a significant alteration in membrane bound ATPases, decrease in both enzymatic and non-enzymatic antioxidant level while there was no significant change was observed in motor behavior. Conclusion: This study concludes that oral administration of aspartame (40mg/kg.bw) for longer duration may cause oxidative stress in spinal cord, which didn’t have any consequence on motor behavior, but which may be the root of other neuronal complication because oxidative stress in spinal cord can’t be ignored.