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Gatorade was formulated in 1965 by a team of scientists at the University of Florida College of Medicine, including Robert Cade, Dana Shires, Harry James Free and Alejandro de Quesada. Following a request from Florida Gators football head coach Ray Graves, Gatorade was created to help athletes by acting as a replacement for body fluids lost during physical exertion. The earliest versions of the beverage consisted of a mixture of water, sodium, sugar, potassium, phosphate, and lemon juice.[8] Ten players on the University of Florida football team tested the first version of Gatorade during practices and games in 1965, and the tests were deemed successful. On the other hand, star quarterback Steve Spurrier demurred, "I don’t have any answer for whether the Gatorade helped us be a better second-half team or not. . . . We drank it, but whether it helped us in the second half, who knows?"[9] Nonetheless, the football team credited Gatorade as having contributed to their first Orange Bowl win over the Georgia Tech Yellow Jackets in 1967, at which point the drink gained traction within the athletic community. Yellow Jackets coach Bobby Dodd, when asked why his team lost, replied: "We didn't have Gatorade. That made the difference."

The University of Florida researchers initially considered naming their product "Gator-Aid." They settled on the name Gatorade, however, since the researchers wanted to create a commercial product, not a scientifically-validated one. Darren Rovell notes his history of Gatorade, First in Thirst, "the doctors realized that they probably shouldn't use the 'Aid' suffix, since that would mean that if the drink were ever marketed, they would have to prove that it had a clear medicinal use and perform clinical tests on thousands of people."[11] Gatorade co-inventor Dana Shires explained, "We were told that you couldn't use that because the Food and Drug Administration prohibited that. That would classify it as something other than a cola or soft drink, so we changed it to ade."[12]