In the medieval-themed clip, a king declares to his subjects Bud Light does not use the ingredient.
The advert pointedly refers to rival companies Miller Lite and Coors Light, which both use corn syrup.
"To be clear, Bud Light is not brewed with corn syrup, and Miller Lite and Coors Light are," Bud Light added on Twitter.
High-fructose corn syrup (HFCS) is widely used as an artificial sweetener in the US food industry and is made from corn starch.
Multiple studies have linked the ingredient to a rise in obesity in the US.
Bud Light's latest advert also continues its campaign to become fully transparent about the ingredients used in its products.
But the new advert proved to be a source of irritation within the US corn lobby.
"America’s corn farmers are disappointed in you," the National Corn Growers Association (NCGA) said online, shortly after the Super Bowl advert aired.
"We would love to discuss with you the many benefits of corn!"
Kevin Ross, a Twitter user who appears to be a farmer from Iowa, also described himself as "thoroughly disappointed" by the advert and posted a clip of himself pouring Bud Light down the sink.
"We couldn't say it better," the NCGA wrote.
"The fact that Bud Light isn’t made with corn isn’t an issue. Most of us already knew that," said Matt Boucher, a farmer from Illinois.
"It’s the fact that Bud Light, and now MillerCoors advertised it in a way that insinuates that corn is bad when in fact it isn’t."
— Kevin Ross (@krossfarms)
Miller Lite's social media team also responded to the advert on Twitter.
"Hey Bud Light, thanks for including us in our first Super Bowl ad in over 20 years," the brand said.
"You forgot two things though... we have more taste and half the carbs!"
Bud Light aired other Middle Ages-inspired adverts during the Super Bowl, including one that drew inspiration from Game of Thrones.