Barkley suffered the injury in the second quarter after he fell awkwardly on a six-yard reception near the Giants' sideline. He was helped to the locker room by two team trainers.
Barkley had eight carries for 10 yards before his exit. He added four receptions for 27 yards on five targets.
The 2018 first-round pick also missed a few plays earlier in the game while being evaluated in the medical tent after a hit to the helmet.
Wayne Gallman is expected to step in for Barkley in his absence. The Giants trail the Buccaneers 28-10 at halftime.