Barkley, who was the second overall pick of the 2018 NFL Draft, watched practice in uniform -- sans pads -- after sustaining a mild strain of his left leg during Monday's session.
The 21-year-old Barkley reeled in a pass down the sideline from quarterback Kyle Lauletta before coming up a bit gimpy.
Giants coach Pat Shurmur said that the team will be "smart with him" as the week progressed.
Barkley lived up to the hype befitting his lofty draft status in New York's preseason opener, rumbling 39 yards on his first carry in a 20-10 loss to the visiting Cleveland Browns. He rushed for 43 yards on the evening.
New York's ground game was a train wreck last season and Barkley was viewed as the perfect fit after amassing 1,271 yards and 18 touchdowns last season for Penn State. He also showed he was a three-down back with 54 receptions.
The 6-foot-3, 233-pound Barkley continued to rocket up draft boards after lighting up the 2018 NFL Combine with a spectacular workout, including a 4.40 40-yard dash and a 41-inch vertical jump.