The robot, dubbed Cassie, took to the track at the school's Whyte Track and Field Center and ran the 100 meters in 24.73 seconds.
Cassie previously managed to run a 5-kilometer (3.1-mile) course in 2021 in just over 53 minutes.
The development team said Cassie is the first bipedal robot to use machine learning to control a running gait on outdoor terrain.
"We have been building the understanding to achieve this world record over the past several years, running a 5k and also going up and down stairs," graduate student Devin Crowley, who led the Guinness effort, said in the news release.
"Machine learning approaches have long been used for pattern recognition, such as image recognition, but generating control behaviors for robots is new and different."