| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | Williams, Nathaniel | East Carolina |
NT | |
| 2 | Fleming, Jalen | 50.81 | 4th Annual Adidas Trojan Challenge | |
| 3 | Anderson, Nickolis | 49.31 | 4th Annual Adidas Trojan Challenge | |
| 4 | Cooper, Josh | 48.73 | CAA Outdoor Track and Field Championships | |
| 5 | Wright, Cooper | 49.16 | Wendy's Teal Seahawk Invitational | |
| 6 | Mathews, Blake | East Carolina |
50.05 | NCAT Aggie Invitational |
| 7 | Kobziak, Ryan | 51.17 | 4th Annual Adidas Trojan Challenge |