| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 2 | Freedom (Tampa) |
A | Hernandez | Daniels | Kolar | Wood | 55.39 |
| 3 | Wharton |
A | Grady | Stegeman | Hurst | Polk | 54.36 |
| 4 | Wharton |
B | Canady | Johnson | Estaban | Jones | 54.36 |
| 5 | Wharton |
C | Bennett | Robinson Bey | Hannah | James | 54.36 |
| 6 | Wharton |
D | Cox | Trought | Marte | Laurent | 54.36 |
| 7 | Freedom (Tampa) |
B | Jackson | Tate | Paulino | Edwards-Ricketts | 55.39 |