| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Lake Placid |
A | Johnson | Carlini-Smith | Dillon | Jacobs | 54.69 | Avon Park Red Devil Invitational |
| 4 | Avon Park |
A | Davi | Glabach | Destino | Johnson | 52.26 | Avon Park Red Devil Invitational |
| 5 | Sebring |
A | Gray | Greene | Wellington | Madison | 52.35 | Avon Park Red Devil Invitational |
| 6 | DeSoto County |
A | Johnson | Johnson | King | Hannon | NT |