| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Frostproof |
A | Jones | Tibado | Griffiths | Butler | 57.00 | |
| 3 | Moore Haven |
A | Herrera | Myers | Hughes | Faison | 53.54 | Jensen Beach Invitational 2026 |
| 4 | Avon Park |
A | Glabach | Johnson | McMillan | Destino | 52.48 | Tenoroc High School Invitational at SEU |
| 5 | Sebring |
A | Gray | Greene | Wellington | Madison | 53.35 | Bob English Invitational 2026 |
| 6 | Hardee |
A | Pierre | Manriquez | Cornell | Jackson | 54.00 | |
| 7 | Lake Placid |
A | Dillon | Carlini-Smith | Johnson | Jacobs | NT |