| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Pine School |
A | Albury | Leighty | Murray | Sheehan | 48.40 | Jensen Beach Invitational 2026 |
| 3 | Clewiston |
A | Moody | James | Bryant | Ward | DNS | Sebring Meet #2 2025 |
| 4 | Jensen Beach |
A | Sullivan | Fell | Chrysler | Cewe | 42.05 | TFRRS 03/07/25 - 46.26 |
| 5 | Benjamin School |
A | Bowens | Cantave | Douglas | Noble | 42.82 | FHSAA 1A Region 4 |
| 6 | Martin County |
A | Dennis | McLean | Ocampo | Fucile | 44.95 | Jensen Beach Invitational 2026 |