| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Tradition Prep | A | Parrel | Falcon | Castro | Derius | DNS | Cobra Relays 2025 |
| 3 | John Carroll |
A | VEN RENS | Martinez | PEREZ | ANDERSEN | 4:45.47 | Jensen Beach Invite 2025 |
| 4 | Dwyer |
A | Smith | Jean | Flowers | Verdecia Perez | 4:10.78 | FHSAA 4A District 8 |
| 5 | Clewiston |
A | Miscaide | Bradford | Tolbert | Alarcon | DNS | Sebring Meet #2 2025 |
| 6 | Treasure Coast Classical | A | Easom | Conrad | Vaden | Sullivan | 5:14.96 | Jensen Beach Invitational 2025 |