| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Spanish River |
B | Vassanji | Borges | Cleary | Gromosaik | 5:00.11 | |
| 3 | B | Lewin | Hollis | Dorvil | Meadows | 4:40.21 | FHSAA 3A District 14 | |
| 4 | American Heritage (DB) |
A | Meran | Morales | Bonaros | Klein | 4:01.37 | PBC Meet 2025 |
| 5 | Spanish River |
A | Popescu | Larancuent | Downes | Ahoeiva | 4:30.55 | |
| 6 | A | Mitchell | Gring | Paciletti | Arce | 4:40.21 | FHSAA 3A District 14 | |
| 7 | Pine School |
A | DeGeorge | Stringfellow | Kaliniak | Klostreich | NT |