| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Spanish River |
B | Anderson | Gomez | Diaz | Stepien | DQ | |
| 4 | Spanish River |
A | Ulysse | Theissen | Main | Fray | 51.56 | Spanish River HS Invitational Track and Field 2026 |
| 5 | A | Pierre | Dominguez | Massena | Michel | 52.44 | FHSAA 3A Region 4 | |
| 6 | American Heritage (DB) |
A | Imami | Klein | Dorsonne | Morales | DNS |