| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | St. Edward's |
A | Ciceron | Cannon | Williams | Simmonds | 49.20 | Viera High School Invitational |
| 4 | Treasure Coast | A | Estassin | Song | Austin | Diejuste | 43.92 | SLC Championships 2025 |
| 5 | Fort Pierce Westwood |
A | Demery | Greer | Lynch | Martin | 44.04 | Cobra Relays 2025 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | John Carroll |
A | Coleman | Pittman | Summerall | NORRIS | 43.67 | Jensen Beach Invitational 2026 |
| 3 | Vero Beach |
A | Dillard | Cabrera | White | Jacobs | 42.47 | FHSAA 4A District 8 |
| 4 | Centennial |
B | Thomas | Johnson | Raphael | Jefferson | 40.96 | Bill Wilson Invitational 2025 |
| 5 | Centennial |
A | Rowley | Hylton | Johnson | Benito | 40.96 | Bill Wilson Invitational 2025 |
| 6 | Benjamin School |
A | Etienne | Reddick | Ferguson | Dalton | 42.82 | FHSAA 1A Region 4 |
| 7 | Tradition Prep | A | Monaco | Joseph | Lopez | Heredia | 43.87 | Bulldog Invitational 2025 |