| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Pine School |
A | Albury | Anderson | Murray | Sheehan | 48.40 | Jensen Beach Invitational 2026 |
| 3 | American Heritage (DB) |
A | Martin | Kiyota Ayrosa | Wilson | Cesar | 45.05 | Welington Meet #1 |
| 4 | Spanish River |
A | Aidonis | Smalls | Lamas | Miller | DNS | FHSAA 4A District 13 |
| 5 | A | Charlot | McKay | Oscar | Vilma | DQ | FHSAA 3A Region 4 | |
| 6 | Spanish River |
B | Bruno | Rollny | Alexander | Toscano | 45.75 | |
| 7 | American Heritage (DB) |
B | Kobe | Kindred | Joseph | Tyson | 50.00 |