| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Glades Central |
A | Pinto | Forbes | Blaise | Stewart | 52.53 | FHSAA 2A District 13 |
| 4 | Pahokee |
A | Jones | Johnson | White | Washington | 49.42 | FHSAA 1A Region 4 |
| 5 | Somerset College Prep | A | Bourne-Morgan | Wright | Lopez | Chase | 52.45 | Falcon Classic 2024 |
| 6 | Glades Day |
A | Alvarez | Baez | Sturgis | Bailey | 54.35 | Glade Central Invitational 2024 |