| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Tenoroc |
A | Jean-Louis | Durr | Fernandez | Wilson | 55.17 | Blue Devil Invitational (Regio |
| 3 | Ridge Community |
A | Duvivier | Jackson | Agenor | Cooper | 53.84 | |
| 4 | Frostproof |
A | Smith | Vasquez | McNeil | Ingraham | 53.51 | 53.51 Davenport - Bronco Invitational 2024 |
| 5 | Avon Park |
A | Davi | Greene | Watt | Glabach | 53.52 | FHSAA 2A Region 3 |
| 6 | McLaughlin | A | Menzie | Owens | Robinson | McGriff | 55.00 |