| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Frostproof |
A | Cobb | Blocker | Andrews | Robey | 53.00 | |
| 3 | Ridge Community |
A | Francis | Dameus | Palmer | Parkes | 44.10 | |
| 4 | Tenoroc |
A | Delu | Jones | Garrett | Furman | 43.43 | FHSAA 2A District 10 |
| 5 | Avon Park |
A | Kirkland | Sholtz | Sheilds | Jordan | 43.63 | FHSAA 2A Region 3 |
| 6 | Mulberry |
A | Grant | Nichols | Sledge | Williams | DNS | FHSAA 2A Region 3 |
| 7 | McLaughlin | A | Scarlett | Gidarisingh | Dzuray | Searight | NT |