| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Avon Park |
A | Jordan | Wilson | Burgess Jr. | Timmons | NT | |
| 3 | Frostproof |
A | Griffiths | Ramirez | Ramirez | Shields | 3:54.00 | |
| 4 | Ridge Community |
A | Davis | Desrosiers | Talabert | Patino | 3:27.45 | FHSAA 4A Region 2 |
| 5 | Tenoroc |
A | Adams | Eugene | Manning | Thomas | 3:46.88 | Blue Devil Invitational (Regio |
| 6 | McLaughlin | A | Scarlett | Dzuray | Gidarisingh | Searight | NT |