| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Frostproof |
A | Cobb | Sloan | Andrews | Ramirez | 46.00 | |
| 4 | Hardee |
A | Cherry | Myrick | Brunson | Watson | 45.21 | Larry Richardson Memorial Invitational |
| 5 | Lake Region |
A | Luxe | German | Glanton | Christie | DNS | Blue Devil Invitational (Regional Qualifier) |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Davenport |
A | Snow | Dabrio | Key | Brooks | 44.78 | Blue Devil Invitational (Regio |
| 2 | Haines City |
A | Bordes | Bordes | Browne | Deyant | 43.98 | Kowboy Invitational |
| 3 | Lakeland Christian |
A | Bellamy | Truedell | Goines | Calhoun | DNS | Coyote Invitational Cypress Creek Wesly Chapel |
| 4 | Winter Haven |
A | Alls | Brown | Vildor | Washington | DQ | Blue Devil Invitational (Regional Qualifier) |
| 5 | Tenoroc |
A | Delu | Jones | Garrett | Furman | 43.02 | Lakeland City Championships @ George Jenkins 2025 |
| 6 | Sebring |
A | Shannon | Austin | Walker | Olds | 43.49 | Avon Park Meet #1 2025 |
| 7 | McLaughlin | A | Scarlett | Gidarisingh | Dzuray | Searight | 44.52 | Lake Region Sprint Hurdle Night 2025 |