| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | North Port |
A | 48.00 | |
| 3 | Braden River |
A | 46.39 | 46.39 Manatee County Track and Field Championship |
| 4 | Parrish |
A | 47.96 | Spring Break Invitational 2025 |
| 5 | Out-of-Door |
A | DNS |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Venice |
A | 43.90 | |
| 2 | Clearwater Central Catholic |
A | 43.11 | 43.11 Mustang Invitational |
| 3 | Cardinal Mooney |
A | 42.12 | Guy Thomas Memorial -Sponsored by Anderson Electric of SWFL |
| 4 | Palmetto |
A | DNS | |
| 5 | Riverview (Sarasota) |
A | 43.68 | 43.68 2025 Sarasota County Championships |
| 6 | Sarasota |
A | 45.53 | 45.53 Sarasota Riverview - Ram Invite 2025 |