| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Braden River |
A | DQ | |
| 3 | Palmetto |
A | 51.42 | 51.42 Palmetto High Invitational |
| 4 | Sarasota |
A | 54.29 | 54.29 Sarasota Riverview - Ram Invite 2025 |
| 5 | Out-of-Door |
A | DNS |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Venice |
A | 50.90 | |
| 2 | Riverview (Sarasota) |
A | 50.15 | FHSAA 4A Region 3 |
| 3 | Cardinal Mooney |
A | DQ | 49.58 Larry Richardson Memorial Invitational |
| 4 | Parrish |
A | 50.00 | |
| 5 | North Port |
A | 50.20 | |
| 6 | Clearwater Central Catholic |
A | 51.39 | Keswick Crusader Invitational |