| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Out-of-Door |
A | DNS | |
| 3 | Cardinal Mooney |
A | 4:25.24 | FHSAA 2A District 10 |
| 4 | Braden River |
A | 4:30.00 |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Venice |
A | 4:18.30 | |
| 2 | Sarasota |
A | 4:14.13 | FHSAA 4A Region 3 |
| 3 | Riverview (Sarasota) |
A | 4:09.53 | 4:09.53 Wally Keller Invitational |
| 4 | Palmetto |
A | 4:12.65 | FHSAA 4A District 11 |
| 5 | Parrish |
A | 4:15.90 | FHSAA 4A District 11 |
| 6 | North Port |
A | 4:23.60 |