| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Bradenton Christian |
A | NT | |
| 3 | Sarasota Military Academy |
A | 56.17 | Larry Richardson Memorial Invitational |
| 4 | Cardinal Mooney |
A | 49.58 | Larry Richardson Memorial Invitational |
| 5 | Winthrop College Prep | A | 55.26 | Blue Devil Invitational (Regio |
| 6 | Out-of-Door |
A | 56.52 | Saint Stephens Falcon Invite 2025 |