| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Riverview |
A | NT | |
| 3 | Leto |
A | NT | |
| 4 | Winthrop College Prep | A | NT | |
| 5 | Spoto |
A | NT | |
| 6 | Lennard |
A | NT | |
| 7 | Freedom (Tampa) |
A | NT |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Wiregrass Ranch |
A | 1:05.55 | |
| 2 | Jefferson |
A | 53.72 | 53.72 George Steinbrenner Senior Night |
| 3 | Gaither |
A | DNS | Western Conference American Division Championships |
| 4 | Armwood |
A | 51.90 | |
| 5 | Palm Harbor University |
A | 52.44 | FHSAA 4A District 6 |
| 6 | Brandon |
A | 53.39 | 53.39 WC Federal Division |
| 7 | Middleton |
A | DNS | 55.39 WC National Division Meet |
| 8 | Strawberry Crest |
A | NT |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Newsome |
A | DNS | |
| 2 | Alonso |
A | 50.41 | FHSAA 4A District 6 |
| 3 | Hillsborough |
A | 49.20 | |
| 4 | Chamberlain |
A | 42.25 | |
| 5 | Tampa Bay Tech |
A | 48.40 | FHSAA Class 4A Track & Field State Championships |
| 6 | Bloomingdale |
A | 50.23 | 50.23 WC Federal Division |
| 7 | Sumner |
A | 51.50 | |
| 8 | Plant |
A | 51.88 | 51.88 George Steinbrenner Invitational |