| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Kathleen |
A | NT | |
| 2 | Santa Fe Catholic |
A | 58.48 | 58.48 Lakeland City meet @ George Jenkins 2023 |
| 3 | Hardee |
A | 53.79 | FHSAA 2A Region 3 |
| 4 | Davenport |
A | 51.26 | 51.26 Polk County Championship 2023 |
| 5 | Haines City |
A | DNS | 53.08 Polk County Championship 2023 |
| 6 | East Ridge |
A | DNS | |
| 7 | Bartow |
A | NT | |
| 8 | Lake Region |
A | NT |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Plant City |
A | 51.00 | |
| 2 | Lakeland |
A | 50.28 | FHSAA 3A Region 2 |
| 3 | George Jenkins |
A | 49.14 | |
| 4 | Largo |
A | 47.48 | FHSAA Class 4A Track & Field State Championships |
| 5 | Lake Wales |
A | DQ | |
| 6 | Hillsborough |
A | 49.20 | |
| 7 | First Academy (Orlando) |
A | 50.86 | FHSAA 1A Region 2 |
| 8 | Frostproof |
A | DNS | 51.25 Polk County Championship 2023 |