| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Kathleen |
A | DNS | |
| 4 | Davenport |
A | DNS | |
| 5 | Tenoroc |
A | DNS |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Bartow |
A | NT | |
| 2 | Hardee |
A | 4:31.96 | FHSAA 2A Region 3 |
| 3 | McKeel |
A | 4:27.97 | FHSAA 2A Region 3 |
| 4 | Plant City |
A | 4:25.00 | |
| 5 | Lake Region |
A | 4:25.40 | 4:25.40 Lake Region - Thunder Relays2023 |
| 6 | Auburndale |
A | 4:28.71 | 4:28.71 St. Cloud Invitational |
| 7 | Haines City |
A | DNS | 4:55.22 Poinciana Relays |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | East Ridge |
A | 4:18.00 | |
| 2 | First Academy (Orlando) |
A | 4:10.76 | FHSAA Class 1A Track & Field State Championships |
| 3 | Largo |
A | 4:01.07 | 4:01.07 Pinellas County Athletic Conference %28PCAC%29 |
| 4 | George Jenkins |
A | 3:54.93 | FHSAA Class 4A Track & Field State Championships |
| 5 | Hillsborough |
A | 3:58.99 | FHSAA 3A Region 3 |
| 6 | Lake Wales |
A | DNS | |
| 7 | Lake Nona |
A | 4:11.75 | 4:11.75 Falcon Last Chance 13 |
| 8 | Frostproof |
A | DNS | FHSAA 2A Region 3 |