| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Florida Prep | B | DNS | |
| 2 | Lake Region | A | NT | |
| 3 | Auburndale | A | 4:27.52 | FHSAA 3A Region 2 |
| 4 | George Jenkins | A | 3:57.75 | FHSAA Class 4A & Para Track & Field State Championships |
| 5 | Lake Wales | A | 4:01.47 | FHSAA 3A Region 2 |
| 6 | Haines City | A | 4:39.40 | 4:39.40 Bob English Relays 2022 |
| 7 | Florida Prep | A | DNS | |
| 8 | Haines City | B | 4:50.00 |