| Lane | Team | Squad | Seed | Note | 
|---|---|---|---|---|
| 1 | Timber Creek | A | NT | |
| 2 | Apopka | A | DNS | |
| 3 | First Academy (Orlando) | A | 4:26.57 | St. Cloud Invitational | 
| 4 | East Ridge | A | 4:14.90 | 4:14.90 Dr. Phillips Panther Invite | 
| 5 | Cypress Creek | A | DNS | |
| 6 | Freedom (Orlando) | A | 4:26.60 | Early Hawk Invitational | 
| 7 | Foundation Academy | A | 4:53.09 | 4:53.09 Windermere Meet of Champions Classic 2022 | 
| Lane | Team | Squad | Seed | Note | 
|---|---|---|---|---|
| 1 | Olympia | A | 4:10.00 | |
| 2 | Jones | A | 4:08.43 | FHSAA 3A Region 2 | 
| 3 | Winter Park | A | 4:04.50 | |
| 4 | Maclay | A | 4:02.64 | FHSAA Class 1A Track & Field State Championships | 
| 5 | South Lake | A | 4:03.38 | FHSAA Class 3A Track & Field State Championships | 
| 6 | Ocoee | A | 4:08.00 | |
| 7 | West Orange | A | DNS | |
| 8 | Boone | A | 4:12.25 |