| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Mulberry | A | NT | |
| 4 | Celebration | A | NT | |
| 5 | Vero Beach | A | NT |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | McLaughlin | A | NT | |
| 3 | Santa Fe Catholic | A | NT | |
| 4 | Lake Region | A | DQ | 57.28 Frostproof - Bulldog Invite 2022 |
| 5 | Tampa Bay HEAT | A | 1:02.23 | FHSAA 1A Region 3 |
| 6 | Bell Creek | A | NT | |
| 7 | Bartow | A | NT |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | South Lake | A | 56.00 | |
| 2 | Fort Meade | A | 52.04 | FHSAA 1A Region 3 |
| 3 | Lake Wales | A | 50.03 | FHSAA 3A Region 2 |
| 4 | Niceville | A | 47.34 | FHSAA Class 4A & Para Track & Field State Championships |
| 5 | Plant City | A | 49.00 | |
| 6 | George Jenkins | A | 50.35 | 50.35 6th Annual George Jenkins Invitational 2022 |
| 7 | Frostproof | A | DNS | 52.10 Bob English Relays 2022 |
| 8 | Victory Christian (LL) | A | 56.10 |