| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Auburndale | A | NT | |
| 3 | Mulberry | A | DQ | |
| 4 | Bartow | A | 52.95 | George Jenkins - Bob English Invitational 2023 |
| 5 | Haines City | A | 53.54 | |
| 6 | McLaughlin | A | NT | |
| 7 | Lake Region | A | NT |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Fort Meade | A | DNS | FHSAA 1A Region 3 |
| 2 | Davenport | A | 51.52 | Poinciana Relays |
| 3 | Winter Haven | A | 49.82 | 49.82 Kowboy Invitational |
| 4 | Lake Wales | A | 48.32 | Berkley Track and Field Invitational |
| 5 | George Jenkins | A | 48.94 | FSU Relays (High Schools) |
| 6 | Lakeland | A | DNS | FHSAA 3A Region 2 |
| 7 | Lake Gibson | A | DNF | Davenport invitational 2023 |
| 8 | Frostproof | A | 52.46 | Bulldog Invitational 2023 |