| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Mainland | A | 55.78 | Spruce Creek Developmental Meet #1 |
| 2 | Seabreeze | A | 55.36 | Volusia Freshmen/Sophomore Meet |
| 3 | Olympia | A | 52.42 | Poinciana Relays |
| 4 | Forest | A | 50.10 | 2019 FHSAA State Track & Field Championships |
| 5 | Spruce Creek | A | 51.66 | East Coast Classic |
| 6 | University (OC) | A | DNS | |
| 7 | Father Lopez | A | DQ | BMCHS Catholic Relays |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Lake Mary | A | NT | |
| 4 | Matanzas | A | 58.50 | |
| 5 | Menendez | A | DQ |