| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Tocoi Creek | B | NT | |
| 2 | Tocoi Creek | A | 44.23 | Tocoi Creek Toro Showdown |
| 3 | Fleming Island | A | 43.31 | Danny Brown Invitational |
| 4 | Orange Park | A | 42.00 | FHSAA 3A Region 1 |
| 5 | Creekside | A | 42.82 | FHSAA 4A Region 1 |
| 6 | St. Augustine | A | 43.38 | Tocoi Creek Toro Showdown |
| 7 | Ponte Vedra | A | 43.78 | East Coast Classic |