| Lane | # | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|---|
| 3 | 279 | MITCHELL, Jada | FLORIDA EAST COAST SUPERSONICS | NT | ||
| 4 | 173 | CALDERON, Aryana | East Orlando Track Club | 1:04.00 | ||
| 5 | 199 | MCFAYDEN, Nyla | Unattached | 59.00 | ||
| 6 | 7 | CALIX, Selena | Riviera Beach Chargers | 1:03.99 | ||
| 7 | 30 | CLARK, Leigha | Take Flight Athletics | NT | ||
| 8 | 39 | KIRTEN, Andreanna | Take Flight Athletics | NT |