| Lane | # | Athlete | Team | Seed | Note |
|---|---|---|---|---|---|
| 1 | 49 | RODRIGUEZ, Sofia | Wesley Chapel | NT | |
| 2 | 36 | SCHMIDT, Sierra | Wesley Chapel | NT | |
| 3 | 105 | SAINTELMY, Abigail | Wiregrass Ranch | 1:12.00 | |
| 4 | 92 | KHALIL, Maryam | Wiregrass Ranch | 1:00.84 | Berkeley Track and Field Invitational |
| 5 | 86 | CLAUNCH, Anissa | Wiregrass Ranch | 1:07.05 | Berkeley Track and Field Invitational |
| 6 | 89 | GREEN, Jayden | Wiregrass Ranch | 1:25.00 | new |
| 7 | 77 | STEVENS, Savannah | Wesley Chapel | NT | |
| 8 | 25 | SCOTT, Kaylyn | Wesley Chapel | NT |