| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | THOMAS, Ava | Nease | 1:05.59 | Tocoi Creek Toro Showdown |
| 2 | PRESCOTT, Kenlyn | Bartram Trail | 1:05.00 | |
| 3 | WILLIAMSON, Kaygen | Clay | 1:04.50 | Bear Classic / True Team Championships |
| 4 | MATOS, Illiana | Creekside | 1:03.82 | Creekside High School Invite 2021 |
| 5 | REICH, Daniela | Nease | 1:04.11 | Tocoi Creek Toro Showdown |
| 6 | CANDLER, Abigail | Columbia | 1:04.51 | Mandarin Mustang Classic |
| 7 | WILLIAMS, Isabella | Bartram Trail | 1:05.49 | Bear Classic / True Team Championships |
| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | LANGLOIS, Lindy | Providence | 1:02.98 | 2019 FHSAA State Track & Field Championships |
| 2 | EPTING, Melissa | Creekside | 1:01.90 | |
| 3 | HAYWARD, Hannah | Providence | 1:00.00 | |
| 4 | DIXON, Skyy | Providence | 57.69 | FHSAA Class 1A Track & Field State Championships |
| 5 | RICE, Caroline | Creekside | 58.53 | Mandarin Mustang Classic |
| 6 | JOHNSON, Ava | St. Johns Country Day | 1:01.81 | Embry-Riddle Spikes Classic - high school |
| 7 | CHIQOUNIS, Mia | Ponte Vedra | 1:02.00 |