| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 2 | BARAYBAR, Ariela | Cambridge Christian |
NT | NT |
| 3 | RODRIGUEZ, Arie | Calvary Christian (PtStL) | NT | NT |
| 4 | WALKER, Ava | Classical Academy (Sarasota) |
33.19 | Hurricane Invitational Track and Field Meet |
| 5 | RAMIREZ, Kaela | Cambridge Christian |
34.42 | East Lake Early Bird Invitational |
| 6 | GIBSON, Avery | Bradenton Christian |
NT | NT |
| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 2 | CHIDESTER, Kendal | Sarasota Christian | 32.28 | Larry Richardson Memorial Invitational |
| 3 | GUZMAN, Grace | Duval Charter Baymeadows |
30.86 | Panther Invite at Ridgeview High School |
| 4 | VENTRESS, Andrea | Duval Charter Baymeadows |
30.66 | Tocoi Creek Toro Showdown 2025 (Approved Qualifier) |
| 5 | GONZALEZ, Nastassia | Calvary Christian (PtStL) | 30.74 | Jensen Beach Invitational 2025 |
| 6 | BELL, Lola | Bradenton Christian |
30.88 | Hurricane Invitational Track and Field Meet |
| 7 | LEYVA, Madison | Master's Academy (VB) |
32.55 | Pineapple Cove MS Track 1 2025 |
| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | PIERRE, Niya | Bradenton Christian |
29.22 | John Tasopolos Memorial Invitational |
| 2 | BROWN, Kendyl | Cambridge Christian |
29.14 | Mustang Invitational |
| 3 | TRESSELL, Bryhanna | Babcock | 28.66 | 65th Annual Edison Relays |
| 4 | ANDERSON, Akeeylah | Duval Charter Baymeadows |
27.73 | Tocoi Creek Toro Showdown 2025 (Approved Qualifier) |
| 5 | GOMEZ, Yanela | Babcock | 28.04 | Saint Stephens Falcon Invite 2025 |
| 6 | HOFFMIN, Kathryn | Babcock | 28.83 | Saint Stephens Falcon Invite 2025 |
| 7 | MCMILLEN, Aivly | Indian Rocks Christian |
29.16 | Shorecrest Senior Night |
| 8 | LUBKE, Elliana | Indian Rocks Christian |
30.60 | Calvary Christian Warrior Invitational |