| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | VALENTIN, Ariyah | Sebring |
2:45.00 | |
| 2 | HERRERA, Saydee | Hardee |
2:45.34 | Avon Park Meet #1 2025 |
| 3 | ALAMIA, Ezmeralda | Hardee |
2:55.95 | Lakewood Ranch Invitational 2025 |
| 4 | TREJO-BALDERAS, Camila | DeSoto County |
3:02.64 | Avon Park Meet #1 2025 |
| 5 | GIORDANO, Olivia | Sebring |
3:07.11 | Avon Park Meet #1 2025 |
| 6 | BIRT, Kaylynn | Sebring |
3:14.58 | Avon Park Meet #1 2025 |
| 7 | DELGADO, Marlene | Avon Park |
3:20.98 | Lake Placid Invite |
| 8 | DIAZ, Ashley | Lake Placid |
3:29.81 | Avon Park Meet #1 2025 |
| 9 | GARCIA, Araceli | Avon Park |
3:34.01 | Avon Park Meet #1 2025 |
| 10 | GREIDER, Caroline | Lake Placid |
3:35.02 | Avon Park Meet #1 2025 |
| 11 | LOPEZ-FERNANDO, Carina | Avon Park |
3:52.59 | Frostproof - The Ridge Open 2025 |
| 12 | DEWAIN, Emily | Avon Park |
3:57.68 | Avon Park Meet #1 2025 |
| 13 | ALVARENGA-RODRIGUEZ, Leslie | Hardee |
4:10.13 | Bob English Invitational 2025 |
| 14 | LOPEZ-CRUZ, Marisol | Hardee |
4:14.53 | Avon Park Meet #1 2025 |
| 15 | AVILA, Jasmin | Lake Placid |
NT | |
| 16 | FRAZIER, Meredith | Sebring |
NT | |
| 17 | HERNANDEZ, Mia | Lake Placid |
NT |