| Athlete | Team | Seed | |
|---|---|---|---|
| 1 | Smith, Payton | Avon Grove | 48.52 |
| 2 | Eberly, Lorelei | Coatesville | 49.20 |
| 3 | Ericson, Rylee | D-East | 50.37 |
| 4 | Hearn, Ryleigh | D-West | 51.50 |
| 5 | Compher, Emma | D-West | 52.16 |
| 6 | Baumann, Chloe | D-East | 54.55 |
| 7 | Caronia, Alex | WC Henderson | 55.79 |
| 8 | Jennings-Scott, Giovanna | D-East | 56.24 |
| 9 | Brunschwyler, Charlotte | D-West | 57.00 |
| 10 | Hauer, Carly | D-East | 57.28 |
| 11 | Shaefer, Reilly | D-East | 57.36 |
| 12 | Nolan, Zoe | WC Henderson | 58.04 |
| 13 | Uhlman, Deborah | Bishop Shanahan | 58.37 |
| 14 | Yarnall-Stevenson, Charlotte | D-East | 58.57 |
| 15 | van der Steur, Anastasia | Bishop Shanahan | 58.61 |
| 16 | Montgomery, Alexis | D-West | 58.79 |
| 17 | Hangey, Rebecca | Bishop Shanahan | 59.00 |
| 18 | Fuhr, Mya | D-West | 59.22 |
| 19 | Sujesh, Keerthana | D-East | 59.57 |
| 20 | Davies, Grace | D-East | 1:00.82 |
| 21 | Keefe, Maeve | WC East | 1:01.00 |
| 22 | Carlow, Ana | WC Henderson | 1:02.20 |
| 23 | Drucis, Anna | WC East | 1:03.00 |
| 24 | Robinson, Julia | D-East | 1:03.76 |
| 25 | Bousquet, Sophie | D-East | 1:08.68 |
| 26 | Conrad, Caroline | WC Henderson | 1:17.07 |
| 27 | Rogelstad, Sarah | Bishop Shanahan | NT |