| Lane | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|
| 1 | ECCLES, Cassidy | SO | Alton (Marquette) | NT | 1:05.00 |
| 2 | WEDEKEMPER, Lily | FR | Centralia (Christ Our Rock Lutheran) | 1:01.64 | |
| 3 | HENTRICH, Sammy | SR | Alton (Marquette) | 52.31 | |
| 4 | WATKINS, Zoey | FR | Bunker Hill | 53.50 | |
| 5 | HARRIS, Isabella | SO | Glen Carbon (Father McGivney) | 1:02.19 |