| Lane | Athlete | Yr. | Team | Seed |
|---|---|---|---|---|
| 2 | Grace VAHLKAMP | JR | Freeburg | 1:04.90 |
| 3 | Sammy HENTRICH | SR | Alton (Marquette) | 59.40 |
| 4 | Gina TRUAX | FR | Wood River (East Alton-W.R.) | 49.27 |
| 5 | Jayden LUNA | SO | Lebanon | 58.50 |
| 6 | Addison JUNG | FR | Freeburg | 1:03.00 |
| 7 | Jada PARKER | JR | East St. Louis (SIUE Charter | 1:07.80 |