| Lane | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|
| 2 | HOLIWAY, Eleanor | SO | Smith-Cotton High School | 1:04.04 | Verified |
| 3 | HOLLOWAY, Rebecca | FR | Jefferson City High School | 58.22 | Verified |
| 4 | DELK, Lily | SR | Helias Catholic High School | 53.30 | Verified |
| 5 | HOFEDITZ, Riley | FR | Hickman High School | 57.42 | Verified |
| 6 | MILLER, Elizabeth | FR | Jefferson City High School | 1:00.62 | Verified |