| Lane | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|
| 2 | Stevanovich, Alex | JR | Chicago (University) | 1:02.31 | |
| 3 | O'Toole, Charlotte | FR | Chicago (Latin) | 59.11 | |
| 4 | Steele, Thandi | SR | Winnetka (North Shore Country Day) | 56.80 | |
| 5 | Nappo, Sophia | JR | Chicago (Latin) | 59.11 | |
| 6 | Valliath, Maya | JR | Winnetka (North Shore Country Day) | 1:01.16 |