| Lane | # | Athlete | Team | Seed |
|---|---|---|---|---|
| 2 | 156 | Weber, Sophie | Daniel Boone High School | 55.69 |
| 3 | 75 | Wolfe, Reagan | Dobyns Bennett High School | 54.00 |
| 4 | 312 | Hale, Caroline | Science Hill High School | 50.26 |
| 5 | 20 | Feldmann, Marley | Dobyns Bennett High School | 49.71 |
| 6 | 228 | Brown, Carley | West Ridge High School | 50.04 |
| 7 | 368 | Williams, Ava | University School of Johnson City | 53.06 |
| 8 | 265 | Hale, Audrey | Science Hill High School | 55.00 |
| 9 | 174 | Brown, Tyla | Daniel Boone High School | 56.31 |
| Lane | # | Athlete | Team | Seed |
|---|---|---|---|---|
| 4 | 138 | Clawson, Lexie | Daniel Boone High School | NT |
| 5 | 287 | Emmert, Audrey | Science Hill High School | 57.32 |
| 6 | 134 | Morrow, Jade | Daniel Boone High School | NT |