| Athlete | Yr. | Team | Seed | |
|---|---|---|---|---|
| 1 | Bell, Daunte | JR | PA United | 48.00 |
| 2 | Scott, Mason | JR | PA United | 49.20 |
| 3 | Battle, Davon | 15 | Unattached | 50.76 |
| 4 | Wagner, Benjamin | SR | Unattached | 50.80 |
| 5 | Castelli, Wesley | Unattached | 51.64 | |
| 6 | White, Ryan | SR | Unattached | 52.30 |
| 7 | Grace, Edward | 9 | Unattached | 52.58 |
| 8 | Sloboh, Emmanuel | 9 | Harry S Truman 01 | 52.80 |
| 9 | Lotoza, Will | SR | Unattached | 52.97 |
| 10 | Howey, Jackson | 9 | West Chester Track Club | 53.60 |
| 11 | Kengo, Oumaru | SR | Unattached | 54.00 |
| 12 | White, Jaidyn | JR | Unattached | 54.00 |
| 13 | Vaughn, Frederick | 8 | Unattached | 57.00 |
| 14 | Johnson, Madison | 13 | PA United | 57.95 |
| 15 | Barnes, Satchel | JR | Unattached | 58.08 |
| 16 | Henderson, Amiya | 8 | Athletic Development Group | 58.79 |
| 17 | Savage, Avah | 13 | PA United | 58.95 |
| 18 | Adeleke, Jeremiah | 13 | Unattached | 1:00.32 |
| 19 | Richardson, Spencer | 7 | West Chester Track Club | 1:04.95 |
| 20 | Tinsley, Robin | SR | Unattached | 1:08.00 |
| 21 | Banks, Amira | 9 | Athletic Development Group | 1:08.00 |
| 22 | Soto-Villafane, Jaylin | SO | Unattached | 1:09.89 |
| 23 | Jones, Emma | 13 | Unattached | 1:14.53 |
| 24 | Stulak, Beckham | 5 | West Chester Track Club | 1:27.85 |
| 25 | Powell, Kate | Unattached | NT | |
| 26 | Banks, Emmanuel | 8 | West Chester Track Club | NT |
| 27 | Barkhouse, Wren | 4 | West Chester Track Club | NT |