| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Chicago (Uplift) | A | NT | |
| 2 | Chicago (Ogden International) | A | NT | |
| 3 | Chicago (Amundsen) | A | 49.00 | |
| 4 | Chicago (De La Salle) | A | 45.11 | |
| 5 | ITW David Speer Academy | A | 46.85 | |
| 6 | Chicago (Prosser) | A | NT | |
| 7 | Chicago (Marine Military Academy) | A | NT | |
| 8 | Chicago (Lake View) | A | DQ |