| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Fenton | A | Kelly, Devamon | Ayala, Eric | Bermudez, Cesar | Cabral, Javier | 48.57 | |
| 3 | Willowbrook | A | Florey, Patrick | Davila, Luis | Smith, Justin | Gholston, D'Onte | 48.25 | |
| 4 | Addison Trail | A | D'Ambrose, Nick | Mosely, Pablo | White, Albert | Serna, Sergio | DNS | |
| 5 | Franklin Park-Northlake (Leyden) | A | Mutute, Andres | Cuturic, Sandin | Mayor, Joseph | Alfano, Sal | 47.50 | |
| 6 | Addison Trail | B | Espinoza, Eddie | Whitaker, Braden | Herrera, Cesar | Rodriguez, Angel | 48.56 | |
| 7 | Willowbrook | B | Rosas, Casey | Aly, Mohammed | Roman, Ismael | Berlin, Sebastian | NT |