| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 2 | Chaminade College Prep | A | Birchfield, Kylan | Chrismer, Blake | Crawford, Brendan | Taylor, JT | 48.84 |
| 3 | St. Louis University | A | Syberg, Carson | Cummings, Maceo | Utsey, Delton | Wolf, Austin | 47.00 |
| 4 | DeSmet Jesuit | A | Shea, Jackson | Rush, Cameron | Rodgers, Cameron | Fields, Kendall | 45.94 |
| 5 | Christian Brothers College | A | Billingsley, Malik | Bradford, Bryan | McCoy, Tyus | Player, Kameron | 46.54 |
| 6 | St. John Vianney | A | Coleman, Cameron | Foster, Cameron | Laub, Mitchell | Moore, Tyler | 48.34 |