| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Belleville (Althoff Catholic) | A | Thorpe | Cunningham | Hebel | Distler | 55.84 | |
| 4 | Breese (Mater Dei) | A | Kampwerth | Zurliene | Strubhart | Becker | 55.20 | |
| 5 | Freeburg | A | Trentman | Geluck | Trentman | Harmon | DQ | |
| 6 | O'Fallon (First Baptist Academy) | A | Nava | Hagadorn | Finley | Nava | 59.38 |