| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Civic Memorial | A | Crymes | Shields | Govero | Brousseau | 55.52 | |
| 3 | Alton Marquette | A | Cain | Sechrest | Rae | Wells | 54.00 | |
| 4 | Madison | A | Gregory | Kelly | Belford | Rosser | 52.02 | |
| 5 | Roxana | A | Lohman | Carlisle | Eads | Daugherty | 53.99 | |
| 6 | East Alton-Wood River | A | Brown | Minner | Nguyen | Truax | 55.10 | |
| 7 | Father McGivney Catholic | A | Harris | Ivnik | Windorf | Pinkston | 59.00 |