| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Marion County High School | A | Morgan | Walker | Kindhart | Kenney | 1:00.63 | Verified |
| 2 | South Shelby High School | A | Bowen | Gill | Magruder | Roush | 52.00 | Unverified: |
| 3 | Palmyra High School | A | King | Kroeger | Zeiger | Juarez | 49.66 | Verified |
| 4 | Clark County High School | A | Fox | Rodgers | CHURCH | Morss | 46.55 | Verified |
| 5 | Kirksville High School | A | Ingersoll | Williams | Dierling | Crawford | 48.22 | Verified |
| 6 | Monroe City High School | A | Hathaway | Reinberg | Walkup | Walters | 51.56 | Verified |
| 7 | Highland High School | A | Dare | Walton | Geisendorfer | Stark | 54.80 | Verified |
| 8 | Keokuk High School | A | Finch | Weilandich | Schilling | Joy | NT | Unverified: |