| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Marion County High School | A | Gerding | Kindhart | Walker | Whetstone | 1:05.00 | Unverified: |
| 2 | Scotland County High School | A | NT | Unverified: | ||||
| 3 | South Shelby High School | A | Bowen | Lanpher | Elsen | Elliott | 48.97 | Verified |
| 4 | Highland High School | A | Stark | Harmon | Dare | Ritterbusch | 46.08 | Verified |
| 5 | Brookfield High School | A | Howerton | Mitch | Lumsden | Fields | 48.26 | Verified |
| 6 | Palmyra High School | A | Durst | Smith | Kroeger | Juarez | 50.58 | Verified |
| 7 | Keokuk High School | A | Schilling | Hays | Williams | Davis | 1:00.00 | Unverified: |
| 8 | Clark County High School | A | NT |
