| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Smith-Cotton High School | A | Powell | Taylor | Mackin | Bell | 53.14 | Verified |
| 2 | Rock Bridge High School | A | Lyson | Miller | Gerrish | Fedenia | 51.15 | Verified |
| 3 | Capital City High School | A | Strayhorn | Sterrett | Cris-Kargbo | Hess | 50.17 | Verified |
| 4 | Helias Catholic High School | A | Phillips | Crisostomo | Harrison | Struemph | 49.51 | Verified |
| 5 | Battle High School | A | Weitkemper | Brookins | Johnson | Jones | 49.81 | Verified |
| 6 | Hickman High School | A | Briscoe | Hoffman | Smith | Maland | 50.82 | Verified |
| 7 | Jefferson City High School | A | Okpodighe | Graham | Houston | Hurtalt | DQ | Verified |