| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Frontier Middle School | A | Henriquez | Kelley | McDonnell | Weishaar | 1:06.15 | Verified |
| 2 | Wright City MS | A | Kirn | Mikus | Rees | Clubb | 59.59 | Verified |
| 3 | Troy South MS | A | Getz | Meyers | Schulte | Hopkins | 58.74 | Verified |
| 4 | Hannibal Middle School | A | Reese | Mayfield | Karr | Perry | 56.77 | Verified |
| 5 | Black Hawk Middle School | A | Daniel | Kerpash | Vacek | Morris | 58.27 | Verified |
| 6 | Wentzville South Middle School | A | Lappe | Schultz | Wall | Pender | 58.78 | Unverified: Wentzville Tri Meet |
| 7 | Troy Middle School | A | Colbert | Dunard | Bachmann | Croxton | 1:00.39 | Verified |
| 8 | Troy South MS | B | NT |
