| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Ste. Genevieve High School | A | Amschler | Jokerst | Merriman | Lacey | 1:41.72 | Verified |
| 2 | Central (New Madrid County) High | A | Anderson | Henderson | Jones | Bennett | 1:36.80 | Verified |
| 3 | Dexter High School | A | Hinson-Flores | Farmer | McDonald | CUEVAS | 1:32.85 | Verified |
| 4 | Notre Dame (Cape Girardeau) High | A | Bruenderman | Eftink | Dow | Brown | 1:31.06 | Verified |
| 5 | Kennett High School | A | Boatman | Walls | Henson | Rodgers | 1:32.56 | Verified |
| 6 | Central (Park Hills) High School | A | Petty | Welch | Dalton | Bockenkamp | 1:34.44 | Verified |
| 7 | Kelly High School | A | Hileman | Appleton | Burleson | Birk | 1:38.11 | Verified |