| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Columbia West MS | A | Russel | Adams | Brinkley | Peterson | 1:00.40 | Verified |
| 3 | Columbia Smithton MS | B | Shen | Davis | Monnig | Coffman | 59.99 | Verified |
| 4 | Columbia Smithton MS | A | Adam | Eversole | Watson | Gibson | 59.99 | Verified |
| 5 | Columbia West MS | B | Pipes | Harper | Popp | Kateti | 1:00.40 | Verified |
