| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Hastings | A | Osgood | Schaaf | Buresh | Chavez Gutierrez | NT | |
| 3 | Holdrege | A | Michalski | Rooks | Hale | Vaughn | 1:01.00 | |
| 4 | Minden | A | Hansen | Petersen | Gramke | Klabunde | 1:00.00 | |
| 5 | Lexington | A | Wolf | Carlson | Sutton | Ortiz-Ibarra | 1:00.00 | |
| 6 | Pleasanton | A | Mata | Hawkins | Stark | Hawkins | 1:08.56 |