| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 2 | Parrish | A | Hough | Gordillo | Vitoritt | Scanlan | NT |
| 3 | Southeast | A | Thomas | Davis | Perkins | Williams | 44.88 |
| 4 | Braden River | A | Hogan | Thomas | Stephens | Kelly | 42.24 |
| 5 | Palmetto | A | Dortch | Hughes | Dawes | Stamp | 42.70 |
| 6 | Manatee | A | Sanders | Jackson | Bean | Jackson | 45.50 |