| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 3 | Donahue Academy |
A | Fajardo | Iriarte | Shaw | Rook | 57.17 |
| 4 | First Baptist (Naples) |
A | Lugo | Pugh | Roth | Melancon | DQ |
| 5 | Bonita Springs |
A | Morales Carrion | Barragan | Kamel | Leonardo-Canales | 55.11 |
| 6 | Mason Classical Academy |
A | Mummert | Pierre Louis | Pierre-Louis | Rosario | 54.32 |