| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | LaBelle |
A | Williams | Jackson | Wheatle | Sandoval | NT | |
| 3 | Moore Haven |
A | Rembert | Santana | Walker | Perez | 1:54.10 | Sebring Relay Meet 2025 |
| 4 | Okeechobee |
A | Hebel-Hoover | Nesby | Barrett | Rochelle | 1:56.80 | Sebring Relay Meet 2025 |