| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Lake Placid |
A | Charite | Bullard, Jr. | Walton | Smith | 4:15.93 | Lake Placid Meet #1 |
| 3 | DeSoto County |
A | Sandate | Diaz | Tice | Wilkerson | 3:55.02 | Sebring Relays 2024 |
| 4 | Sebring |
A | Albritton | Cruz | Hamilton | Volvaire | 3:39.03 | Sebring Relays 2024 |
| 5 | LaBelle |
A | Zander | Davis | Thompson | Blackstock | 3:52.51 | TFRRS 03/08/24 - B team No time |
| 6 | Avon Park |
A | jones | Roman | Silva | Langston | 4:02.72 | Lake Placid Meet #1 |
| 7 | Okeechobee |
A | Campbell | Stewart | Walsh | Williams | NT |