| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Treasure Coast Classical | A | Wojcieszak | Griffin | Hook | Smith | NT | |
| 4 | Centennial |
A | Hoskins | Elma | Mitton | Green | 4:20.48 | 28th Annual Warrior Invitational 2024 |
| 5 | Martin County |
A | Anderson | Cantu | Lindsay | Rapaport | 4:24.95 | Falcon Classic 2024 |
| 6 | Fort Pierce Central |
A | Barrios | Fortin | Roper | Wilder | NT |