| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Heritage |
A | Clarks | Ferguson | Jean-Louis | Melendez | NT | |
| 3 | St. Edward's |
A | Callaghan | Ramsay | Hebig | Storts | 5:18.39 | Vero Beach HS 2026 Preseason Classic |
| 4 | Vero Beach |
A | Domashinski | Wells | Gibson | Ewar | 4:00.54 | St. Cloud Invitational (Qualifier) |
| 5 | John Carroll |
A | Martinez | Delince | VAN RENS | VILLANUEVA | 4:24.40 | Cobra Relays 2026 |
| 6 | Lincoln Park |
A | Brown | Daniel | Anderson | Whilby | NT | |
| 7 | Fort Pierce Central |
A | Veal | Tommie | Andrews | Wilder | NT |