| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Jensen Beach |
A | Cewe | Chrysler | Sullivan | Huston | 46.00 | |
| 3 | Martin County |
A | Oh | Allison | Ozuna | Ocampo | 44.96 | Cobra Relays 2025 |
| 4 | Centennial |
A | Benito | Thomas | Remeus | Raphael | DNS | Bill Wilson Invitational 2025 |
| 5 | Fort Pierce Central |
A | Blackmon | Sanon | Daughtry | Brown | 43.75 | Cobra Relays 2025 |
| 6 | South Fork |
A | Bailey | Miguel | Rodriguez | Valenti | 45.12 | Jensen Beach Invitational 2026 |
| 7 | Lincoln Park |
A | Davis | Rene | Cunningham | Goldberg | 46.81 | SLC Championships 2025 |