| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Jensen Beach |
A | Bradley | Laurent | Trimble | Peruski | DNS | Jensen Beach Invitational 2026 |
| 3 | West Boca Raton |
A | Whitfield | Otero | Joseph | Julien-Jeune | 51.35 | Palm Beach Gardens Invite #1 2026 |
| 4 | Fort Pierce Central |
A | Alexander | Fequiere | Stinson | Simeus | 51.21 | Cobra Relays 2026 |
| 5 | Martin County |
A | Fernandes | Perez | Osdon | Ebanks | 51.28 | Jensen Beach Invitational 2026 |
| 6 | Glades Day |
A | Ocanas | Baez | Sturgis | Sisnett | 54.10 | Glades Central Invite 2026 |
| 7 | Pine School |
A | DeGeorge | Kaliniak | Hildebrand | Klostreich | NT |