| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Jensen Beach |
A | Nguyen | Laurent | Feurer | Carter | DQ | Benjamin Invite #1 2025 (Sanctioned) |
| 3 | Martin County |
A | Ebanks | Edwards | Garcia | Perez | 52.62 | Cobra Relays 2025 |
| 4 | Fort Pierce Westwood |
A | Villegas | Brown | Angervil | Figaro | 49.45 | Bill Wilson Invitational 2025 |
| 5 | Somerset College Prep | A | Bourne-Morgan | Lewis | Miller | Wright | 51.88 | SLC Championships 2025 |
| 6 | Tradition Prep | A | Parrel | Derius | Falcon | Chavers | 54.79 | Cobra Relays 2025 |
| 7 | Jupiter Christian |
A | Schiralli | Zawadski | Winslow | Winslow | 55.31 | WA Relays |