| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | SLAM (West Palm Beach) |
A | Sweeting | Pierre-Louis | Sosa | Carswell | NT | |
| 3 | South Fork |
A | Padgett | Holmes | Pepsidero | Churchill | 54.51 | Martin County Championships 2025 |
| 4 | Atlantic Christian |
A | Rollins | Hyppolite | Eugene | Moura | 53.60 | Clyde Harris Invitational at |
| 5 | Seminole Ridge |
A | Jean | Davies | Langsi | Allen | 54.23 | Dr. Garcia Invite #2 2025 |
| 6 | Jensen Beach |
A | Nguyen | Feurer | Carter | Laurent | 54.89 | Benjamin Invite #1 2025 (Sanctioned) |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Spanish River |
A | Ulysse | Theissen | Bethel | Fray | 51.85 | Dr. Garcia Invite #2 2025 |
| 2 | Santaluces |
A | Williams | Patillo | Casseus | Carra | 51.51 | PBC Meet 2025 |
| 3 | Treasure Coast | A | Delevery | Brown | Sama | Labbe | 50.51 | SLC Championships 2025 |
| 4 | Dwyer |
A | McClean | Gay | Odita | Smith | 49.35 | Cardinal Newman Invitational 2025 |
| 5 | Suncoast |
A | Brown | Johnson | Hester | Arp | 50.29 | Clyde Harris Invitational at |
| 6 | Inlet Grove |
A | Sharpe | Steward | Harper | Leveille | 51.48 | PBC Meet 2025 |
| 7 | Park Vista |
A | Haldas | Hernandez | Dawkins | Collins | 51.80 | Cardinal Newman Invitational 2025 |
| 8 | Dreyfoos |
A | Pierre | Cannon | Mcdow | Pierre | 53.54 | Royal Palm Beach Meet 2025 |