| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Glades Day |
A | Jones | Johnson | Williams | Villarreal | DNF | Pahokee Anquan Boldin Invitational 2025 |
| 4 | Pahokee |
A | Simms | Fitzgerald | Stinfort | Hill | 42.45 | FHSAA 1A Region 4 |
| 5 | Royal Palm Beach |
A | Rolle | Williams | Thomas | Curry | 43.63 | Royal Palm Beach Meet 2025 |