| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | John Carroll |
A | Pittman | RICHARDSON | Coleman | Summerall | 44.66 | MCHS Sarge Invitational 2025 |
| 2 | Tradition Prep | A | Monaco | Frederic | Lopez | Richardson | 43.87 | Bulldog Invitational 2025 |
| 3 | Vero Beach |
B | Dagbolen | Burson | Lundy | Boatwright | 42.47 | FHSAA 4A District 8 |
| 4 | Centennial |
B | Rowley | Polynice | Harper | Johnson | 40.96 | Bill Wilson Invitational 2025 |
| 5 | Centennial |
A | Johns | Tolbert | Dorelus Jr. | Michel | 40.96 | Bill Wilson Invitational 2025 |
| 6 | Vero Beach |
A | Cabrera | Dillard | Hillsman | Jacobs | 42.47 | FHSAA 4A District 8 |
| 7 | Fort Pierce Westwood |
A | Ford | Lynch | Greer | Martin | 44.04 | Cobra Relays 2025 |
| 8 | Legacy High School | A | Rodriguez | Williams | Haynes | Serrano | NT |