| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | St. Edward's |
A | Bryant | Cannon | Perez | Neumann | 55.04 | FPC Fresh/Soph Meet 2025 |
| 2 | Centennial |
A | Green | Cage | Schmook | Hoskins | 49.69 | FHSAA 4A Region 2 |
| 3 | Fort Pierce Westwood |
A | Blaise | Villegas | Brown | Jackson | 49.45 | Bill Wilson Invitational 2025 |
| 4 | Vero Beach |
A | Messam | Lundy | Moise | Wells | 47.77 | Bill Wilson Invitational 2025 |
| 5 | Vero Beach |
B | Taylor | Maldonado | Tomlin | Smith | 47.77 | Bill Wilson Invitational 2025 |
| 6 | Centennial |
B | Chery | Sinclair | Schmook | Miller | 49.69 | FHSAA 4A Region 2 |
| 7 | Tradition Prep | A | Parrel | Falcon | Derius | Phillippe | 54.22 | Atlantic HS Last Chance |
| 8 | St_Andrews_Episcopal |
A | Roccasecca | Courtemanche | Keel | Noel | DNS |