| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Somerset College Prep | A | Alperis | Elipha | Hill | Gurrier | 46.78 | Pahokee Anquan Boldin Invitational 2025 |
| 2 | John Carroll |
A | Pittman | Smith | Johnson | Housing | DNS | Benjamin Invite #1 2025 (Sanctioned) |
| 3 | Treasure Coast | A | Harper | Estassin | Jones | Richardson | 44.80 | Cobra Relays 2025 |
| 4 | Centennial |
A | Johns | Dorelus Jr. | Polynice | Jean-Pierre | 40.96 | Bill Wilson Invitational 2025 |
| 5 | Tradition Prep | A | Richardson | Atwell | Lopez | Drummond | 43.87 | Bulldog Invitational 2025 |
| 6 | Fort Pierce Westwood |
A | Boone | Ford | Lynch | Mckenzie | 44.91 | TFRRS 03/07/25 - 43.89 |
| 7 | Fort Pierce Central |
A | Hills | Reynolds | Saint-Jean | Ransfer | 46.52 | Palm Bay Invitational |
| 8 | Lincoln Park |
A | Chagani | Holland | Eshleman | Kapsis | 47.11 | Cobra Relays 2025 |