| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Tradition Prep | A | Chavers | Falcon | Derius | Parrel | 54.79 | Cobra Relays 2025 |
| 2 | Fort Pierce Central |
A | Simeus | Bolden | Lane | Stinson | 51.80 | Cobra Relays 2025 |
| 3 | Centennial |
A | Hoskins | Schmook | Schmook | Williams | 51.01 | Dr. Garcia Invite #2 2025 |
| 4 | Fort Pierce Westwood |
A | Brown | Figaro | Holmes | Villegas | 49.45 | Bill Wilson Invitational 2025 |
| 5 | Lincoln Park |
A | Bacon | Huskin | Lundy | Brown | 50.95 | Bill Wilson Invitational 2025 |
| 6 | Treasure Coast | A | Smith | Brown | Sama | Labbe | 51.30 | Jupiter Warrior Invitational 2025 |
| 7 | Somerset College Prep | A | Bourne-Morgan | Wright | Miller | Santiago | 52.62 | PBGHS Invitational 2025 |