| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Seminole Ridge |
A | Allen | Alvin | H | Langsi | 4:41.60 | John I Leonard Invitational |
| 3 | Palm Beach Central |
A | Echavarria | Anderson | Ter-Avanesova | Tortora | 4:36.84 | Dr Garcia Invite #1 2025 |
| 4 | Park Vista |
A | Bell | Joly | Chu | Massena | 4:03.17 | Panther Relays 2025 |
| 5 | Centennial |
A | Schmook | Robinson | Williams | Hoskins | 4:12.02 | Jupiter Warrior Invitational 2025 |
| 6 | Oxbridge Academy |
A | Duke | Wajer | Martin | Wangombe | 4:41.34 | John I Leonard Invitational |
| 7 | Dr. Joaquin Garcia | A | Paciletti | Gring | Murphy | Brown | 4:45.33 | Panther Relays 2025 |