| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Cardinal Newman |
A | Jimenez | Monegro | Suarez-de-puga | Nunez | NT | |
| 4 | Seminole Ridge |
A | Prince | Coffey | Langsi | Jean | 55.08 | Royal Palm Beach Meet 2025 |
| 5 | SLAM (Boca Raton) |
A | Kuehne | Nieves | Vasquez | Calabrese | 1:01.63 | St Andrews Varsity |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Park Vista |
A | Fleuricin | Haldas | Brown | Collins | 52.36 | Panther Relays 2025 |
| 3 | Lake Worth |
A | Clayton | Jones | Greenwich | Razz | 51.96 | Jeff Fossen Invitational 2025 |
| 4 | Dwyer |
A | McClean | Gay | Jean | Odita | 49.80 | Bob Hayes High School Invitational |
| 5 | Palm Beach Gardens |
A | Mcgee | Perry | Collier | Hooks | 50.12 | PBGHS Invitational 2025 |
| 6 | West Boca Raton |
A | Mignano | Whitfield | Julien-Jeune | Gonzales | 51.99 | Park Vista #1 2025 |
| 7 | Boca Raton |
A | Aguilar | Dobson | Saint-Louis | Faustin | DNS | Jeff Fossen Invitational 2025 |