| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Cypress Creek (Wesley Chapel) |
A | Palmer | Murray | Carrero | Wells | 10:23.50 | George Steinbrenner Invitational (Approved Qualifier) |
| 2 | Sunlake |
A | Pope | Millen | Pereira | Ford | 10:24.95 | East Lake Early Bird Invitational (Approved Qualifier) |
| 3 | Land O'Lakes |
A | Lopez | Bullough | Sanderson | Homer | 11:01.78 | Tampa Jesuit Meet 5 |
| 4 | Wiregrass Ranch |
A | Gregory | Burkett | Powers | Spearin | 11:02.24 | Coyote Invitational |
| 5 | Wesley Chapel |
A | Michel | Gattuso | Walls | Johnson | 11:07.26 | Raven Invitational 2026 (FHSAA Sanctioned) |
| 6 | Zephyrhills |
A | Audet | Farah | Williams | Kane | NT |