| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Donahue Academy |
A | Lebrun | Mariano | Jaramillo | Rankin | 3:59.82 | Cypress Panther Invitational |
| 4 | Village School |
A | Leadbetter | Germain | Stepas | Sporn | 3:36.65 | FHSAA 1A District 12 |
| 5 | Mason Classical Academy |
A | Alce | Conde | Finn | Grajales-Franco | 3:46.35 | 62nd Eagle Invitational |
| 6 | Marco Island Academy |
A | Brinker | Juels | Uristegui | Keim | NT |