| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Montverde Academy |
A | Critoph | Shaban | Verma | Truong | DNS | |
| 4 | Pineapple Cove |
A | Jones | Rutledge | Murphy | Parker | DNS | |
| 5 | Cornerstone Charter |
A | Cumbal | Daly | Patrizio | Reynoso | 4:04.97 | Pioneer Invitational |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Hagerty |
A | Gandy | Henson | Rhodes | Shyy | DNS | |
| 3 | Foundation Academy |
A | Cannon | Nyland | McFarland | Jhonson | 3:31.48 | FHSAA 1A Region 2 |
| 4 | Edgewater |
A | Meadows | Wilkerson | Cleveland | Auguste | 3:21.72 | |
| 5 | IMG Academy |
A | Michelin | Kalamaras | McKenzie | Falk | DNS | |
| 6 | Victory Christian (LL) |
A | Cunningham | Paul | Williams | Morrison | 3:34.00 | |
| 7 | First Academy (Orlando) |
A | Crouch | Hughes | Ivsic | Wilson | DNS | FHSAA 1A Region 2 |