| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Innovation_Orlando | B | Moussa | Fitzherbert | Batts | Roderick | NT | |
| 4 | First Academy (Orlando) |
B | Herrington | Bird | Gardner | Robinson | NT |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Colonial |
A | Sukut | Smith | Rodriguez | Arroyo | NT | |
| 3 | Tohopekaliga |
A | Reid | Charles | Pagan | Francois | NT | |
| 4 | Lake Nona |
A | Rivera | Colon | Hamm | Vijayakumar | 2:02.46 | Poinciana Relays |
| 5 | Windermere |
A | Gil Rodriguez | Magnan | Rozhkova | Goote | DNS | |
| 6 | Innovation_Orlando | A | McNeill-Toro | Turner | Londoni | Behzad | NT | |
| 7 | First Academy (Orlando) |
A | Fones | Spafford | Schmid | Claytor | DQ | |
| 8 | University (Orlando) |
A | Acosta | Vassor | Gilmer | Ramirez | NT |