| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Timber Creek | A | D'Allesandro | Varnedore | Varnedore | Sico | 8:31.00 | St. Cloud Invite |
| 2 | Freedom (Orlando) | A | Lespire | Pardo | Persaud | Falcon | 8:34.48 | FHSAA 4A Region 1 and FHSAA 2A Region 2 |
| 3 | Lake Nona | A | Nunn | Pack | Barker | Grace | 8:44.22 | Nona |
| 4 | East River | A | Casey | Folse | Hanlin | Hickman | 8:45.00 | |
| 5 | Winter Park | A | Fuller | Patil | Creesey | Leichner | 9:01.84 | Toho Invite 2020 |
| 6 | University (Orlando) | A | Legzouli | Blandino | Brockhoff | Shields | 9:20.00 | |
| 7 | Colonial | A | Marrero | Reyes | Rojas | Velez | 9:25.12 | Practice |
| 8 | Cypress Creek | A | Lewis | Gonzalez | Torres Plaza | Feliz | 9:30.45 | Cypress Creek |
| 9 | Boone | A | Charles | Correa | Shea | Petersen | 10:15.00 | Boone |
| 10 | Oak Ridge | A | SILVERN | JACKSON | SAINT MARC | PEREZ | 11:56.23 | ORHS |