| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Lake Highland Prep | A | Kenoyer | Walton | Phillips | Miller | 8:05.26 | 2019 FHSAA State Track & Field Championships |
| 2 | Timber Creek | A | Barron | Isham | Evans | Richardson | 8:16.70 | FHSAA 4A District 4 |
| 3 | Lake Mary | A | Arauma | Farrell | Spurling | Miranda | 8:19.26 | FHSAA 4A Region 1 and FHSAA 2A Region 2 |
| 4 | Lyman | A | Vaughn | Kuzmick | Hodges | Shah | 8:26.58 | FHSAA 4A District 2 |
| 5 | Colonial | A | Giron | Rojas | Velez | Rosado | 8:30.46 | FHSAA 4A District 4 |
| 6 | East Ridge | A | Kaden | Guevara | Lugo Ramos | Melendez | 8:31.82 | Bob English Relays 2020 |
| 7 | Geneva School | A | White | Mckay | Shafer | Gibson | 8:59.39 | FHSAA 1A District 9 |
| 8 | Lake Minneola | A | Bradshaw | Bradshaw | Rojas | Reed | 9:00.00 | |
| 9 | West Oaks | A | Cheristin | Alcius | Leverett | McKinnis | NT |