| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Venice | A | Courville | Bartlett | Crettol | Lukowski | 9:41.31 | 2019 FHSAA State Track & Field Championships |
| 2 | Braden River | A | Marston | Auer | Davis | Busi | 10:15.81 | FHSAA 3A District 10 |
| 3 | Bishop Verot | A | Mills | Gulati | Thomas | Thomas | 10:16.05 | 2019 FHSAA State Track & Field Championships |
| 4 | Cardinal Mooney | A | Monserez | Jarrett | Smithers | Schaefer | 10:17.52 | FHSAA 1A District 7 |
| 5 | Charlotte | A | Winkler | Flowers | Piacitelli | Collins | 10:26.00 | |
| 6 | Sarasota | A | Menke | Belcher | Grimmer | Montidoro | 10:40.00 |