| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Indian Trails | A | 5:23.69 | East Coast Fr/So & MS Invitational |
| 3 | St. Johns Country Day | A | 5:00.02 | 5:00.02 Clay County Championships |
| 4 | Episcopal | A | 3:56.96 | FHSAA Class 2A/Para-Athlete Track & Field State Championship |
| 5 | Assumption Catholic School | A | 4:45.28 | |
| 6 | Interlachen | A | 5:15.00 | |
| 7 | St. Elizabeth Ann Seton Knight | A | 5:35.00 |