| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Lake Mary Prep | A | DNS | Verified |
| 2 | Bishop Moore HS | A | DNF | Verified |
| 3 | Winter Park HS | A | 49.71 | Verified |
| 4 | Sanford Seminole HS | A | 46.64 | Verified |
| 5 | Colonial HS | A | DQ | Verified |
| 6 | Hagerty HS | A | 52.20 | Verified |
| 7 | Foundation Academy | A | 1:02.05 | Verified |
| 8 | Winter Springs HS | A | DNS | Unverified: |