| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 4 | Kathleen |
A | Brito | Brito | Bankston | Kelly | DNS | |
| 5 | Lake Wales |
A | Alls | Jean | Daniels | Filsaime | NT |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Tenoroc |
A | Garrett | Furman | Delu | Davis | 49.87 | Poinciana Relays |
| 2 | Cocoa |
A | Chappel | Chappel | Chassion | Smith | 44.00 | |
| 3 | Dr. Phillips |
A | Scott | Hardy | Fortilien | Horne | 42.95 | Sam Burley Hall of Fame Invitational |
| 4 | Bartow |
A | Murray | Brown | Williams | Taylor | 42.85 | |
| 5 | Auburndale |
A | Underwood | Russell | Jiles | Cuyler | 42.91 | FHSAA Class 3A Track & Field State Championships |
| 6 | Davenport |
A | Jean-Jacques | Chin-suer | Peters | Raiford | 43.83 | 43.83 Poinciana Relays |
| 7 | Frostproof |
A | Daniels | Plain | Blocker | Solomon | 45.35 | FHSAA 2A Region 3 |
| 8 | Discovery |
A | DNS | Poinciana Relays |