| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Discovery |
A | Cariotte | Green | Forgues | Sewnarine | 4:17.19 | 4:17.19 Polk County Championship 2023 |
| 4 | Frostproof |
A | Griffiths | Gammage | Pittman | Ramirez | 3:50.51 | FHSAA 2A Region 3 |
| 5 | Bartow |
A | Brinson | Valentine | Robinson | Robinson | 3:58.96 | 3:58.96 Davenport invitational 2023 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Tenoroc |
A | Davis | Garrett | Guerrero | Thomas | 3:47.32 | Winter Haven Classic |
| 2 | Davenport |
A | George | Thomas | Jean-Jacques | Diaz DeJesus | 3:35.63 | 3:35.63 Poinciana Relays |
| 3 | Dr. Phillips |
A | Hardy | Butler | Scott | Daniels | 3:30.00 | St. Cloud Invitational |
| 4 | Lake Wales |
A | Daniels | Davis | Filsaime | Jean | DNS | |
| 5 | Auburndale |
A | Johnson | Cuyler | Jiles | Russell | 3:27.68 | 3:27.68 Lake Region - Thunder Relays2023 |
| 6 | Cocoa |
A | Bass | Glover | Simmons-Roberts | Smith | 3:32.54 | FHSAA Class 2A Track & Field State Championships |
| 7 | Kathleen |
A | Chappell | Abram | Wilson | Brito | DNS | 3:45.99 Lakeland City meet @ George Jenkins 2023 |
| 8 | East Ridge |
A | Moore | jordan | Rivers | Smith | 3:50.05 | 3:50.05 Trinity Prep Relays 2023 |