| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Okeechobee | A | Johns-Chilsom | Marables | Bryant | Williams | 43.96 | FHSAA 3A Region 4 |
| 3 | Fort Pierce Central | A | Kennedy | Reynolds | Dieujuste | Stinson | 43.60 | Jupiter Relays 2023 |
| 4 | Cardinal Newman | A | Redmon | Miller | Desouza | Levy | DNS | FHSAA Class 1A Track & Field State Championships |
| 5 | Martin County | A | Lewis | Smith | Smith | Alexander | 42.38 | FHSAA Class 4A & Para Track & Field State Championships |
| 6 | Bayside | A | Josephs | Craig | Parkes | Reynolds | 43.82 | Merritt Island Open |
| 7 | Port St. Lucie | A | Ngafeeson | Tolbert | Estinoble | Griffin III | 43.99 | Martin County Sarge Memorial Meet |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Jensen Beach | A | Washington | Palmer | 47.01 | MCHS Tiger Invitational 2023 | ||
| 4 | South Fork | A | Borgelin | Mussenden | St. Louis | Sammons | DNS | FHSAA 3A District 14 |
| 5 | Centennial | A | Bacchas | Boone | Wright | Sterling | 45.37 | FHSAA 4A District 10 |