| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Tradition Prep | A | Gay | Malvoisin | Mignott | Volcin | 45.86 | St. Lucie County Championships |
| 2 | Centennial | A | Sterling | Wright | Bacchas | Shabazz | 45.37 | FHSAA 4A District 10 |
| 3 | Treasure Coast | A | Clark | Reed | Davis | Harper | 43.98 | Cobra Relays 2023 |
| 4 | Fort Pierce Westwood | A | Wheeler | Vincent | Scucess | Mills | 43.54 | Cobra Relays 2023 |
| 5 | Fort Pierce Central | A | Tullis | Perceval | Gauthier | Haldane | 43.60 | Jupiter Relays 2023 |
| 6 | Port St. Lucie | A | Griffin III | Ngafeeson | Estinoble | Tolbert | DNS | Martin County Sarge Memorial Meet |
| 7 | John Carroll | A | Miller | Reynolds | Gibson | Alford | DNS | Holy Trinity Invitational |
| 8 | Lincoln Park | A | Berry | Grant | Platt | Thomas | NT |