| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | John Carroll | A | Pittman | Singleton | Gibson | Reynolds | DNF | Holy Trinity Invitational |
| 4 | Centennial | A | Sterling | Riveron | Shabazz | Bacchas | 45.37 | FHSAA 4A District 10 |
| 5 | Sebastian River | A | Godwin | Cummings | McKenzie | Campbell | 45.64 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Treasure Coast | A | Davis | Clark | Issac | Harper | 44.04 | FHSAA 4A District 10 |
| 3 | Fort Pierce Central | A | Tullis | Thompson | Beauford | Perceval | 43.60 | Jupiter Relays 2023 |
| 4 | Martin County | A | Lewis | Smith | Smith | Alexander | 42.38 | FHSAA Class 4A & Para Track & Field State Championships |
| 5 | Tradition Prep | A | Marshall | Delphin | Russell | Volcin | 43.00 | |
| 6 | Port St. Lucie | A | Ellis | Felder | Tolbert | Griffin III | 43.99 | Martin County Sarge Memorial Meet |
| 7 | Fort Pierce Westwood | A | Wheeler | Mills | Scucess | Vincent | 44.21 | Spanish River Invitational 2023 |