| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | South Florida HEAT | A | Waters | Godbey | Christensen | Godbey | 51.99 | Saint Andrew's HS Track Invitational #1 |
| 4 | Jensen Beach | A | Washington | Chrysler | Hall | Ahern | 45.57 | South Fork Meet #2 |
| 5 | Lincoln Park | A | Platt | Berry | Grant | Richardson | 45.89 | St. Lucie County Meet 2023 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Dwyer | A | Elie | Shrank | Gilbert | Key | 45.02 | Panther Relays 2023 |
| 2 | Port St. Lucie | A | Tolbert | Estinoble | Ngafeeson | Griffin III | DNF | Martin County Sarge Memorial Meet |
| 3 | Fort Pierce Central | A | Pierre-louis | Parks | Kennedy | Stinson | 43.60 | Jupiter Relays 2023 |
| 4 | Martin County | A | Hochstein | Truitt | Smith | Alexander | 42.38 | FHSAA Class 4A & Para Track & Field State Championships |
| 5 | Fort Pierce Westwood | A | Vincent | Wheeler | celisca | Mills | 43.54 | Cobra Relays 2023 |
| 6 | Sebastian River | A | Taylor | Campbell | McKenzie | Grimsley | 43.84 | FHSAA 3A District 13 |
| 7 | South Fork | A | Borgelin | Sammons | St. Louis | Mussenden | 44.98 | FHSAA 3A District 14 |
| 8 | Centennial | A | Boone | Sterling | Bacchas | Jean-Baptiste | 45.04 | St. Lucie County Meet 2023 |