| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Martin County | A | 53.28 | MCHS Tiger Invitational 2023 |
| 2 | St. Andrew's | A | 52.45 | Benjamin Meet #2 |
| 3 | Park Vista | A | 51.27 | FHSAA 4A Region 3 |
| 4 | Fort Pierce Central | A | 49.56 | Jupiter Relays 2023 |
| 5 | Dwyer | A | 49.86 | FHSAA 4A Region 3 |
| 6 | King's Academy | A | 51.31 | Dwyer Panther Relays |
| 7 | South Fork | A | DNS | FHSAA 3A Region 4 |
| 8 | Suncoast | A | 53.96 | Jupiter Warrior Invitational (17th Annual) |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Lake Worth | A | 59.00 | Kings Academy Invit 1 |
| 4 | Jupiter | A | 54.33 | Jupiter Relays 2023 |
| 5 | Forest Hill | A | 55.03 | FHSAA 4A District 11 |