| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Osceola (Seminole) | A | BERRIAN | LE | Khammanivong | HARROLD | 45.80 | |
| 2 | Pinellas Park | A | Adams | Kier | Hampton III | Holston Jr. | DNS | |
| 3 | Boca Ciega | A | Jones | Merriex | Allen | Jackson | DNS | FHSAA 3A Region 3 |
| 4 | Lakewood | A | Mack | John | Adams | Morgan | 42.33 | Richard Allen Invitational |
| 5 | Palmetto | A | Stamp | Davis | Dawes | Hughes | 42.80 | |
| 6 | Largo | A | Bell | Randall | Kier | Williams | 43.27 | Seminole Warhawk Invitational |
| 7 | Seminole | A | Hartog | Christopher | Peterson | Marr-Navas | DNS | 45.78 Warhawk Invitational |