| Lane | Athlete | Team | Seed | 
|---|---|---|---|
| 2 | CARROLL, Christian | Bartram Trail | 44.50 | 
| 3 | LANGELAND, Connor | Bartram Trail | 42.13 | 
| 4 | GOMBAR, Caleb | Bartram Trail | 41.06 | 
| 5 | MONTERO, Enrico | Fleming Island | 41.72 | 
| 6 | HAMMETT, Jaycob | Bartram Trail | 42.80 | 
| 7 | SAVAGE, Avery | Fleming Island | NT |