| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 1 | Weeki Wachee |
A | Hessler | Anderson | Williams | Randazzo | NT |
| 2 | Dunnellon |
A | Hatcher | De La Rosa | Fenton | Alvord | 4:49.13 |
| 3 | South Sumter |
A | Cruz | Davis | Rodgers | Guevarra | 4:34.12 |
| 4 | Armwood |
A | Thornton | Ankers | Allen | Archie | 4:36.92 |
| 5 | Nature Coast |
A | Dardignac | Anderson | Sanders | Smith | 4:51.53 |
| 6 | Crystal River |
A | Broadbent | Cordero-Santana | Johnson | Santiago | 5:04.78 |