| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | First Baptist (Bunnell) | A | Jarrett | Young | Starrett | Arneaud | NT | |
| 3 | St. Joseph | A | Vilorio | Casey | Doctora | Montgomery | 56.00 | |
| 4 | Matanzas | A | Wright | Miller | Dayton | Gaines | 52.34 | George Janan SC Invitational M |
| 5 | Crescent City | A | Green | Onofre | lopez | Joely | 55.81 | FHSAA Class 2A-District 6 |
| 6 | Matanzas | B | Zepponi | Giles | Zysek | Marcellus | 59.00 |