| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 1 | Pasco |
A | Lockridge | Garcia | Mares | Stinyard | 45.81 |
| 2 | Central (Brooksville) |
A | Henry | Hill | Mitchell | Mitchell | 45.35 |
| 3 | Weeki Wachee |
A | Wheeler | Ferrugia-Hughes | Wright | Abbinanti | 45.21 |
| 4 | Central (Brooksville) |
B | Espinoza | Henry | Hill | Mitchell | 45.35 |
| 5 | Hernando |
A | Harris | Harris | Brock | Fagin | 45.39 |
| 6 | Crystal River |
A | Johnson | Leal Hernandez | Wellington | George | 47.08 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 1 | Land O'Lakes |
A | Agbagu | Brown | Perez | Chalmers | 43.15 |
| 2 | Nature Coast |
A | Harvin | Warford | Montgomery | Mckenzie | 42.15 |
| 3 | Armwood |
A | Adams | Clark | Jones | Hudgins | 41.22 |
| 4 | Nature Coast |
B | Strong | Picariello | Edney | Woodard | 42.15 |
| 5 | Nature Coast |
C | Hannah | Speights | Cooper | Elwin | 42.15 |
| 6 | Weeki Wachee |
B | Kernisant | Kernisant | Maddox | Arellano | 45.21 |