| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | West Oaks |
A | Laventure | Ards | Ingram | Cole | DNF | Larry Kelly Invitational 2026 |
| 2 | South Lake |
A | Powell | Swaby | McKee | White | DNS | Evans Development Invitational |
| 3 | Edgewater |
A | Pipkin | Aicher | Cleveland | Sterling | 43.13 | Bishop Moore - Carl Friedland 2026 |
| 4 | Evans |
A | Hickson | Hardy | Mincey | Jacobs | 42.03 | Florida Relays |
| 5 | Jones |
A | Johnson | Brown | Bell | McCluster | 42.08 | 2026 Dr. Baptiste Invitational hosted by West Orange High |
| 6 | Boone |
A | Noelsaint | Allen | Hardy | Woods | 43.24 | Bishop Moore - Carl Friedland 2026 |
| 7 | Ocoee |
A | Cidel | Lipscomb | Holley | Paul | DNF |