| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Hudson |
A | Gant | Foder | Kendrick | Doston | NT | |
| 3 | Central (Brooksville) |
A | Hall | Henry | Hill | Mitchell | 45.91 | 2026 Hernando Relays |
| 4 | Bishop McLaughlin |
A | Jester | Reynolds | Jacobs | Chalk | 46.71 | The Villages Home Meet Two |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Weeki Wachee |
A | Kernisant | Arellano | Wright | Abbinanti | 45.24 | FHSAA 3A District 8 |
| 3 | Wesley Chapel |
A | Walsh | Johnson | Lewis | Pineo | 41.69 | Pirate Booty Invitational- Approved Qualifier |
| 4 | Nature Coast |
A | Harvin | Warford | Montgomery | Mckenzie | 42.15 | FHSAA 2A Region 2 |
| 5 | Hernando |
A | Harris | Harris | Mason | Fagin | 45.91 | FHSAA 2A District 5 |