| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | East Bay |
A | Hall | Jackson | Sexton | Marshall | 52.48 | FHSAA 3A District 9 |
| 3 | Bloomingdale |
A | Exil | Hayes | Courtney | Falcon | 50.98 | Newsome Wolves Senior Night 2025 |
| 4 | Lennard |
A | Jorden | Sampson | Callaway | Reynoso | 47.61 | FHSAA 4A Region 3 |
| 5 | Riverview |
A | Fanchi | Lee-Shung | Campbell | Garlic | 48.32 | 2026 Nash Higgins Relays |
| 6 | Morgan High School | A | Payton | Carter | Brown | Henry | 51.25 | Morgan Inaugural Track Meet |
| 7 | Sumner |
A | Rankin | Brown | Barton | Jackson | 52.90 |