| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | DeLand |
A | DNS | SCHS Developmental Meet 1 February 18 2026 |
| 2 | Lake Mary |
A | 4:20.68 | Hagerty Track and Field Invitational 2026 |
| 3 | University (Orlando) |
A | 4:13.24 | Hagerty Track and Field Invitational 2026 |
| 4 | Wekiva |
A | DNS | FHSAA 4A Region 1 |
| 5 | Seminole (Sanford) |
A | 3:56.83 | FHSAA 4A Region 1 |
| 6 | Matanzas |
A | 4:17.41 | Lake Mary Invitational 2025 |
| 7 | Lyman |
A | 4:42.80 | SAC Freshman Sophomore Championship 2025 |
| 8 | Father Lopez |
A | NT |