| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Jupiter Christian |
A | 53.15 | Calvary Christian FTL Varsity Meet #1 |
| 4 | Fort Pierce Central |
A | 51.80 | Cobra Relays 2025 |
| 5 | A | 52.13 | Dr. Garcia Meet #1 |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Jupiter |
A | 51.30 | Benjamin Invite #2 2025 (Sanctioned) |
| 3 | Treasure Coast | A | 50.11 | The Lincoln Knowles Invitational 2025 |
| 4 | Dwyer |
A | 48.83 | FHSAA 4A Region 2 |
| 5 | Suncoast |
A | 49.09 | FHSAA 3A Region 4 |
| 6 | Martin County |
A | 50.18 | FHSAA 4A Region 2 |
| 7 | Dreyfoos |
A | 51.61 | Royal Palm Beach Invitational 2026 |