| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | George Steinbrenner |
A | 45.72 | Freedom Spring Break Invitational |
| 3 | Hillsborough |
A | 44.81 | 2026 Nash Higgins Relays |
| 4 | Sickles |
A | 44.17 | Tampa Jesuit Meet 3 |
| 5 | Cypress Creek (Wesley Chapel) |
A | 44.71 | Freedom Spring Break Invitational |
| 6 | Robinson |
A | 44.93 | Freedom Spring Break Invitational |
| 7 | Angeline Academy | A | 47.52 | West Coast Invite |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | King |
A | 43.84 | Newsome Wolves Invitational 2026 (Approved Qualifying Meet) |
| 2 | River Ridge |
A | 43.68 | East Pasco Invitational |
| 3 | Wiregrass Ranch |
A | 42.54 | Coyote Invitational |
| 4 | Wesley Chapel |
A | 41.69 | Pirate Booty Invitational- Approved Qualifier |
| 5 | Wharton |
A | 42.05 | 2026 Manuel Griffin Fraundorfer Relays |
| 6 | Newsome |
A | DNS | 2026 Manuel Griffin Fraundorfer Relays |
| 7 | Alonso |
A | 43.71 | 2026 Manuel Griffin Fraundorfer Relays |
| 8 | Sunlake |
A | 43.91 | West Coast Invite |