| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Spoto |
A | 8:14.48 | FHSAA 3A Region 3 |
| 2 | Bloomingdale |
A | 8:18.59 | FHSAA Class 4A Track & Field State Championships |
| 3 | Hillsborough |
A | 8:20.00 | |
| 4 | Alonso |
A | 8:28.18 | FHSAA 4A District 6 |
| 5 | Palm Harbor University |
A | 8:29.09 | Pinellas County Athletic Conference (PCAC) |
| 6 | Plant City |
A | 8:45.00 | |
| 7 | Robinson |
A | 8:52.65 | FHSAA 2A Region 3 |
| 8 | Armwood |
A | 9:00.20 | 9:00.20 Hernando Kiwanis Invite |
| 9 | Leto |
A | 9:04.96 | 9:04.96 2023 Manuel-Griffin-Fraundorfer Relays |
| 10 | Middleton |
A | 9:09.94 | 9:09.94 WC National Division Meet |
| 11 | Wiregrass Ranch |
A | 9:11.11 | |
| 12 | Lennard |
A | 9:15.00 | |
| 13 | Riverview |
A | 9:16.15 | 9:16.15 WC Federal Division |
| 14 | Tampa Bay Tech |
A | 9:21.00 | |
| 15 | Winthrop College Prep | A | 10:13.19 | IMG Academy Invitational |
| 16 | Blake |
A | NT | |
| 17 | Chamberlain |
A | NT | |
| 18 | Durant |
A | NT | |
| 19 | Newsome |
A | NT | |
| 20 | Strawberry Crest |
A | NT |