| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Bartow | A | 3:58.96 | 3:58.96 Davenport invitational 2023 (3/25/2023) |
| 4 | Poinciana | A | 3:42.14 | 3:42.14 Poinciana Relays (3/02/2023) |
| 5 | Haines City | A | 3:50.88 | 3:50.88 Lake Region - Thunder Relays2023 (3/28/2023) |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Winter Haven | A | 3:32.98 | 3:32.98 Kowboy Invitational (3/10/2023) |
| 3 | Plant City | A | 3:32.49 | 3:32.49 2023 Hillsborough County Championships (4/12/2023) |
| 4 | George Jenkins | A | 3:22.41 | 3:22.41 Berkley Track and Field Invitational (3/04/2023) |
| 5 | Ridge Community | A | 3:31.60 | 3:31.60 Polk County Championship 2023 (4/08/2023) |
| 6 | Celebration | A | 3:32.54 | 3:32.54 Eye of the Storm Invitational (4/07/2023) |
| 7 | Lake Gibson | A | 3:41.89 | 3:41.89 Lakeland City meet @ George Jenkins 2023 (3/30/2023) |