| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | George Steinbrenner | A | 51.02 | Steinbrenner Tri-Meet 3-6-19 |
| 3 | Alonso | A | 50.56 | Kowboy Invitational 2019 |
| 4 | Tampa Bay Tech | A | 49.55 | Wharton Wildcat Invite |
| 5 | Lakeland | A | DNF | Lakeland City Meet 2019 |
| 6 | George Jenkins | A | DQ | Charger Invitational |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 3 | Sunlake | A | 51.72 | Mitchell Meet #2 |
| 4 | Wiregrass Ranch | A | 51.36 | Wharton Wildcat Invite |
| 5 | Land O'Lakes | A | 51.56 | FHSAA 3A Region 2 4/26/18 |
| 6 | Palm Harbor University | A | 52.17 | Warhawk Invitational |
| 7 | Mitchell | A | 59.36 | 57th Annual Ed Wells Jr.-Kiwanis Invitational |