| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Brandon | A | 45.46 | 45.46 49th Nash Higgins Relays |
| 2 | Armwood | A | DNS | 44.65 Gaither Five Guys |
| 3 | Lennard | A | 44.26 | 44.26 Hillsborough County Championship |
| 4 | Tampa Bay Tech | A | 42.09 | 42.09 57th Annual Ed Wells Jr.-Kiwanis Invitational |
| 5 | Hillsborough | A | 42.88 | 42.88 Bob Hayes Invitational |
| 6 | King | A | 44.44 | 44.44 Wharton Wildcat Invite |
| 7 | Jesuit (Tampa) | A | 44.69 | 44.69 Jesuit Tri-Meet 2-27-19 |
| 8 | Leto | A | 46.44 | 46.44 Steinbrenner Quad-Meet 3-13 |