| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Manatee |
A | Woods | Smalls | Dorce | Moody | NT | |
| 3 | Cardinal Mooney |
A | Wren | Borda | Mearriam | Koch | 51.84 | FHSAA 2A District 11 |
| 4 | Lakewood Ranch |
A | Harris | Abreu | McBean | Frint | 53.00 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | North Port |
A | Sandford-Holmes | Sheppard | Service | Fraser | 51.15 | FHSAA 4A District 8 |
| 2 | Riverview (Sarasota) |
A | Morris | Sanders | Sanders | Montas | 51.00 | FHSAA 4A District 8 |
| 3 | Dunbar |
A | Watson | Hopkins | Dudley | Gary | 48.26 | Louie Bing Invitational |
| 4 | Charlotte |
A | Aylesworth | Houston | Christensen | Hutto | 50.23 | |
| 5 | Palmetto |
A | Anderson | Conwell | Pompey | Hornes | 51.03 | 51.03 Manatee Hurricane Invitational |
| 6 | Venice |
A | Gonnelli | Evans | Luzzi | Williams | 51.50 |