| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Ridge Community | A | Formey | Glasgow | Jean Baptiste | Goins | 54.10 | |
| 2 | George Jenkins | A | Antoine | Wilson Cole | Alexander | Wright | 52.00 | |
| 3 | IMG Academy | A | Walker | Saucedo Franco | Albee | Baker | 51.12 | |
| 4 | Jones | A | White | Burton | Blake | Townsend | 46.39 | 2019 FHSAA State Track & Field Championships |
| 5 | Lakeland | A | Wright | Jimenez | Wills | Dewsey | 48.79 | 2019 FHSAA State Track & Field Championships |
| 6 | Haines City | A | Boyke-Johnson | Kerr | Cruzado | Roberts | 51.86 | 51.86 Polk County Championship 2019 |
| 7 | Kathleen | A | Wilson | Williams | Fritz | Patton | DNS | |
| 8 | Lake Region | A | Eddins | Groomes | Pita | Dunlap | 54.10 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Frostproof | A | Campbell | Brown | Whalen | Cobb | DQ | |
| 4 | Goleman | A | wright | AGUIAR | Rodriguez | ROSE | NT | |
| 5 | Riverview | A | Neptune | Cesar | Gordon-Watkins | Ceelen | NT | |
| 6 | Tenoroc | A | Ellington | Mcclurg | Wesley | Bell | NT |