| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Glades Day |
A | Treto | Martinez | Hall | Soriano | NT | |
| 3 | John I. Leonard |
A | Augustin | lapointe | garcia-molina | betemit | 4:27.63 | John I Leonard Invitational |
| 4 | Pahokee |
A | Johnson | Bythwood | White | Woodson | 3:56.04 | FHSAA 1A Region 4 |
| 5 | Somerset College Prep | A | Bourne-Morgan | Wright | Rodgers | Miller | 4:25.20 | South Fork Meet #2 2024 |
| 6 | Glades Central |
A | Sellers | Frazier | Kyles | Cruz | DNS | Anquan Boldin Invitational 2024 |