| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 1 | Booker | A | Brewer | Bradley | Behn | Forrester | NT |
| 2 | Manatee | A | Jean | Bean | Stewart | Sanders | 3:59.00 |
| 3 | Palmetto | A | Davis | Martin | Carter | Hughes | 3:40.00 |
| 4 | Bayshore | A | Anders | Lyons | Boyer | Barnes | 3:30.29 |
| 5 | Southeast | A | Thomas | Davis | Thomas | Schmucker | 3:35.02 |
| 6 | Braden River | A | Brown | Wolfe | Stephens | Campbell | 3:44.21 |
| 7 | Parrish | A | Rosario | Giles | Eisch | Kelley | NT |