| Lane | Team | Squad | Seed | Note | 
|---|---|---|---|---|
| 2 | Crooms | A | NT | |
| 3 | University (OC) | A | 3:43.23 | 3:43.23 Coach Ernest McPherson Invitational @ Sanford Seminole High | 
| 4 | Seminole (Sanford) | A | 3:30.00 | |
| 5 | Hagerty | A | 3:38.22 | |
| 6 | Lake Howell | A | DNS | 3:48.26 Coach Ernest McPherson Invitational @ Sanford Seminole High | 
| 7 | Geneva School | A | NT |