| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | First Academy (Orlando) |
B | Herrington | Adams | Visser | Visser | NT | |
| 4 | Geneva School |
B | Patenaude | Guedes | Bradley | White | NT | |
| 5 | Trinity Christian (Deltona) |
B | Kriner | Robin | Pardilla | Sanchez | NT |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Victory Christian (LL) |
B | Choquette | Hunt | McInnes | Brangers | 1:53.00 | |
| 3 | Lake Mary |
B | Sackllah | Martinez | Williams | Chester | NT | |
| 4 | Cornerstone Charter |
A | Loiseau | Tibo | Baselyous | Hurlock | NT | |
| 5 | Bishop Moore |
B | Dorcely | Ongaro | Myers | Elena Padin Castillo | DNS | |
| 6 | Geneva School |
A | Lindsay | Flores | Newhartz | Duvra | NT | |
| 7 | Hagerty |
B | Geter | Smith | Moise | Leger | 1:52.00 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Trinity Christian (Deltona) |
A | Rodriguez | Easter | Miller | Marrow | NT | |
| 2 | First Academy (Orlando) |
A | Claytor | Schmid | Spafford | Awosika | 1:51.00 | |
| 3 | Victory Christian (LL) |
A | Kratky | Barroso | Lemmond | Williams | 1:45.00 | |
| 4 | Lake Mary |
A | Sigurdsson | Foster | Smith | Holley | 1:20.00 | |
| 5 | Hagerty |
A | Trapaga Morejon | Farwig | Burke | Jaime | 1:44.34 | |
| 6 | Father Lopez |
A | Bridger | Bernal | Lancing | Aldrich | 1:50.00 | |
| 7 | Lake Howell |
A | Neal | Verdu | Springmann | Scott-Bernard | 2:04.05 | Charlie Harris Relay |
| 8 | Bishop Moore |
A | Cantero | Maddalena | Sullivan | Sweeney | NT |