| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Trinity Christian (Deltona) |
B | Morales | Robin | Pardilla | Sanchez | NT | |
| 4 | Geneva School |
B | Patenaude | Bradley | Guedes | Trifone | NT | |
| 5 | Lake Mary |
B | Mcclelland | Keller | Gerboc | Kalia | NT |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Father Lopez |
B | Aldrich | Green | Kealy | Breive | 1:02.00 | |
| 3 | First Academy (Orlando) |
B | Pine | Lozada Da Silva | Robinson | Herrington | 51.03 | |
| 4 | Trinity Prep |
A | DeBaise | Hickok | Osleger | Bennett | 55.00 | |
| 5 | Trinity Christian (Deltona) |
A | Rodriguez | Easter | Miller | Marrow | 57.38 | Mount Dora Christian Invitational Meet #2 |
| 6 | Hagerty |
B | Smith | Jones | Leger | Ruping | DQ | |
| 7 | Bishop Moore |
B | Dorcely | Ongaro | Myers | Elena Padin Castillo | NT |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Cornerstone Charter |
A | Gainex | Hierbe | Johnson | Janis | 53.64 | Freedom Last Chance 2025 |
| 2 | Geneva School |
A | Flores | Roberts | Lindsay | Duvra | DNF | 50.55 Mount Dora Christian Invitational Meet %232 |
| 3 | Lake Howell |
A | Ramon | Bibb | Bibb | Stokes | FS | FHSAA 4A Region 1 |
| 4 | Lake Mary |
A | Foster | Holley | Smith | Carter | 49.58 | 49.58 Hagerty Track and Field Invitational 2024 |
| 5 | Hagerty |
A | Trapaga Morejon | Farwig | Burke | Jaime | 49.76 | |
| 6 | Bishop Moore |
A | Suffran | Sullivan | Maddalena | Sweeney | DNF | FHSAA 2A Region 2 |
| 7 | Father Lopez |
A | Bridger | Bernal | Leiva | Lancing | 51.68 | FHSAA 1A Region 2 |
| 8 | First Academy (Orlando) |
A | Awosika | Claytor | Schmid | Spafford | 51.16 | Citrus League HS Championship 2025 |