| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Winter Park |
A | Stevens | Stryku | Westfall | Clark | 8:25.80 | 8:25.80 Metro East Championship |
| 2 | Bishop Moore |
A | Solby | Fasen | Voss | Bisigni | 8:32.67 | 8:32.67 2024 Spring Break Bash |
| 3 | Tavares |
A | Mars | Ludwig | Mewborne | Hartley | 8:36.51 | FHSAA 3A District 5 |
| 4 | Edgewater |
A | Casanas | Cleveland | Collin | Vecchio | DNS | FHSAA 3A District 6 |
| 5 | McIntosh | A | Nowell | Will | Gray | Voigt | 9:00.00 | |
| 6 | University (Orlando) |
A | Ramirez | Maurer | Clarke | Diaz-Garcia | 9:05.73 | 9:05.73 Lake Mary Invitational 2024 %28Regional Qualifer%29 |
| 7 | Windermere Prep |
A | Hollingshead | Mansilla | Hardoon | Guzman | 9:15.12 | |
| 8 | Orlando Christian Prep |
A | windbush | Kaufman | Verkennes | uscanga | 9:40.89 | 9:40.89 Mount Dora Christian Invitational %232 |
| 9 | Jones |
A | Hall | Hall | Horne | Thomas, jr | NT |