| Lane | Team | Squad | Seed | Note | 
|---|---|---|---|---|
| 1 |  Apopka | 
		A | NT | |
| 2 |  Lake Highland Prep | 
		A | 53.05 | |
| 3 |  Olympia | 
		A | 51.41 | Windermere Meet of Champions Classic 2024 | 
| 4 |  Wekiva | 
		A | 49.87 | Olympia Titans Invitational | 
| 5 |  Horizon | 
		A | 51.05 | Windermere Meet of Champions Classic 2024 | 
| 6 |  Foundation Academy | 
		A | 52.67 | Windermere Meet of Champions Classic 2024 | 
| 7 |  Winter Park | 
		A | 53.50 | |
| 8 |  Windermere Prep | 
		A | DNF |