| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 1 | The Village School of Naples | B | Hills | Mauceri | Osthoff | Dasgupta | NT |
| 2 | Seacrest Country Day |
A | Tiafierro | Davies | Whitlow | Hitchon | 1:00.16 |
| 3 | First Baptist (Naples) |
A | Weissman | Lugo | Solano | Antonelli | 58.73 |
| 4 | Evangelical Christian |
A | Crocker | Deleon | Bennett | Edick | 50.41 |
| 5 | Babcock | A | DePue | Gonzalez | Boone | Marquez | 52.71 |
| 6 | The Village School of Naples | A | Michie | Josephs | Hartman | Osthoff | 1:00.00 |
| 7 | Evangelical Christian |
B | Marshall | Brown | Chapman | Sahadeo | NT |