| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 1 | Donahue Academy |
B | Minter | Schroeder | Freeze | Watson | 5:25.00 |
| 2 | Naples Christian Academy | A | Castro | Morisson | Alonso | Simon | NT |
| 3 | St. Elizabeth Seton School | A | Scuderi | Conlon | Cronin | Kleiber | NT |
| 4 | St. Anns | A | Day | Bijou | Wasser | Burke | 5:00.00 |
| 5 | Donahue Academy |
A | Graham | Singla | Lanham | Webb | DNS |
| 6 | Royal Palm Academy | A | Waller | O'Malley | Mulligan | Schuetz | 6:30.00 |
| 7 | Village School - Lower School | A | Michie | Rose | Mendenhall | Josephs | 6:30.31 |
| 8 | Naples Christian Academy | B | Weng | Meistrell | Snyder | Leeper | NT |