| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Olympic Heights |
A | Wattenmaker | Spindola | Rose | Herrera | 1:00.76 | Boynton Beach Home Meet |
| 3 | Seminole Ridge |
A | Juntunen | Worrell | Coffey | Davies | 52.66 | Welington Meet #1 |
| 4 | Park Vista |
A | Robinson | Fleuricin | Colt | Hernandez | 50.09 | Dr. Garcia Meet #3 2026 |
| 5 | Dr. Joaquin Garcia |
A | Pierre | Dominguez | Massena | Michel | 51.38 | Dr. Garcia Meet #2 |
| 6 | Somerset Canyons |
A | Occean | Jean Mary | Henry | Miller | 54.96 | St Andrews Varsity 3 |