| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Clearwater |
A | Benjamin | Chambers | Walker | Gray | NT | 3:39.78 Pinellas County Athletic Conference |
| 2 | St. Petersburg |
A | Andringa | Lopez | Wu | Delbrune | 3:36.20 | 3:36.20 Mustang Invitational |
| 3 | Lakewood |
A | Morris | Wooten | Slocumb | Dent | 3:27.29 | FHSAA 2A Region 3 |
| 4 | Boca Ciega |
A | Eubanks | Malone | Price | Marthaller | 3:33.32 | FHSAA 2A District 11 |
| 5 | Gibbs |
A | Ferguson | Roberson | Delgado | Danford | 3:38.25 | 3:38.25 Thomas Jet Jackson Invitational |
| 6 | Largo |
A | NT |