| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 3 | Wharton |
A | Polk | Grady | Robinson Bey | Bataille | 54.36 |
| 4 | Strawberry Crest |
A | Keuchler | Georges | Perez | Starford | 54.32 |
| 5 | Jefferson |
A | Bradford | Ellis-Williams | Pitcher | Nunn | 54.33 |
| 6 | Freedom (Tampa) |
A | Kolar | Daniels | Wood | Hernandez | 55.39 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 1 | Middleton |
A | Pena | Swint | Moore | Smiley | 54.15 |
| 2 | Robinson |
A | Jones | Velazquez Martinez | Arons | Lleras | 52.35 |
| 3 | Alonso |
A | Lauricella | Sanchez | Turtle | Lachapelle | 51.20 |
| 4 | Bloomingdale |
A | Exil | Hayes | Courtney | Courtney | 50.98 |
| 5 | Durant |
A | McGritt | Crouch | Howard | Knighten | 51.19 |
| 6 | Blake |
A | Shaw | Page | Lue | Blyden | 51.71 |
| 7 | Chamberlain |
A | Nash | Lucas | Mosley | Nash | 53.81 |
| 8 | Gaither |
A | Chandler | Kindred | Ozuna | Jones | 54.24 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 1 | Leto |
A | Allen Chona | Garcia | Singh | Singh | 49.81 |
| 2 | Hillsborough |
A | GRISSETT | Lovett | Eugene | Carriera | 48.46 |
| 3 | Sumner |
A | Brown | Bright | Rankin | Barton | 47.98 |
| 4 | Tampa Bay Tech |
A | Cineas | Thomas | George | Long | 47.45 |
| 5 | Lennard |
A | Jorden | Johnson | McDaniel | Bradshaw | 47.61 |
| 6 | Newsome |
A | Alaalaoui | Dawson | Browne | Ramos | 48.00 |
| 7 | Armwood |
A | Ashley | Harris | Johnson | Harper | 49.49 |
| 8 | Spoto |
A | Debose | Joseph | Hatter | Snead | 50.70 |