| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | AGARD, Donovan | University (Orlando) | NT | |
| 2 | NAPIER, Xavian | East River | NT | |
| 3 | MCKAY, Joshua | Geneva School | 1:01.91 | Highlander Classic 2019 |
| 4 | SAVILLE, Mathew | Lyman | 1:00.35 | Lake Mary Invitational 2021 |
| 5 | WHITE, Charles | Geneva School | 1:00.65 | Mount Dora Christian Invitational Meet #1 |
| 6 | MOYER, Mitchell | Geneva School | NT | |
| 7 | UGARTE, Amante | Ocoee | NT |
| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | VANEGAS, Cairo | Oviedo | 58.14 | Silverhawk Quint 2021 |
| 2 | SCHOENFELD, Lukas | Hagerty | 57.11 | |
| 3 | GRIGGS, JaSean | Oak Ridge | 56.40 | Cypress Creek (Orlando) Home Meet 2 |
| 4 | FREDERICK, Jeremy | Hagerty | 56.00 | |
| 5 | FREDELL, Jonah | Oviedo | 56.12 | Silverhawk Quint 2021 |
| 6 | SANTIAGO, Colin | East River | 56.63 | Metro East Championship 2021 |
| 7 | QUINTERO, KEVIN | Oak Ridge | 57.81 | Cypress Creek (Orlando) Home Meet 2 |
| 8 | ROSS, Jefferson | Lake Howell | 58.79 | SAC Freshman Sophomore Championships 2021 |
| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | PALERMO RUIZ, Anthony | Lake Howell | 55.13 | Trinity Prep Invitational 2020 |
| 2 | LALOY, Wenter | University (Orlando) | 54.78 | |
| 3 | STEPHENS, Justin | Seminole (Sanford) | 54.39 | Lake Brantley Quad 2021 |
| 4 | BONDZIE, Michael | Lyman | 54.11 | |
| 5 | WALKER, Zachary | Oak Ridge | 54.20 | |
| 6 | FARWIG, Ryan | Hagerty | 54.50 | |
| 7 | INGRAM, Tyreese | Seminole (Sanford) | 55.00 | |
| 8 | ESQUIVEL, Darren | Oviedo | 55.75 | Brian Jaeger Elite Classic 2021 |
| Lane | Athlete | Team | Seed | Note |
|---|---|---|---|---|
| 1 | BLAKE, Jalen | Winter Springs | 51.95 | Silverhawk Quint 2021 |
| 2 | MCDONALD, Marshae | Lake Howell | 50.70 | Silverhawk Open 2021 |
| 3 | COOPER LLL , John | Ocoee | 49.89 | |
| 4 | RACE, Jonathan | Lyman | 49.45 | Bob Mosher Wildcat Open 2020 |
| 5 | WHITE , Christopher | Ocoee | 49.86 | |
| 6 | LEWIS, Tyrique | Seminole (Sanford) | 50.01 | Lake Brantley Quad 2021 |
| 7 | MORRIS, DeRel | East River | 51.79 | FHSAA 3A District 6 |
| 8 | TAYLOR, Brandon | University (Orlando) | 53.11 |