| Lane | # | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|---|
| 1 | 664 | VEGA RAMOS, Darwin | SO | Lake-Sumter State | 56.99 | UCF Knights Invite |
| 2 | 130 | CASTRO, Paolo | FR | Saint Leo |
56.75 | Tampa Spartans Open |
| 3 | 89 | BROWN, Timothy | JR | Warner |
55.13 | Spartan Relays |
| 4 | 81 | GOSSETT, Daishaun | JR | Warner |
52.61 | Sun Conference Championship 2025 |
| 5 | 173 | BESSI, Flippo | SO | Florida Tech |
53.00 | |
| 6 | 129 | WARREN, Joshua | FR | Saint Leo |
55.34 | Tampa Spartans Open |
| 7 | 174 | ESTEBANES, Diego | SO | Florida Tech |
56.83 | Sunshine State Conference Track Championship |
| 8 | 295 | SCRABIS, Grant | JR | Embry-Riddle |
57.39 | Embry-Riddle Eagle Invite |
| Lane | # | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|---|
| 1 | 792 | MARTIN-O'KELLY, Erick | FR | Barry |
NT | |
| 2 | 567 | HINTON, Thomas | JR | Husson |
1:08.51 | 2025 NAC Track & Field Championship |
| 3 | 651 | CASSAMAJOR, Marven | FR | Florida National | 58.40 | GMAC Championship |
| 4 | 793 | JIMENEZ, Kruz | JR | Barry |
57.99 | Nova Southeastern University Alumni Invitational |
| 5 | 345 | DESIMONE, Luke | FR | Embry-Riddle |
58.00 | |
| 6 | 775 | WHITE, Khaleb | SO | Barry |
1:01.08 | Sunshine State Conference Track Championship |
| 7 | 631 | WILSON, Jahlandon | SO | University of The Bahamas | NT |