| Lane | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|
| 1 | YOUSSOUF, Iyman | Unattached | 57.00 | ||
| 2 | SMITH, Thomas | SO | St. Thomas (Fla.) |
55.89 | 55.89 (400m IH 55.89) STU UNDER THE LIGHTS OPEN |
| 3 | CHO-SAM, Zackary | FR | Southeastern U. |
54.50 | |
| 4 | WRIGHT , Rasmus | Unattached | 51.05 | ||
| 5 | THOMAS, Mario | SR | Southeastern U. |
54.20 | |
| 6 | JOHNSON JR., Cedric | SO | Southeastern U. |
54.50 | |
| 7 | CRABBE, Mkhori | FR | St. Thomas (Fla.) |
56.48 | 2025 97th Clyde Littlefield Texas Relays |
| 8 | DANIEL, Rick | SR | St. Thomas (Fla.) |
58.83 | UCF Black & Gold Challenge |
| Lane | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|
| 2 | FLOWERS, Ruben | SO | Barry |
NT | |
| 3 | MARTINEZ, Mario | SO | Florida National | 1:07.21 | Fr. Luis Ripoll S.J. Relays |
| 4 | JOHNSON JR, Tramaine | FR | Florida National | 1:01.72 | GMAC Championship |
| 5 | MERTILUS, Fodney | JR | Fort Lauderdale | 1:03.00 | |
| 6 | BROWN, Timothy | SO | Warner |
NT | |
| 7 | STEPHENSON, Demitri | FR | St. Thomas (Fla.) |
NT |