Qualifier Winners gets Preferred Lanes (4-5-3-6)
| Lane | Athlete | Yr. | Team | Seed | Note |
|---|---|---|---|---|---|
| 1 | HAMILTON, Rohan | SR | Northeast (OP) | 50.42 | q |
| 2 | ADEGBORIOYE, Adefemi | SR | Western | 49.20 | q |
| 3 | DEAS, Dewante | SR | Ely | 50.04 | Q |
| 4 | MACK, King | SO | St. Thomas Aquinas | 48.94 | Q |
| 5 | ROBERTS, Tajay | SR | Dillard | 49.62 | Q |
| 6 | CANELON, Eric | JR | Flanagan | 50.86 | Q |
| 7 | BOLDEN, Xavier | SR | Plantation | 49.94 | q |
| 8 | MORGAN, Shamar | SO | St. Thomas Aquinas | 50.80 | q |