| Time | Date | Name | Meet |
|---|---|---|---|
| 48.99 | Feb. 11, 2023 | Richie Kurlich | North Coast Athletic Conference Championships |
| 49.54 | Jan. 28, 2023 | Richie Kurlich | Denison vs. Ohio Wesleyan vs. Youngstown State |
| 49.78 | March 7, 2021 | Noah Houskeeper | Denison vs. Ohio Wesleyan |
| 50.52 | Feb. 15, 2020 | Noah Houskeeper | North Coast Athletic Conference Championships |
| 50.78 | Feb. 17, 2018 | Kenny Fox | North Coast Athletic Conference Championships |
| 51.56 | Feb. 11, 2017 | Bebe Wang | North Coast Athletic Conference Championships |
| 52.48 | Jan. 8, 2017 | Jack Lindell | Denison vs. DePauw |
| 52.50 | Jan. 30, 2016 | Jack Lindell | Denison vs. Ohio Wesleyan |
| 53.35 | Jan. 31, 2015 | Alex Nennig | Ohio Wesleyan vs. Denison |
| 53.80 | Oct. 12, 2013 | Jack Lindell | Denison vs. Toledo, Wabash |
| 54.42 | Jan. 13, 2012 | Spencer Fronk | Wittenberg vs. Denison |
