| Rank | Name | Time | Year |
| 1 | Aubrey Runyon | 44.32 | 2025 |
| 2 | Brooke Coffman | 46.01 | 2023 |
| 3 | Morgan Goode | 46.65 | 2002 |
| 4 | Ally Parramore | 47.06 | 2016 |
| 5 | Chris Teike | 48.20 | 1986 |
| 6 | Denese Murphy | 49.20 | 1985 |
| 7 | Courtney Clarkston | 49.30 | 1997 |
| 8 | Michelle Cummings | 49.40 | 1995 |
| 9 | Casey Macharia | 49.63 | 2025 |
| 10 | Emma Beavins | 49.74 | 2018 |
| 11 | Morgan Sandrock | 49.96 | 2023 |
| 12 | Marla Isaac | 50.12 | 2026 |
| 13 | Lauren Sandrock | 50.28 | 2019 |
| 14 | Andrea White | 50.36 | 1997 |
| 15 | Hali Parker | 50.60 | 2009 |
| 16 | Stephanie Mariani | 50.91 | 1998 |
| 17 | LeeAnn Baker | 51.09 | 1987 |
| 18 | Sarah Cearing | 51.42 | 2005 |
| 19 | Brittany Ward | 51.44 | 2012 |
| 20 | Tina Spicer | 51.90 | 1994 |
| 21 | Jerica Richter | 52.31 | 2006 |
| 22 | Mary Kay Spicer | 52.80 | 1991 |
| 23 | Katie Gramse | 52.88 | 2002 |
| 24 | Samantha Woodall | 52.99 | 2019 |
| 25 | Carly Sandrock | 53.09 | 2015 |