| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 3 | St. Francis Borgia | A | Charboneau, Macey | Comage, Bernadette | Garcilaso, Shekinah | Kimminau, Danni | 55.14 |
| 4 | Francis Howell Central | A | Cooper, Rebecca | Clark, Daisy | Ebert, Lauren | Hanna, Shante | 53.54 |
| 5 | Troy-Buchanan | A | Morice, Amanda | Edwards, Adene | Matech-Edwards, Salena | Brooks, Kaitlyn | 53.94 |
| 6 | St. Francis Borgia | B | Brinker, Samantha | Rembecki, Ana | Williams, Allison | Maune, Liz | 1:02.14 |