| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Albright |
A | Maley | Alverado | Hammond | Zamichieli | 3:40.43 | Purple & Gold Open |
| 2 | A | Perrelli | Passarelli | Evely | Sorrentino | 3:31.50 | 2025 Kings College Monarch Open | |
| 3 | York (Pa.) |
A | Apsley | Moore | Bonham | Diallo | 3:28.47 | York College Twilight |
| 4 | A | Nonez | Anderson | Valera | Castelli | 3:24.62 | Danny Curran Invite | |
| 5 | Hood |
A | Barberena | Seskar | Mins | Ellington | 3:25.58 | 2025 Messiah Multi/Invitational |
| 6 | DeSales |
A | McCulloch | Guthier | Marsh | Callano | 3:30.12 | Muhlenberg Invitational |
| 7 | Stevens |
A | Robinson | Bindra | Jutras | Savino | 3:36.50 | Muhlenberg Invitational |
| 8 | Alvernia |
A | Anderson | Wendel | Downs | St. Pierre | 3:42.11 | York College Twilight |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Stevenson |
A | Howard JR | Reimert | Hammond | McCann | 3:23.17 | York College Twilight |
| 2 | Widener |
A | McNear | Celestin | Rudd | Knowles | 3:22.21 | 2025 Paul Donahue Invitational at Widener University |
| 3 | Eastern |
A | Davis | Walton | Greenwood | Ugorji | 3:20.62 | Hopkins Loyola Invitational |
| 4 | Misericordia |
A | Billings | McGuffey | Kieffer | Hunter | 3:19.65 | Washington & Lee Carnival |
| 5 | Delaware Valley |
A | O'Dunlami | Troy | Madison | Middel | 3:20.43 | Penn Relays |
| 6 | Messiah |
A | Baxter | Sarkett | Condon | Xenakis | 3:21.79 | Penn Relays |
| 7 | Lebanon Valley |
A | Goundie | Bruchak | Di Cesare | McKell | 3:22.94 | York College Twilight |
| 8 | A | Gorski | Petty | Klem | Novrocki | 3:24.43 | 2025 Kings College Monarch Open |