| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 1 | Messiah |
A | 49.95 | Mason-Dixon Track & Field Invitational |
| 2 | California (Pa.) |
A | 49.00 | |
| 3 | Millersville |
A | 47.61 | PSAC Outdoor Track & Field Championships |
| 4 | Shippensburg |
A | 46.63 | 2023 Colonial Relays |
| 5 | Kutztown |
A | 47.50 | PSAC Outdoor Track & Field Championships |
| 6 | Cortland St. |
A | 48.66 | CNY College Challenge |
| 7 | Felician |
A | 49.18 | 2023 TCNJ Invitational |
| 8 | Clarion |
A | 50.57 | Tomcat Invitational |
| Lane | Team | Squad | Seed | Note |
|---|---|---|---|---|
| 2 | Maryland Club Running | A | 56.00 | |
| 3 | Gettysburg |
A | 51.93 | Mason-Dixon Track & Field Invitational |
| 4 | Marymount (Va.) |
A | 51.11 | George Mason Ebanks Invitational |
| 5 | PSU-Harrisburg |
A | 51.72 | 2023 Messiah Multi/Invitational |
| 6 | Juniata |
A | 54.20 | 2023 Messiah Multi/Invitational |