| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Leo | A | Early | Merida | Smith | Treesh | NT | |
| 4 | Ft. Wayne Canterbury | A | Kim | O'Connor | Erb | Cvetkoski | 1:44.17 | Trine HSR Qualifier #2 |
| 5 | Elkhart Christian | A | Curtis | Lanninga | Ware | Gentz | 1:42.21 | Trine HSR Qualifier #1 |
| 6 | Ft. Wayne Blackhawk Christian | A | Grimes | Fritcha | Weigand | Mitchell | 1:42.51 | Trine HSR Qualifier #1 |
| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 3 | Angola | A | Yankowiak | Grunner | Phillips | Tynan | 1:39.22 | 2025 Carmon Cripe Relays @ Northridge |
| 4 | Huntington North | A | McQueen | Kowalski | Stults | Mitchell | 1:37.80 | |
| 5 | Fairfield | A | Miller | Grotrian | Tsybulnik | Herschberger | 1:34.55 | 2025 Lancer Relays |
| 6 | Jimtown | A | Fleming | Inbody | Ambriz | Coughenour | 1:36.00 |