| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 2 | Danville | A | Hackman | Mickley | Meade | COLOPY | 1:40.72 | Millersport HS Laker Invitationa |
| 3 | Centerburg | A | Joiner | Marshall | Rhodebeck | BALL | 1:40.38 | Bob Royer Invitational |
| 4 | Fredericktown | A | Anderson | Rine | Roush | Rojas | 1:32.77 | Fredericktown Invitational |
| 5 | East Knox | A | Springer | Proudfoot | Madden | Clark | 1:40.00 | Jim Proper Invitational |
| 6 | Centerburg | B | Clayton | Plagge | Schaller | Runyon | DQ | |
| 7 | Fredericktown | B | Alexander | Hogg | Smith | McManis | NT |