| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed | Note |
|---|---|---|---|---|---|---|---|---|
| 1 | Johnstown | A | Thompson | Szabo | Thomas | Slack | 1:02.59 | |
| 2 | Utica | A | Dunn | Pryor | Morey | Heckman | 1:00.27 | |
| 3 | East Knox | A | Howard | Lilly | Lilly | Shelley | 58.63 | |
| 4 | Mar. Highland | A | Hinkle | Stephens | Williams | Schirtzinger | 55.80 | |
| 5 | Fredericktown | A | Smith | Tenney | Cockrell | Rine | 58.36 | |
| 6 | Columbus School for Girls | A | Moore-Utley | Jones | Behbehani | Seals | 58.63 | |
| 7 | Mount Gilead ms | A | Rizor | Nettle | Floyd | Bowie | 1:01.95 |