| Lane | Team | Squad | Athlete 1 | Athlete 2 | Athlete 3 | Athlete 4 | Seed |
|---|---|---|---|---|---|---|---|
| 2 | Huntington Local | A | Ater | Griffin | Hutton | Kifer | 58.80 |
| 3 | Paint Valley | A | Smith | McFadden | Vaughn | Dye | 56.00 |
| 4 | Unioto | A | Renner | Heath | Pelletiere | Debord | 52.90 |
| 5 | Southeastern | A | Barnes | Jones | Mitten | Jones | 55.58 |
| 6 | Adena | A | Hurless | McGarvey | Ogier | Thompson | 58.32 |
| 7 | Zane Trace | A | Henning | Alley | Corcoran | Perry | 1:05.00 |