Colliery had enjoyed an impressive 3-0 win over Cleethorpes in the previous round.
But a higher ranked Wakefield side proved to be too strong and ran out comfortable winners at the Jones Recreation Ground on Sunday.
Danielle Thompson netted Colliery’s consolation goal in front of a vocal crowd of 144.
Harworth play in the Sheffield & Hallamshire Women & Girls League – alongside Wakefield’s development team.
Trinity currently lead the Premier Division of the North East Regional Womens Football League.