In blustery conditions on the all weather surface at Belper Leisure Centre, the Nailers were missing new signing Mia Carnelley through injury but welcomed back top scorer Alice Faulding.
The opening stages of the match were even and Faulding linked up well with Sammy Bennett on a couple of occasions but Belper couldn't quite force the opening they were after.
Wirksworth seized the initiative and took the lead on 17 minutes. Belper were unable to clear a set-piece and Romy Shepherd was on hand with a close range finish. Shepherd had scored at the Raygar Stadium in the league fixture in early October and in truth was the best player on the pitch in this Cup encounter as well.
Within a minute they had doubled their lead when Poppy Lloyd produced a neat finish from a tight angle. Another scorer from the league encounter coming back to haunt the Nailers defence.
On 20 minutes came Belper's best chance so far when Lorraine Taylor spilled a cross but Bennett couldn't quite get her shot away.
Three minutes later Shepherd struck a sweet shot from outside the box but Emma Varnam was able to watch it go wide of her post.
The 24th minute saw Belper right back in the tie. A great through-ball from Bennett found India Amans and her shot looped up off Taylor in to the goal to make it 1-2.
Five minutes later Varnam produced a great save to keep the Nailers within touching distance.
It was a quiet start to the second half and the wind picked up noticeably, making it difficult to judge anything aerial.
Hannah Dutton replaced the hard working Jess Needham and impressed.
In the 56th minute, Belper got their first chance of the half with Bennett's shot from range and moments later she displayed great footwork to create a chance for herself, the shot was just wide.
In between these two chances for Bennett, Sarah Stimpson crashed a shot off the post at the other end but Belper were enjoying the better of the match and searching for an equaliser.
Nine minutes from time Faulding tried her luck from range but Taylor was equal to it.
The last couple of chances went to the visitors, Shepherd had a great chance to seal the tie on 85 minutes when played in but her shot was wide.
On 87 minutes the tie was killed off when Belper only half cleared a Wirksworth attack and Grace Bradbury fired home from outside the box to make it 1-3.
Dom Anderson's side gave a really good account of themselves once again - in the league, the Colts are 4 points off top with 3 games in hand and represented another really big test.
The Nailers drop in to the County Plate so there is still hope of silverware for the ever improving Ladies Team.
Belper Town: Varnam, Pickard (C), Powles, Roberts, Cowan, Bennett, Needham, Worrall, Faulding, Buxton, Amans.
Subs: Whysall, Layton, Dutton, Macluskey, Jackson.
Referee: Rhys Watkinson