Berkhamsted were knocked out of the Premier Division Cup as they suffered a 2-1 defeat at the hands of Ampthill Town on Tuesday night.
Ampthill took the lead when Tyrone Taylor beat his man to a cross and headed home past Berko keeper Carl Tasker.
The Comrades fought back, though, and Bekim Culaj equalised when he spotted a gap in the defensive wall and struck a 20-yard free kick into the net on the half hour mark.
Ampthill regained the one-goal advantage in the second half, when Tasker saved a shot and full-back Paul Ericson pounced to ram home the loose ball.
Town keeper Jamie Buick then produced the save of the night to keep out a shot by Dan Griggs and see his side into the quarter-finals.
Berko boss Mick Vipond will be looking for a quick response from his side, with the Comrades in action against Southend Manor in the FA Carlsberg Vase on Saturday.
Berko will fancy their chances of beating the Essex Senior Football League Premier Division side, who have had a mixed start to the season.
They currently sit in 14th position in the league table and booked their place in the second round of the Vase with a 2-0 win at Haverhill Rovers.
Berko, meanwhile, saw off Hadley 2-0 in the first round to make it to this stage of the competition.
Saturday’s clash kicks-off at Broadwater at 3pm.