The Hampshire County Women’s League top flight club entered the tournament for the first time in their 16-year history.
Drawn at home to higher division Poole Town, Widbrook were beaten 7-2 in the second qualifying round in a tie played on the artificial surface at Horndean Technical College.
Widbrook consolations came from Sharna White and Tia Bramble.
AFC Portchester and United Services Portsmouth were also knocked out at the same stage.
The Royals went down 3-2 at home to Badshot Lea while USP were beaten 2-0 at home by Eastleigh in the Community.
Pictures by Mike Cooter