The two remaining BetVictor Northern Premier clubs both lost in their second round games on Saturday.
FC United were thumped 7-0 by National League leaders Barrow, who had seen off NPL side Atherton Collieries in their first round replay on Tuesday night.
There was no shame in defeat for the Reds, as Barrow also plundered seven in their last league game.
Matlock Town put up more of a fight, as they were beaten 2-1 at National League side Eastleigh.
Two lapses of concentration a minute either side of half-time proved their undoing, despite Marcus Marshall’s second half goal threatening to earn them a replay.
Cav Miley struck the Spitfires’ opening goal with virtually the last kick of the first half, and within a minute of the restart, Lewis Beale made it 2-0.
Marshall gave the Gladiators 21 minutes to find an equaliser, but it wasn’t to be - even though Jon Stewart saved a 74th minute penalty to keep the tie alive.
Matlock were backed by more than a hundred travelling supporters in a crowd of 718.
The draw for the next round of the FA Trophy takes place on Monday.
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