Matlock twice lost concentration shortly before half-time to slip to their first Northern Premier League defeat of the season.
Connor Hughes on 43 minutes and Tony Gray in first-half stoppage time scored the goals for Warrington Town.
Yet they came as Matlock looked the better side without stretching home keeper Tony McMillan.
Matlock tried hard in the second half but, apart from a Dwayne Wiley header that struck the post ten minutes from time, they rarely looked like recovering a point.
Rhys Sharpe replaced Tyler Blake in the one change from the starting line-up that drew at Sutton Coldfield, meaning Ted Cribley again played a more central role after operating on the wing at Sutton.
Sharpe created Matlock’s first dangerous moment with a penetrating run down the left.
His fine cross was well cleared as Joe Doyle-Charles attempted to drive the ball goalwards.
Gladiators’ keeper Barnes held a bobbling low shot from Dylan Vassallo before the next chance for the visitors.
Josh Hine was booked for an elbow on Sharpe and from the free kick Nico Degirolamo’s header was deflected wide.
The chance signalled a good spell of pressure from Matlock. A Darnelle Bailey-King centre failed to drop for Cribley before another fine break from the pacy winger saw him cross invitingly again, but no-one was there to finish.
Although Barnes had to smother the ball at Gray’s feet, there was no indication that Warrington would take a grip on the game before Matlock contributed to their own downfall.
Hughes was not picked up as he broke through to slip a shot past Barnes for the opening goal.
With Matlock still reeling from that setback, they gave the ball away in midfield and a pass was threaded through for Gray to calmly fire past the keeper.
The visitors’ Shaun Harrad needed more support and service to prosper up front, although he was denied on 52 minutes when James McCarten headed his shot behind for an unproductive corner.
Warrington were efficient and uncomplicated as they looked to be in relative comfort in the second half.
The long throws of Hine created problems for the Gladiators defence and Barnes did well to block a Gray effort from one, but clear-cut opportunities were at a premium.
Harrad was off target before Wiley’s header beat McMillan but rebounded off his left-hand post with ten minutes to go.
Matlock needed that chance to go in to set up a possible grandstand finish. But Warrington closed them down quickly and played sensibly, Phil Baker doing well to intercept a pass bound for Tyler Blake to ensure a clean sheet for his side.
As at Sutton the previous Tuesday, Matlock gave away sloppy goals that cost them.
WARRINGTON TOWN: 1 Tony McMillan, 2 Liam Goulding, 3 Rory McKeown, 4 Brad Barnes (16 Caleb Richards, 76mins), 5 James McCarten, 6 Phil Baker, 7 Josh Hine, 8 Sean Williams, 9 Tony Gray, 10 Dylan Vassallo (14 Joe Mwasile, 72mins), 11 Connor Hughes (15 Abou Sanogo, 66mins). Subs not used: 12 Adam Carden, 17 Gerard Kinsella.
MATLOCK TOWN: 1 Phil Barnes, 2 Nico Degirolamo, 3 Jake Green, 4 Joe Doyle-Charles (12 Marc Newsham, 76mins), 5 Adam Yates, 6 Dwayne Wiley, 7 Darnelle Bailey-King (15 Luis Rose, 69mins), 8 Ryan Wilson, 9 Shaun Harrad, 10 Ted Cribley, 11 Rhys Sharpe (16 Tyler Blake, 66mins). Subs not used: 14 Callum Lloyd, 17 Zeyn Hakeem.
REFEREE: Nik Storey (Manchester).
GLADIATOR: Ryan Wilson