Arsenal returned to domestic competition with a bump as Blackburn overcame the Gunners to leapfrog them into fifth position in the table.
Bellamy celebrates Blackburn's winner
The visitors looked a different side from the heroic team that beat Real Madrid on Tuesday.
And Rovers took the lead on 18 minutes when Craig Bellamy fooled Philippe Senderos and crossed for Morten Gamst Pedersen to drive home from close in.
Arsenal raised the tempo late on but failed to break Blackburn down.
After their stunning victory at the Bernabeu, Arsenal were keen to carry their momentum through to league matters.
But Blackburn denied the visitors any time or space on the ball as the Gunners struggled to find their rhythm.
And it was the home side, revitalised after their sunshine break in Dubai, who took the lead on 18 minutes.
A cute trick from Bellamy took the Welshman past Senderos and his low cross was forced home by Pedersen from three yards.
The Norwegian's elation turned to despair soon after as he limped off with a hamstring strain, sustained as he fired Blackburn ahead, and he was replaced by Sergio Peter.
The home side seized control of the game and Andy Todd's backwards header exposed the frailties in the Arsenal back-line.
With the Gunners defence in disarray, Robbie Savage could have doubled Rovers' lead but he guided Steven Reid's low drive over the bar in a first-half dominated by the home side.
Arsenal introduced Robert Pires in place of Jose Antonio Reyes at the interval in a bid to inject some creativity into the game.
But Blackburn continued where they left off in the first half and hurried the Gunners into mistakes with Jens Lehmann's frustration earning him a yellow card for a push on Reid.
Bellamy always looked dangerous while at the other end, Arsenal's hero on Tuesday night Thierry Henry, was out of sorts and never threatened the Blackburn defence.
But Arsenal raised their tempo midway through the second half as they went in search of the equaliser.
Senderos headed just over from Henry's corner and Cesc Fabregas shot tamely at Brad Friedel.
Friedel was given a sterner examination when Emanuel Adebayor found himself unmarked from point-blank range, only for the American to thwart his close-range effort.
Gunners boss Arsene Wenger threw on Italian striker Arturo Lupoli for his Premiership debut, but Blackburn held out for a deserved three points to move into a Uefa Cup qualifying spot.
Blackburn boss Mark Hughes:
"It was a fantastic performance from each and every one of my players.
"We were really brave in everything that we did, we stuck to our task and caused them untold problems at the back, they really couldn't cope.
"They had a little bit of pressure towards the end of the game, but never really troubled Brad Friedel. We were excellent from start to finish today."
Arsenal boss Arsene Wenger:
"I'm very sad because I don't think we deserved to lose this game.
"For maybe five or six minutes in the first half we suffered a little bit, but overall we were always on top of the game but we couldn't score.
"We lacked a little bit mental sharpness maybe because of Tuesday night but overall we gave absolutely everything physically until the last second of the game."
Blackburn: Friedel, Neill, Nelsen, Todd, Gray, Reid, Savage, Tugay (Bentley 68), Pedersen (Peter 20), Bellamy, Sinama Pongolle (Mokoena 74).
Subs Not Used: Enckelman, Kuqi.
Booked: Todd, Peter, Bentley.
Goal: Pedersen 18.
Arsenal: Lehmann, Eboue, Toure, Senderos, Flamini, Fabregas, Diaby (Hleb 63), Silva (Lupoli 82), Reyes (Pires 45), Henry, Adebayor.
Subs Not Used: Almunia, Djourou.
Booked: Toure, Lehmann.
Ref: U Rennie (S Yorkshire).