Full-back Jonny May led the way with a double, while England wing Charlie Sharples and Fijian flanker Akapusi Qera also crossed and fly-half Freddie Burns booted 14 points.
Four-time European champions Toulouse, though, still progress to the quarter-finals as Pool Six winners after Harlequins lost by a point against Connacht in Galway.
Wing Timoci Matanavou touched down twice for the visitors, while skipper Thierry Dusautoir scored early on and fly-half Lionel Beauxis added three conversions and a penalty.
But they were outplayed by a Gloucester team that despite already being eliminated from the quarter-final equation, blew them apart.
Young backs like May, Sharples and Burns had a field day, performing with a thrilling sense of adventure and creativity that had Toulouse's stamp all over it.
Gloucester delivered comfortably their best performance of the season, claiming a bonus point in the process, although it was too little too late in terms of this season's Heineken Cup following home and away defeats to Quins after losing in Toulouse.
For their part, Toulouse will limp, rather than swagger into the last eight for a 13th time, and Gloucester's performance exposed weaknesses in their armoury under pressure.
The Aviva Premiership club just needed to see off a Connacht side who had not won in any competition since September to book their spot in the quarter-finals.
The visitors dominated possession and territory throughout the match, but could not turn their pressure into points amid cold, wet and blustery conditions in Galway.
Niall O'Connor kicked all of Connacht's points while Quins, who still have a faint hope of qualifying as one the best runners-up, scored a try through Sam Smith and a penalty via the boot of Nick Evans.
Current Pool Four leaders Ulster are also guaranteed a spot in the quarter-finals with Toulouse after Friday night's action