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First half goals from Kevin Gameiro and Rodrigo Moreno ended Ernesto Valverde's chances of a second successive domestic double since taking over at the Camp Nou in 2017.
Barcelona started the game slowly, and Valencia almost took the lead after just five minutes, but Gerard Pique made a brilliant goal line clearance from Rodrigo, following an error from Clement Lenglet.
However Marcelino's side found a breakthrough on 20 minutes, as French international Gameiro slotted home from close range after a flowing Valencia counter attack.
Los Che doubled their lead on 34 minutes, as Spanish forward Rodrigo powered home a header from Carlos Soler's whipped cross from the right wing.
Skipper Lionel Messi hit a post just after the hour mark, before the Argentina international netted his 51st goal of the season on 73 minutes, turning home from a yard out after Lenglet's header hit the post.
