Villa has failed to prove his fitness to coach Vicente del Bosque after fracturing his tibia while in action for the Catalan side at the World Club Challenge in Japan in December.
Del Bosque had promised to give the 30-year-old until the last moment to get himself ready but, with his squad set to be named on Sunday, he has called time on Villa's race to be fit.
A statement on the Spanish Football Federation's official website tonight read: "David Villa cannot definitely be on the list for Euro 2012.
"Villa spoke with the national team doctor this afternoon after the arrival of the team in the Austrian town of Schruns.
"After not getting in the final practice and development desired, David Villa spoke with Del Bosque, who relayed that although it was Villa's wish to be with the team in Poland and Ukraine, that his physical state was 'not as good' as would be desirable."