The Spanish Football Federation has confirmed that David Villa will not be fit to take up a place in the holders' squad for Euro 2012.
Barcelona striker Villa has not played since December after suffering a broken leg against Al Sadd in the Club World Cup.
Spain coach Vicente del Bosque was prepared to give the 30-year-old as much as time as possible to prove his fitness for this summer's European Championship.
However, after the squad arrived in Austria for a pre-tournament training camp, Villa spoke with Del Bosque to confirm he would not be ready to play in Poland and Ukraine.
A statement on the Spanish Football Federation's official website 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."
Villa wrote on Twitter: "I've tried, but I can't be 100% to play the Eurocup. I called Vicente del Bosque to say him. It's the honest thing I could do.
"Thanks to all of you for your support. I will continue working to return to play as soon as possible with Barca and Spanish National Team."
Del Bosque is due to name his final 23-man squad for Euro 2012 on Sunday and must decide which strikers to rely on in the next few weeks.
Fernando Torres was added to the squad on Monday, along with Chelsea team-mate Juan Mata, after showing signs of returning to form in recent weeks.
The loss of Villa, who was part of the Euro 2008 and 2010 World Cup-winning squads, is another blow for Spain as club team-mate Carles Puyol is also out due to injury.