The Barcelona forward has failed to prove his fitness to 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."
Villa is Spain's all-time record goalscorer, having netted 51 goals in 82 appearances.
The push for more color on their cleats, adidas has added the infrared colorway to the adiZero f50 Line.
Ajax has released their away soccer jersey for the 2013/14 season. Get in the Gear.
adidas has added some color to the Predator line with this new colorway. Released on May 17. Be the first to own a pair.
The Liverpool 2013/14 soccer jersey is available for pre-order. The Reds are eager to compete for silver next season.
The adidas Nitrocharge is the most talked about new cleat in years. It was released on May 15 and will take your pitch by storm.
Chelsea won the Europa League on Wednesday. The Blues defeated Benfica 2-1 on injury time goal from Ivanovic.
The Juventus jersey for the 2013/14 was released earlier this week. Be the first to wear the colors of the Old Lady.
Bayern Munich are kings of Germany and hope to be crowned kings of Europe at Wembley later this month!
Borussia Dortmund hears Wembley Calling and are headed to London for the Champions League final.
The new PUMA evoSPEED 1.2 FG. A great new soccer cleat with classic look and first time for PUMA to be on the cleat.