Juventus forward Alessandro Del Piero has been omitted from the Italy squad for Wednesday's friendly in Spain.
Del Piero has not featured for the Azzurri since he had a quiet game in September's Euro 2008 qualifier against France, which finished goalless.
But he has enjoyed a fantastic season in Serie A and had been hoping for a recall.
Alberto Gilardino, who like Del Piero was a member of Italy's World Cup-winning squad, is also facing a huge fight to be in the party for Euro 2008 after being left out by Roberto Donadoni.
There are no major surprises in Donadoni's 21-man squad, with striker Marco Borriello keeping his place after making his debut in February's 3-1 friendly win over Portugal.
Borriello, who has scored 17 league goals for Genoa this season, is expected to understudy Bayern Munich's Luca Toni in Austria and Switzerland in June.
Marco Materazzi is in line to make his first appearance for Italy since injuring his thigh in the 3-1 friendly defeat in Hungary last August.
Marco Amelia (Livorno), Gianluigi Buffon (Juventus); Andrea Barzagli (Palermo), Fabio Cannavaro (Real Madrid), Fabio Grosso (Olympique Lyon), Marco Materazzi (Inter Milan), Massimo Oddo (AC Milan), Christian Panucci (AS Roma), Gianluca Zambrotta (Barcelona); Massimo Ambrosini (AC Milan), Alberto Aquilani (AS Roma), Mauro Camoranesi (Juventus), Daniele De Rossi (AS Roma), Gennaro Gattuso (AC Milan), Simone Perrotta (AS Roma), Andrea Pirlo (AC Milan); Marco Borriello (Genoa), Antonio Di Natale (Udinese), Vincenzo Iaquinta (Juventus), Fabio Quagliarella (Udinese), Luca Toni (Bayern Munich).