Barcelona's Jordi Alba has been dropped from Spain's squad ahead of the Nations League fixtures against England and Croatia.
The 29-year-old left-back, who has 66 international caps, joins Atletico Madrid midfielder Koke and Celta Vigo striker Iago Aspas in being omitted from Luis Enrique's first 24-man squad.
Enrique took up the role of Spain manager in July after a poor World Cup campaign - led by Fernando Hierro after Julen Lopetegui was sacked on the eve of the tournament - which saw the team crash out to hosts Russia in the last 16.
What is the UEFA Nations League?
Andres Iniesta, David Silva and Gerard Pique - who helped Spain win the World Cup in 2010 - all retired from international duty in the summer following the tournament.
Enrique, who managed Barcelona between 2014 and 2017, has been trusted to turn around the fortunes of the national side after leading the Catalan club to two La Liga titles, three Copa del Reys and a Champions League trophy during his tenure.
"This is a special day for me," Enrique said. "To make the first list of players [as coach] has involved many hours and a lot of hard work.
"I understand there might be a lot of interest in players who are not on the list, that's normal, but I have nothing to say about those who are not on the list. I'm full of reasons to bring those 24 who are here."
Spain face England on September 8 at Wembley in the newly-formed Nations League - the Three Lions' first game since their World Cup semi-final run in Russia - before hosting Croatia three days later on September 11.
"These two games are against two semi-finalists from the last World Cup and it's going to be very important to see where we stand and how hungry we are," Enrique added, before quashing rumours over a reported bust-up with captain Sergio Ramos.
"I never said Sergio Ramos is 'just one more player'," he said.
"But this is my style and I didn't think it was necessary to speak to the player. Maybe he is, in your opinion, and I respect that, but there's no way he's just one more player.
"He's the captain, the second player in history who has played the most matches with this shirt [the Spain national shirt]. He could never be just one more player. I never said that.
"I can't wait to see them [the players]. I can't wait to see Sergio Ramos, to see them all, and to enjoy this really exciting national team for me."
Full Spain squad
Goalkepeers: David de Gea, Kepa Arrizabalaga, Pau Lopez
Defenders: Dani Carvajal, Nacho, Sergio Ramos, Diego Llorente, Inigo Martinez, Jose Gaya, Cesar Azpilicueta, Marcos Alonso, Raul Albiol
Midfielders: Sergio Busquets, Sergi Roberto, Rodrigo, Saul Niguez, Dani Ceballos, Thiago Alcantara
Forwards: Isco, Marco Asensio, Diego Costa, Rodrigo Moreno, Alvaro Morata, Suso