Jordi Alba and Koke have been left out of Luis Enrique's first Spain squad, while Alvaro Morata and Sergi Roberto have been recalled.
Barcelona full-back Alba has been overlooked by his former club coach in favour of Valencia's Jose Gaya and Chelsea's Marcos Alonso, both of whom missed out on a place at the World Cup.
Alba fell out of favour with Luis Enrique during his final season at Barca, and the defender admitted he felt the coach had little confidence in his ability.
He was a favourite of Julen Lopetegui, though, and played all four of Spain's matches at the World Cup even after Lopetegui was sacked for holding talks with Real Madrid without the Royal Spanish Football Federation's knowledge.
Koke played in both of Atletico Madrid's first two LaLiga games of the season, but the midfielder was criticised in some quarters for his performances at the World Cup.
Only 13 of the 23 players who went to Russia have retained their places. Of the other 10, Andres Iniesta, David Silva and Gerard Pique have retired from international football, while Pepe Reina, Alvaro Odriozola, Nacho Monreal, Alba, Koke, Lucas Vazquez and Iago Aspas have been left out.
Morata has earned a recall, though, having started the new season under Maurizio Sarri at Chelsea in encouraging fashion, with one goal scored in three appearances.
Barca utility player Roberto is back in the squad, as is AC Milan forward Suso and Athletic Bilbao's Inigo Martinez. There are also call-ups for Atleti midfielder Rodrigo, Real Madrid's Dani Ceballos, Real Betis' uncapped goalkeeper Pau Lopez and Napoli defender Raul Albiol, who turns 33 on Tuesday and last played for his country in March 2015.
Spain face England and Croatia in their opening two UEFA Nations League matches on September 8 and 11 respectively.
Spain squad in full:
David de Gea (Manchester United), Kepa Arrizabalaga (Chelsea), Pau Lopez (Real Betis); Dani Carvajal (Real Madrid), Cesar Azpilicueta (Chelsea), Raul Albiol (Napoli), Diego Llorente (Real Sociedad), Nacho Fernandez (Real Madrid), Sergio Ramos (Real Madrid), Inigo Martinez (Athletic Bilbao), Marcos Alonso (Chelsea), Jose Gaya (Valencia); Sergio Busquets (Barcelona), Sergi Roberto (Barcelona), Rodrigo (Atletico Madrid), Saul Niguez (Atletico Madrid), Thiago Alcantara (Bayern Munich), Dani Ceballos (Real Madrid); Isco (Real Madrid), Marco Asensio (Real Madrid), Alvaro Morata (Chelsea), Diego Costa (Atletico Madrid), Suso (AC Milan), Rodrigo Moreno (Valencia).
