Manchester City duo Aymeric Laporte and Rodri are included in Spain's World Cup squad, but Liverpool midfielder Thiago Alcantara and Manchester United goalkeeper David de Gea miss out.
De Gea, who has 45 caps, has not made an international appearance for two years, while Thiago earned the last of his 46 caps at Euro 2020 last year.
There is also no place in the squad for Paris St-Germain defender Sergio Ramos.
Chelsea's Cesar Azpilicueta is among five Premier League players included.
Brighton's Robert Sanchez and Brentford's David Raya are also in the squad, but fellow goalkeeper Kepa Arrizabalaga of Chelsea misses out.
Barcelona forward Ansu Fati has been named by boss Luis Enrique despite being sidelined for much of the last two years through injury.
Athletic Bilbao winger Nico Williams, who only made his international debut in September, will also travel to Qatar along with former Manchester City forward Ferran Torres and ex-Chelsea striker Alvaro Morata.
Spain are in Group E alongside Germany, Costa Rica and Japan.
The 2010 World champions take on Costa Rica in their opening match on 23 November.
Spain squad
Goalkeepers: Unai Simon (Athletic Bilbao), Robert Sanchez (Brighton), David Raya (Brentford).
Defenders: Cesar Azpilicueta (Chelsea), Dani Carvajal (Real Madrid), Eric Garcia (Barcelona), Hugo Guillamon (Valencia), Pau Torres (Villarreal), Aymeric Laporte (Manchester City), Jordi Alba (Barcelona), Jose Gaya (Valencia).
Midfielders: Sergio Busquets (Barcelona), Rodri Hernandez (Manchester City), Gavi (Barcelona), Carlos Soler (Paris St-Germain), Marcos Llorente (Atletico Madrid), Pedri Gonzalez (Barcelona), Koke Resurreccion (Atletico Madrid).
Forwards: Ferran Torres (Barcelona), Nico Williams (Athletic Bilbao), Yeremi Pino (Villarreal), Alvaro Morata (Atletico Madrid), Marco Asensio (Real Madrid), Pablo Sarabia (Paris St Germain), Dani Olmo (RB Leipzig), Ansu Fati (Barcelona).