Manchester United midfielder Juan Mata has been left out of the Spain squad for the World Cup qualifiers against Italy and Albania in October.
Mata, 28, was omitted from the Euro 2016 squad before being recalled by new Spain manager Julen Lopetegui.
However, Mata was an unused substitute in the 2-0 friendly win in Belgium and the 8-0 World Cup qualifying win over Liechtenstein this month.
Chelsea midfielder Cesc Fabregas, 29, is again omitted from the squad.
He was not included in Lopetegui's first squad.
West Ham keeper Adrian, who has conceded 16 goals in six Premier League games this season, is also left out.
Chelsea striker Diego Costa, Manchester United goalkeeper David de Gea and Manchester City pair David Silva and Nolito are the four Premier League players in the 23-man squad.
Spain, who beat Liechtenstein 8-1 in their opening Group G game, play Italy in Turin on 6 October and Albania in Shkoder on 9 October.
Goalkeepers: David de Gea (Manchester United), Pepe Reina (Napoli), Sergio Rico (Sevilla).
Defenders: Nacho Fernandez (Real Madrid), Jordi Alba (Barcelona), Dani Carvajal (Real Madrid), Sergio Ramos (Real Madrid), Gerard Pique (Barcelona), Javi Martinez (Bayern Munich).
Midfielders: Koke (Atletico Madrid), Sergi Roberto (Barcelona), Saul Niguez (Atletico Madrid), Sergio Busquets (Barcelona), David Silva (Manchester City), Lucas Vazquez (Real Madrid), Thiago Alcantara (Bayern Munich), Andres Iniesta (Barcelona), Isco (Real Madrid).
Forwards: Jose Callejon (Napoli), Vitolo (Sevilla), Alvaro Morata (Real Madrid), Diego Costa (Chelsea), Nolito (Manchester City).