Enrique and Alba suffered a reported breakdown in relationship during his spell at the Blaugrana and it appears to have continued since being announced as La Roja boss.
Alba has now been snubbed for a second consecutive squad, while Atletico Madrid's Koke returns after his sublime 2018/19 season form.
Isco, Dani Carvajal. Diego Costa and Sergi Roberto all miss out through injury.
Wolves defender Jonny Otto, on loan from Atletico, received his first international call-up.
While there were returns to the squad for Real Betis defender Marc Bartra and Barca forward Paco Alcacer - currently enjoying a loan spell with Borussia Dortmund.
Spain squad in full:
Goalkeepers: David De Gea (Manchester United), Kepa Arrizabalaga (Chelsea), Pau Lopez (Real Betis)
Defenders: Cesar Azpilicueta (Chelsea), Raul Albiol (Napoli), Nacho, Sergio Ramos (Real Madrid), Marcos Alonso (Chelsea), Gaya (Valencia), Marc Bartra (Real Betis), Jonny Otto (Wolverhampton)
Midfielders: Sergio Busquets (Barcelona), Rodrigo, Saul, Koke (Atletico Madrid), Thiago Alcantara (Bayern Munich), Dani Ceballos (Real Madrid)
Forwards: Marco Asensio (Real Madrid), Alvaro Morata (Chelsea), Suso (AC Milan), Rodrigo Moreno (Valencia), Iago Aspas (Celta Vigo), Paco Alcacer (Borussia Dortmund)
Spain play Wales in a friendly in Cardiff on October 11 before facing England in the UEFA Nations League on October 15 in Seville