Thiago Alcantara, Fabian Ruiz and Diego Llorente also missed out on the squad, while surprise inclusion Abel Ruiz started in the front three in their 4-0 victory over Georgia last time out and West Ham United midfielder Pablo Fornals made an appearance off the bench.
Left-back Jose Gaya was forced off through injury in that game, and has been withdrawn from the squad for Wednesday's match as a result, with Tottenham Hotspur full-back Sergio Reguilon arriving to battle with Jordi Alba for a spot in the back four.
Centre-back Aymeric Laporte was also brought off last time out, but that change was said to be precautionary regarding fatigue, rather than the suspected adductor issue, meaning the Manchester City man could be fit to stay in the squad.
Luis Enrique may be unlikely to risk his star defender for the final game though, with Raul Albiol or Inigo Martinez potentially filling in alongside Eric Garcia.
Brighton & Hove Albion goalkeeper Robert Sanchez was handed his Spain debut last time out, as he came on for Unai Simon in the final 15 minutes, and he could be set for his first start if Enrique rotates his stoppers.
At the other end of the pitch, Manchester City winger Ferran Torres should continue to occupy an attacking spot, having already impressed for the national team with nine goals in 19 appearances at the age of 20, while Alvaro Morata could return to lead the line after he was an unused substitute last time out.
Spain possible starting lineup: Sanchez; Azpilicueta, Garcia, Albiol, Alba; Llorente, Rodri, Soler; Torres, Morata, Sarabia