Atletico Madrid winger Vitolo, who is on loan at Las Palmas until January, has been a key player for Spain since Julien Lopetegui took charge just over a year ago, scoring four times in five games in qualifying.
“Yesterday morning in training, the player suffered a strain in the medial collateral ligament of his knee,” the Spanish Football Federation (RFEF) said in a statement.
“After consultation with the medical services of his club and those of the RFEF, the player will leave the squad this evening.”
Vazquez has three international caps, but his only appearance in a competitive fixture came when Italy beat Spain 2-0 at Euro 2016.
Spain will be looking for revenge when they host the Azzurri at Real’s Santiago Bernabeu on Saturday with both sides locked together on 16 points at the top of Group G of European qualifying.
La Roja travel to Liechtenstein on September 5.
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