Spain coach Luis de la Fuente on Sunday called up centre-back Robin Le Normand to his Nations League squad following an injury to Eric Garcia ahead of the world champions' 3-2 win away to England.
Garcia had initially been picked in the starting line-up at Wembley but pulled out at the last moment, to be replaced by Marc Pubill.
In a statement, the Spanish football federation said that Garcia "felt some discomfort in his left knee and, after undergoing tests, it was decided to withdraw him from the squad for the upcoming internationals, to avoid any risks".
De la Fuente had said after Saturday's victory at Wembley that Garcia's problem was "nothing serious".
The world and European champions host Croatia in Seville on Tuesday in their next match.
French-born Le Normand has not played for Spain in almost a year.
He was part of the squad that won the Euros in 2024 but was not selected for the World Cup earlier this year.