Real Sociedad pair Mikel Oyarzabal and Martin Zubimendi, along with Villarreal winger Alex Baena, have left the Spain squad following Friday night's 2-1 win in Denmark.
The trio will not travel to Tenerife for Monday's match against Switzerland, which has now lost significance as Spain has secured its place in the knockout stage of the UEFA Nations League, while Switzerland has no chance of finishing second.
Zubimendi and Baena both experienced physical issues during the second half of Friday's game and had to be substituted, with Zubimendi suffering a groin injury and Baena injuring his ankle.
The Spanish Football Federation (RFEF) explained, "With the next game so close, it has been decided to release both players to avoid any risks."
Oyarzabal, who scored Spain's opening goal in the game, also received a yellow card and is suspended for Monday's match. The RFEF has allowed him to return to his club, which is set to play a rescheduled Copa del Rey tie against fifth-tier side Jove Espanyol on Thursday night.
Spain coach Luis de la Fuente has called Atletico Madrid under-21 midfielder Pablo Barrios into the squad to provide cover after he played in the U-21s' 0-0 draw with England on Friday.