Alvaro Morata's diving header and a Ryan Porteous own goal earned Spain an important 2-0 win over Scotland in Euro 2024 qualifying on Thursday.
Steve Clarke's side left furious after Scott McTominay scored a brilliant free-kick to put Scotland ahead but the goal was disallowed for a foul on Spain goalkeeper Unai Simon.
Group A leaders Scotland would have sealed qualification for the tournament in Germany next summer with a point in Seville.
Should second-place Spain avoid defeat against Norway, third, on Sunday, Clarke's team will reach consecutive European Championships.
Spain, three points behind Scotland, can wrap up their own qualification if they beat Norway, who thrashed Cyprus 4-0 on Thursday.
Scotland arrived in Andalucia with an immaculate record from their five previous qualifiers, including a 2-0 win over Spain in Glasgow in March.
It was Spain coach Luis de la Fuente's second game in charge and he insisted his team were in far better shape ahead of the rematch, and so it proved.
Thousands of visiting fans jeered Spain midfielder Rodri Hernandez, who had labelled Scotland's tactics and approach at Hampden Park "rubbish".
Rodri and Spain had a lot of touches, dominating possession and the game, with Ferran Torres firing wide in the first minute when he should have hit the target.
Mikel Merino struck the post and Alvaro Morata had a goal ruled out for offside and sent another effort wide as De la Fuente's side sought to break the deadlock.
Scotland left-back Andy Robertson was celebrating winning his 67th international cap, taking him into the country's top 10, but his night ended in disappointment when he was taken off with a shoulder problem following a collision with Simon.
Spain threw on winger Bryan Zaragoza for his debut at half-time and he instantly caught the eye, cutting inside from the left and firing narrowly over.
McTominay, who netted both of Scotland's goals against Spain in Glasgow, thought he had struck again when his stinging free-kick from an impossibly tight angle ripped into the far top corner, sparking wild celebrations.
However, the referee ruled out the goal after VAR instructed him to watch a replay, apparently for a foul by Jack Hendry on Simon, even though very little contact was made.
Scotland were still seething when La Roja broke the deadlock, Jesus Navas crossing for Morata to head home brilliantly, flinging himself at the ball to divert it past Angus Gunn after 73 minutes.
Spain's definitive second came in the final stages when the otherwise superb Aaron Hickey slipped, allowing Joselu to slide the ball across the box and Porteous bundled the ball into his own net while attempting to stop debutant Oihan Sancet from reaching it.
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