Needing a point to reach consecutive European Championships, a combination of steely focus, last-ditch blocks and some Spanish profligacy meant the home side were scoreless before Alvaro Morata’s ghosting run and header deep into the second half.
Minutes earlier Scott McTominay had triggered revelry among the Scots as he lashed a wonderful free-kick past Unai Simon, only for the goal to be ruled out after a VAR review.
Scotland’s anguish was compounded in the closing moments as Spain grabbed a second via a Ryan Porteous own goal.
Scotland, who lost captain Andy Robertson to injury in the first half, sit top of Group A, three points above their hosts, and will qualify for Euro 2024 on Sunday if Norway fail to beat the in-form Spanish.
Or, if needed, they can get the job done next month when they travel to Georgia for their penultimate game.
Scotland left the pitch to a rapturous reception from a huge travelling support in Spain. This had all the hallmarks of a famous night, only for it to be laced with the harshest of sucker punches.
Scotland’s challenge at the start of the evening? Take a point, or hope Norway would drop the ball in Cyprus. The latter always looked unlikely – and so it proved as Norway eased to a 4-0 win – and from kick-off in the sweltering Seville heat, the heft of the task facing the Scots was evident.
Spain are far from the husk of a side who wilted at Hampden in March. Barcelona’s Ferran Torres steered a shot wide when through on Angus Gunn after just a minute.
A series of corners somehow flew across the Scotland goal, which was starting to lead a charmed life. None more so than when Mikel Merino’s slashed shot hit a post, flew between goalkeeper and line, before spinning out.
Alvaro Morata’s opener stung Scotland after Scott McTominay’s free-kick was ruled out
The booming gasp which met that Merino chance was loud, but nothing compared to the roar midway through the second half courtesy of McTominay.
On a rare chance for a Scotland shot at goal, the Manchester United midfielder, the thorny thistle in the La Roja side in March, whipped a glorious free-kick high into the far corner from a tight angle.
But jubilation turned to jaw-smacking agony as VAR intervened and the goal was ruled out, initially, it seemed, for a foul by Jack Hendry on the goalkeeper.
That was what the flashed up on the screens in the stadium, but Uefa later said it was because Hendry was offside and interfering with play.
Confusion reigned and the debate about that whole process will linger unless and until Scotland have booked their place in Germany.
It changed the game, and on 73 minutes Morata’s drifting run pierced the Scotland defence, leaving Gunn helpless.
Substitute Che Adams prodded at Simon when he should have done better late on, and that profligacy was punished as Porteous slid the ball into his own net after an Aaron Hickey slip caused panic at the back.
Scotland are far from done in this run, but this one will sting for some time.
