Alexis Sanchez and Chile will not be in attendance at the 2018 World Cup after losing 3-0 to Brazil in their final group game.
Lionel Messi inspired Argentina came from behind to beat Ecuador 3-1, allowing them to leapfrog Chile up into the automatic qualification spots.
Chile were 3rd going into the final round of matches but Colombia, Peru and Argentina all climbed above them on Tuesday night.
Beaten to the 5th play-off spot on goal difference, Chile coach Juan Antonio Pizzi immediately announced his resignation following the failed qualification campaign.
"I take full responsibility," Pizzi told reporters. "We gave everything but it was not enough. I will not continue."
They haven't qualified for a World Cup since 1982 but Peru's 1-1 draw with Colombia secured 5th spot and they will face New Zealand in a two-legged play-off.
How Ospina's incompetence for Colombia cost Chile dear
Arsenal goalkeeper David Ospina has totted up a variety of howlers during Colombia's World Cup qualifying campaign.
His teammate Alexis Sanchez won't be able to look him in the eye for some time following his ridiculous mistake against Peru on Tuesday night.
Jose Pekerman's Colombia were leading 1-0 through James Rodriguez and that scoreline would've been enough to secure Chile 5th spot above the Peruvians.
However, Peru were awarded an indirect free-kick in the 76th minute and Paolo Guerrero bizarrely decided to shoot anyway.
Ospina should've tipped it round the post after diving to save it - although ideally he would've just let it go in and the referee would've awarded Colombia a free-kick.
The free kick would not have counted if Arsenal's 29-year-old back-up keeper didn't palm it into his own goal.
Ospina is probably glad the goal didn't cost Colombia their place at the World Cup but he definitely won't be looking forward to being reunited with Sanchez in London.
Veteran Arsenal boss Arsene Wenger appears to have made the former Nice number one his cup goalkeeper for the season ahead.
The out-of-sorts Colombian stopper failed to keep a clean sheet in the Europa League wins over Cologne and BATE Borisov.