The defending champions were aiming to become the first nation to retain the title since Brazil in 1962. But, after losing their opening match 5-1 to the Netherlands, they surrendered their crown in a cauldron of noise in the Maracana.
First-half goals from Eduardo Vargas and Charles Aranguiz did the damage as Spain failed to contain the rampant Chileans, or muster a response of any real substance.
Their elimination means they are the fifth defending champions to fail to progress from the group stage at the World Cup.
It was Chile's first competitive victory over Spain - and just reward for an attacking masterclass.
Questions will inevitably be asked about the international futures of Spain coach Vicente Del Bosque and several of his most trusted players, such as goalkeeper Iker Casillas and midfielders Xavi and Xabi Alonso.
There was little sign of shock as the full-time whistle sounded; more despondency and resignation. Chile celebrated, but not wildly; they deserved their victory and will not want to stop there.
Spain must now contest a dead rubber against Australia on 23 June, while Chile and Netherlands meet in an encounter that will determine who finishes top of Group B.
Chile were tipped by many to do well in the competition, but few predicted they would administer the fatal blow to Spain and one of the game's most glittering eras.
Their fans arrived in huge numbers and helped generate an incredible pre-match atmosphere, although a group of them storming the media centre before kick-off provided a sour preamble.
They seemed to enjoy far greater representation inside the ground, and the noise levels rocketed as Jorge Sampaoli's team made an encouraging start - Alonso forced into a vital goalmouth clearance before Gonzalo Jara headed narrowly wide.
Spain were narrow victors when the sides met at the 2010 World Cup, and again in a friendly the following year, but needed a last-minute equaliser to snatch a draw last September.
Chile have been dubbed by some the 'Spain of South America' - a reference to the Spaniards in peak form - and their pace, power and pressing is a sight to behold.
They opened the scoring in style after Alonso carelessly under-hit a pass, allowing Alexis Sanchez to begin a scintillating move that ended with Aranguiz squaring for Vargas to round Casillas and slot home.
Spain responded positively, and predictably enjoyed plenty of possession, but Alonso shot way over the bar and Diego Costa fired wide.
Del Bosque made a massive call by relegating Xavi to the substitutes' bench, moving David Silva into his attacking midfield position and using the younger, quicker Pedro on the right.
But, if anything, Spain looked worse without a man who, while enduring a poor campaign and struggling in Friday's thrashing by Netherlands, has for so long been La Roja's outfield leader, the symbol of their philosophy and one of the best players in the world.
The dropping of centre-back Gerard Pique was less of a surprise but, similarly, did not benefit Spain and their defence looked no more assured or unified with Javi Martinez partnering Sergio Ramos.
They needed to get to half-time without conceding again, but even that proved beyond the European champions as Casillas palmed a Sanchez free-kick straight out to Aranguiz, whose control and finish with the outside of his foot was of the highest order.
Spain finally rallied after the break, Andres Iniesta sending Costa clean through - but Mauricio Isla arrived from right-back with a perfectly-timed challenge. Sergio Busquets then missed an open goal following Costa's overhead kick.
Isla should have extended Chile's lead only to sky a shot from close range, and at the other end Iniesta and substitute Santi Cazorla were twice denied by Claudio Bravo.
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