The likelihood of the petition for a referendum being given the go-ahead was always unlikely, given the level of opposition in the Spanish Parliament.
The ruling conservative People's Party and main opposition group, the Socialists, as well as the centrist Union for Progress and Democracy (UPD) were all against the move.
The move was rejected by 299 members of parliament, or 86%, with just 47 voting in favour.
However, Artur Mas, the Catalan President, said he had expected the vote would be lost but insisted: "They can't block the will of the Catalan people."
The spectre of a breakaway Catalonia, which accounts for one- fifth of the Spanish economy and 16% of its population, has become a major headache for Prime Minister Mariano Rajoy, who is battling high unemployment and the scars of a deep recession.
Earlier this year, in a motion brought by the UPD, MPs blocked the Catalan government's independence drive, voting by a large majority to reject Catalonia's "right to decide".
Mr Mas has made clear that his next step will be to defy the Parliament in Madrid and go ahead with the referendum anyway.
He has already set the date of November 9, just two months after the independence referendum vote in Scotland, which is being closely watched in Catalonia.
Mr Rajoy has warned he will use the courts to block the Catalan government from holding a vote, even though Mr Mas argues that, if it is a non-binding consultation, then it should be legal.
However, he has also signalled he will not break the law. So if the referendum is shut down by the courts, the Catalan President is expected to use the next election in Catalonia, which must be held by 2016, as a proxy vote on independence.
During the debate yesterday, he said: "I defend that Catalonia should remain in Spain because I can't conceive of Spain without Catalonia nor of Catalonia outside of Spain and Europe," he told the assembly. Together we all win and separate we all lose."
Opinion polls show around half of Catalans support independence, yet a much higher number want the right to vote on the matter.
Catalonia - the land of artists Joan Miro and Salvador Dali and architect Antoni Gaudi - is home to some of Spain's biggest companies, including banks Caixabank and Sabadell, global infrastructure company Abertis and utility Gas Natural.
Catalan business leaders have been cautious about taking sides on independence, fearing a backlash. However, the head of Spain's largest pharmaceutical company, Barcelona-based Grifols, broke the silence last week when he backed Mr Mas's drive to hold a referendum.
Meanwhile, the independence cause in Canada suffered a major blow when the anti-separatist Liberal Party won a majority government in provincial elections, thus eliminating any chance of a new referendum in Quebec for several years.
Pauline Marois, the Parti Quebecois leader, announced she would resign as party chief. Liberal leader Philippe Couillard will now replace her as premier of the province.
Alistair Carmichael, the Scottish Secretary, said the Canadian result provided Scotland with an interesting lesson: that politicians must accept results and move on. A "neverendum", he insisted, "should never be an option".