Sir Menzies said there was no "public appetite" for a vote
The Lib Dems have ended their campaign for an "in or out" referendum on UK membership of the European Union.
The pro-European party had argued that instead of a vote on the Lisbon Treaty a referendum was needed on Britain's broader relationship with the EU.
But ex-leader Sir Menzies Campbell said there was no "public appetite" for a vote now that the treaty was ratified.
He said it was now time to concentrate on making Europe work better post-ratification.
He told BBC Two's Daily Politics: "When the issue of whether or not there should be a referendum on Lisbon came up then we said it would be much more sensible to have a referendum on 'in or out' but the Lisbon Treaty is now history."
He added: "The one thing about the Lisbon Treaty is that there is no appetite of any kind whatsoever, for the foreseeable future, for any other treaty change in Europe.
"Now all of the effort and energy has got to be directed towards making Lisbon work and that's the real issue now."
But Lord Pearson, the new leader of the UK Independence Party, which campaigns for Britain's exit from the EU, said there "jolly well is an appetite" for a referendum among the public.
He told The Daily Politics: "We should have a referendum on in or out. I actually put an amendment down an in the Lords during the Lisbon proceedings in the hope that the Lib Dems would support it and, blow me down, they didn't."
On Tuesday, UKIP will launch a national campaign for a referendum on EU membership.
When he first called for an "in or out" referendum in 2007, Sir Menzies said the public deserved an "honest debate" on Europe.
His successor as Lib Dem leader, Nick Clegg, continued to campaign for such a vote, orchestrating a walkout of his MPs in March last year when the then Speaker Michael Martin refused to grant them a debate on Britain's membership of the EU.
But Conservative critics claimed the Lib Dems were not serious about the pledge and had only come up with it to "paper over" the party's own internal divisions on Europe.
A Lib Dem spokeswoman denied this, saying they had always seen a referendum on EU membership as a preferable option to a Lisbon Treaty vote.
She said: "We always said, if there was going to be a referendum, then there should be a referendum it should be on the larger question of Britain's relationship with the EU."
But she said it was a "completely different political landscape" to two years ago when Sir Menzies first made his pledge and as a "pro-European party" they wanted to concentrate on making the Lisbon Treaty work.
The Conservatives have ditched their commitment to a referendum on the Lisbon Treaty, following ratification but pledged to repatriate some powers from Europe and introduce a Sovereignty Act, to safeguard the supremacy of the British Parliament.
Shadow Europe minister Mark Francois denied claims by Sir Menzies that the Sovereignty Act would have no "practical constitutional effect" and was merely an example of "showboating" for party political purposes.
Mr Francois said he wanted Britain to emulate other EU states, such as Germany, which he said had laws guaranteeing sovereignty.