MPs have rejected a Labour amendment to the Government motion on Syria by a majority of 112 votes.
The amendment had called for military action to only be taken once the UN Security Council had voted in light of a report from weapons' inspectors on the ground in Syria.
The Prime Minister had earlier said the "abhorrent" chemical weapons attack in Damascus last week had caused "sickening human suffering" and could not be ignored.
But he stressed his plans should not be compared to the allied invasion of Iraq in 2003, which led to the downfall of Saddam Hussein.
Starting an emergency debate in the Commons, Mr Cameron admitted the earlier conflict had sparked "deep public cynicism" about military intervention.
However, he said: "This is not like Iraq, what we are seeing in Syria is fundamentally different. We are not invading a country. We are not searching for chemical or biological weapons."
MPs have begun voting on a Labour amendment to the Government motion on Syria, a move which will test Mr Cameron's authority.
He warned "decades of painstaking work" would be undone if there was no international action.
"The global consensus against the use of chemical weapons will be fatally unravelled, a 100-year taboo would be breached," he warned.
The Prime Minister admitted there was no "one smoking piece of intelligence" that made it 100% certain the Assad regime was behind the atrocity.
But he said he had been convinced by the available evidence and told MPs it was now up to them to make the same judgement.
He warned Syrian leader Bashar Assad would conclude he could "use these weapons again and again and on a larger scale and with impunity" if the world stood by.
"In the end we can't know the mind of this brutal dictator, all we can do is make a judgment about whether it is better to act or not to act and make a judgment about whether he is responsible or not responsible," he said.
Despite efforts to secure a UN Security Council resolution, Mr Cameron argued this should not be the only basis for possible action.
Indicating Britain could ignore a veto by Russia or China, he said his test would be if there was "overwhelming opposition" at the UN.
The debate came after Downing Street published its legal advice for action and a letter detailing the position of intelligence experts.
Government lawyers believe Britain could launch a targeted strike on humanitarian grounds without agreement at the UN.
And evidence from the Joint Intelligence Committee (JIC) found a chemical weapons did take place and it was "highly likely" the Assad regime was to blame.
MPs will vote tonight on the principle of military action, with any direct British involvement requiring a further Commons vote.
But the Prime Minister's hopes of a united political response have already been dashed after Labour vowed to oppose the Government motion.
Mr Cameron had already been forced to accept the need to give UN inspectors more time to report and MPs a further vote but this was still not enough to win round the Opposition.
It called the Government's new position "opaque" and tabled an alternative motion demanding "compelling evidence" the Assad regime was behind the atrocity.
Number 10 reacted furiously to Ed Miliband's positioning, accusing him of "playing politics" and attempting to divide the country.
But in the Commons, the Labour leader insisted "evidence should precede decision" and that the UN was not an "inconvenient sideshow" but essential to building a case.
He told the House he had not ruled out military intervention but said: "We need to be clear-eyed about the impact this would have."
Calling for respect to be shown to the UN, he added: "I am very clear about the fact that we have to learn the lessons of Iraq."
A frustrated Number 10 argued that the evidence from the intelligence services and publicly available material, including YouTube videos, was clear.
"Are we seriously suggesting that rows of three-year-old children frothing at the mouth and dead were somehow made to act this out?" a senior source said.
"Experts who have looked at this video have said it is conclusive that something like sarin was used here. The idea that this was concocted in some way is ridiculous."
Former foreign secretary Sir Malcolm Rifkind said it was impossible to have 100% proof the regime was involved but insisted the available evidence was strong.
The chemical weapons attack was in the same Damascus suburb where a sustained military attack by government forces was taking place, he pointed out.
And he warned MPs: "At this very moment, the Assad regime in Damascus is watching very carefully to see whether they will get away with what they have done.
"If there is no significant international response of any kind, then we can be absolutely certain that the forces within Damascus will be successful in saying we must continue to use these whenever there is a military rationale for doing so.
"There is no guarantee that a military strike against military targets will work, but there is every certainty that if we don't make that effort to punish and deter, then these actions will indeed continue."
UN weapons inspectors are due to finish their work on Friday and will report directly to secretary general Ban Ki-moon within 24 hours.
But their conclusions will not apportion blame - they will only set out the evidence on whether a chemical attack happened or not.
Former Tory Foreign Office minister David Davis said Britain could be "conned" into military intervention in Syria by rebels who were using chemical weapons to draw the West into the conflict.
He told the Commons MPs must have a justification for military action that is "not constructed" and said a "more aggressive" disclosure of intelligence could be necessary.
The former party leader candidate added there was no "clear moral imperative" to intervene in Syria.
He said: "There are many reasons for us to worry about this concern. We do not want to be conned into a war in effect by actions designed just to do that.
"There are plenty of facts around, or at least reported facts, reported that the UN representative for human rights in Syria thought there was concrete evidence of rebels having sarin gas."
Shadow transport spokesman Jim Fitzpatrick resigned his position after saying he would vote against Mr Miliband's policy.
He quit after telling the Commons he was "opposed to military intervention in Syria, full stop".
A senior Labour source said: "Jim Fitzpatrick has tonight handed in his resignation as shadow transport spokesman."
Before the debate in Britain, Syria's Parliament called on British MPs to avoid "reckless action", warning intervention could help terrorists and lead to the deaths of UK troops.
In a letter, it declared a strike would be "an aggressive and unprovoked act of war" and said: "We ask you not to bomb us but to work with us."
Assad also issued a fresh warning on Thursday that the country would "defend itself in the face of any aggression".
Six British RAF Typhoon jets have been sent to Cyprus as tensions mount, in what the Ministry of Defence called a "prudent and precautionary measure".
Meanwhile, reports suggested Russia - a key ally of Syria - is sending warships to the Mediterranean.
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