More than 140 people were turned away from the Queensland border in the first five hours after it closed to anyone from New South Wales and the ACT.
Those travelling from Victoria are also among the 14 million Australians now barred from entering the state as it attempts to contain the spread of coronavirus.
It comes as the state government confirmed there were no new virus cases on Saturday, and there remains a total of 11 active cases in the state.
Police said delays of more than 90 minutes to cross checkpoints were reported in the lead-up to the 1am Saturday morning closure as Queenslanders rushed to return home.
There were 142 people turned away at the border between 1am and 6am because they did not meet the requirements to enter.
Chief Superintendent Mark Wheeler, speaking to reporters near the Coolangatta checkpoint, said that number included 18 Queenslanders, who will now incur a “considerable cost” getting home.
Queensland residents returning from the declared hotspot locations must now arrive by air and pay $2800 for two weeks of hotel quarantine.
Residents of communities along the border will be eligible for a pass to cross in and out of Queensland, but must not travel beyond the local area.
However the chief superintendent acknowledged there would still be people in “those little catch-22 situations”.
“What they would need to do is look at their circumstances, look at what passes are available, and then, if necessary, make an application to the Chief Health Officer for an exemption,” he said.
The state’s deputy premier, Steven Miles, said the state had done 157,000 coronavirus tests in the past fortnight, with seven of those coming back positive.
“Queenslanders continue to come forward in droves to be tested for this virus,” he said.
In contrast, Victoria recorded 466 new cases on Saturday, while New South Wales reported nine.
Chief Superintendent Wheeler said efforts to stop the spread mean entering Queensland is now “the exception rather than the rule” and there will be consequences for anyone caught flouting the system.
“Those people who lied on their border declaration passes, and we‘ve charged a number of them, they’ll find themselves before court,” he said.
“Instead of getting an on-the-spot fine of over $4000, they‘ll find themselves in court, facing a fine of up to $13,000 and/or six months imprisonment.
“It‘s a really serious offence.”