US Attorney General Eric Holder and British Home Secretary Theresa May told an international conference on asset recovery that those responsible would be held accountable.
"There should be no mistake, we are determined in our efforts to be successful," Holder told a press conference in London at the start of the two-day forum.
"We are determined to hold accountable those who were responsible for the theft of these Ukrainian assets and we are also determined to ensure that those assets are returned to the Ukrainian people."
He expected the eventual total to amount to tens of billions of dollars.
Makhnitskyi described the Yanukovych regime as an "organised criminal group" whose tentacles reached throughout the administration.
"The new government was set up and we found that our treasury was empty and the funds were misappropriated," he said.
Yanukovych was ousted in February following a series of massive protests after he decided to scrap an agreement with the European Union in favour of closer ties with Russia.
Ukrainian Interior Minister Arsen Avakov, senior government officials, prosecutors and representatives from financial centres and international organisations around the world are also attending the conference, Britain's Home Office said.
Britain's Serious Fraud Office announced on the eve of the conference that it had launched a money-laundering investigation into possible corruption in Ukraine and frozen USD 23 million in assets.
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