Secretary of State Mike Pompeo has said the United States will never recognise Russia's annexation of Crimea, and will continue to insist that Ukraine's territorial integrity be restored.
In a statement released by the State Department, Pompeo said the US will hold to its long-standing principle of refusing to recognise Kremlin claims of sovereignty over territory seized by force, in violation of international law.
He called for Russia to respect principles and "end its occupation of Crimea".
The statement was released shortly before Pompeo was due to testify before the Senate Foreign Relations Committee, where he is expected to face tough questioning about President Donald Trump's summit with Russian leader Vladimir Putin.
Trump has previously suggested that US opposition to Russia's annexation of Crimea could be reconsidered.
In anticipation of potentially hostile questions, Pompeo's Crimea Declaration sought to underscore US commitments to the post-Second World War international order that many believe Trump has weakened.
In it, Pompeo took Russia to task for its actions in Ukraine, particularly the 2014 annexation of Crimea.
Russia has defended its move, saying the annexation was approved by Crimean voters in a referendum.
Foreign ministry spokeswoman Maria Zakharova dismissed the comments on her Facebook page.
"We know the worth of such momentous declarations," she wrote.
The US and its European allies have all said the referendum was illegal as it was held without the consent of the government in Kiev and was conducted in a highly flawed manner.
"Through its actions, Russia has acted in a manner unworthy of a great nation and has chosen to isolate itself from the international community," Pompeo said.
Even before the statement was released, he was taking a tough line on Russia and defending the Trump-Putin meeting as he prepared for his Senate testimony.
He also reiterated US support for an investigation that held Russia responsible for downing a Malaysian airliner over eastern Ukraine in 2014. Pompeo said what the Russians did was "deeply immoral" and that those responsible for the MH17 disaster should be held accountable.
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