The officials held responsible for "undermining Ukraine's territorial integrity, sovereignty and independence" will be banned from travelling to the 28-nation bloc and will see their assets there frozen, the EU said in a statement.
The names of the individuals targeted were not immediately released but would be included in the official publication of the move in the bloc's legal journal early tomorrow.
The decision taken by the EU governments' ambassadors in Brussels brings the total number of Russians or pro-Russian individuals in Ukraine targeted by the EU's sanctions to 48.
The initial sanctions were adopted following Russia's annexation of Ukraine's Crimean Peninsula last month.
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The EU and the U.S. Now also accuse Russia of destabilising eastern Ukraine. NATO says Moscow has amassed some 40,000 troops just across the Ukrainian border and could invade the country within days if it wanted to.
The EU move comes after the US decided to broaden its own sanctions to include seven Russian government officials and 17 companies with links to President Vladimir Putin.
EU leaders have threatened Russia with tougher economic sanctions, for example targeting its financial industry or the energy sector, if the situation in eastern Ukraine further escalates, but those sanctions are not yet being considered.
No "tripwire" has so far been set to trigger those measures, British Foreign Secretary William Hague told lawmakers in London.