“He signed it yesterday,” said State Department spokesperson Jen Psaki.
Obama wasn’t expected to sign the controversial sanctions into law until later this week.
The legislation, Ukraine Freedom Support Act of 2014, sailed through Congress despite criticism it could send mixed messages to U.S. allies.
Critics have argued the legislation doesn’t take into consideration ongoing negotiations between Moscow and European powers over its alleged involvement in Ukraine’s internal conflict.
The Russian government says its only involvement in the conflict is humanitarian in nature, though Kiev claims Moscow is propping up separatist forces in Ukraine’s east.
Russia warned earlier this month new U.S. sanctions could be met with counter measures from Moscow.
Previous rounds of sanctions have contributed to a collapse in the value of the Russian rouble. The value of the Russian currency has halved in the past 12 months, and was continuing to fall on Wednesday.
However, the conflict in Ukraine is also dragging down the world economy, according to Russian presidential spokesperson Dmitry Peskov.
“When we don’t see peace and stability in Ukraine near our border, it affects us and negatively tells on our partners in Europe and on the global economy as a whole,” he stated, according to Itar-Tass.
The United Nations has also warned European sanctions on Russia could backfire.
The new sanctions largely target Russia’s defense industry, but the legislation also empowers Obama to provide lethal military aid to Ukraine.
Under the legislation, the U.S. will provide a new US$350 million aid package to Kiev. In a major address to the nation on Thursday, President Vladimir Putin will discuss new measures to handle Russia’s economic stress and his government’s efforts to promote peace in Ukraine, according to Peskov.
“Russia is consistently doing all to contribute to the establishment of peace,” Peskov stated.
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