The White House decried alleged human rights violations and arrests carried out in response to anti-government protests. It called on the South American nation to release political prisoners, including students and opposition leaders.
Venezuela will respond to the measures "soon," the country's foreign minister told Reuters.
Venezuela has dealt with increasing prices and limited access to basic goods. Inflation increased by 64.3 percent last year, and is seen staying above 60 percent this year.
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Low crude prices have hit Venezuela, a major oil producer, and the country risks defaulting on its sovereign debt this year.
Several critics of President Nicolas Maduro, including Caracas Mayor Antonio Ledezma, have been imprisoned in recent months. Ledezma was arrested in February for his purported involvement in an alleged coup.
Obama's order targets officials who have allegedly acted to restrict free expression or other "democratic processes" or have violated human rights in response to anti-government protests. Individuals named included security, intelligence and police officials.
"We are committed to advancing respect for human rights, safeguarding democratic institutions, and protecting the U.S. financial system from the illicit financial flows from public corruption in Venezuela," the White House said.
The White House stressed that the sanctions would not target "the people or economy of Venezuela." The order would not affect the Venezuelan energy sector, Reuters reported, citing a senior administration official.
It expands on legislation passed last year in which the U.S. pledged to support democratic processes and hold Venezuelan officials accountable for human rights violations.