The pair discussed economic opportunities and preventing Venezuela becoming a place for "America's adversaries", a US official said.
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The two-hour meeting was aimed at building trust and communication following the US seizure of Venezuela's former president Nicolás Maduro almost two weeks ago.
Ratcliffe and Rodríguez discussed potential economic collaboration and that "Venezuela can no longer be a safe haven for America's adversaries", according to the US official.
The meeting took place on the same day Rodríguez gave her first state of the union address since taking over as interim president, where she announced oil industry reforms to allow more foreign investment - a move away from Maduro's policies.
During her speech to the nation, Rodríguez said she was not afraid to face the US "diplomatically through political dialogue," adding Venezuela had to defend its "dignity and honour".
President Donald Trump has said US oil companies will move into Venezuela and make money that will go to people there and to the US, with a top official saying the US would control sales of sanctioned Venezuelan oil "indefinitely".
Trump has asked oil companies to invest at least $100bn (£75bn) in Venezuela, but one executive said last week the country is currently "uninvestable".
Rodríguez, the former vice-president, was sworn in on 5 January after US forces seized Maduro and his wife Cilia Flores in an operation in Caracas. They are now detained in New York, where they have pleaded not guilty to drug trafficking and other charges.
The US official described the meeting between Rodríguez and Ratcliffe as "historic", adding that Ratcliffe was the first cabinet-level official to travel to Venezuela.
The meeting also happened on the same day that Trump met opposition leader Maria Corina Machado in Washington, who presented the US president with her Nobel Peace Prize winning medal.
In her state of the union address, Rodríguez told Venezuelans it was "very difficult" to deliver Maduro