The National Assembly called for the head of state to "declare persona non grata the head of the diplomatic delegation" from the EU, Isabel Brilhante Pedrosa, in order to proceed with her expulsion.
"I vote with both hands for the European Union representative to be declared persona non grata," said Assembly Speaker Jorge Rodriguez.
Pedrosa had been declared persona non grata once before, when the EU imposed a previous round of sanctions on Venezuelan officials in July last year. Back then, Caracas gave her 72 hours to leave the country, but the government later backed down.
The National Assembly also called on Tuesday for a revision of the agreement on the EU's presence in Caracas.
The vote came after the EU foreign ministers agreed on Monday to impose sanctions on 19 Venezuelan officials for "undermining democracy."
Two members of the assembly were among the targeted officials. The governor of Zulia State, the commander of the armed forces, and three members of the Electoral Council, including its president, were also added to the EU's list of the individuals targeted with sanctions.
That would bring to 55 the total number of the members of the Venezuelan government targeted with asset freezes and travel bans by the EU.
The EU expanded its list of sanctions against Venezuelan officials after it refused to recognize the results of the December 2020 legislative elections, which saw Maduro win total control of parliament.
The United States and several Latin American countries have also rejected the election results.
They have long been supporting opposition figure Juan Guaido, who has plans to maintain a parallel parliament of shadow opposition lawmakers at an assembly that has already been declared defunct by Caracas.