The International Atomic Energy Agency had warned Saturday that the "disproportionate and unprecedented" move would seriously hamper its work.
Iranian media and a Western diplomat said the decision concerned eight inspectors, all from France and Germany.
Brussels, which acts as coordinator for the 2015 nuclear deal between Tehran and world powers known as the JCPOA, said it was "highly concerned" by the development.
"Particularly worrying is the direct and severe impact of this decision on the agency's ability to conduct its verification activities, which includes the monitoring of the Joint Comprehensive Plan of Action [JCPOA]," a spokesman said.
"The EU urges Iran to reconsider its decision without delay."
Iran's foreign ministry said Saturday that the move was in retaliation for "political abuses" by the United States, France, Germany and Britain.
In 2015, major world powers reached the accord with Iran under which Tehran would curb its nuclear programme in exchange for relief from crippling economic sanctions.
But that started to unravel in 2018 when Donald Trump, the US president at the time, withdrew from the deal and reimposed sanctions.
Tehran in turn stepped up its nuclear programme, while continuing to deny that it harbours ambitions of developing nuclear weapons capability.
Efforts to revive the deal have been fruitless so far.
Britain, France and Germany, European signatories to the deal, announced last week that they would not lift sanctions on Iran next month as originally foreseen.
Iran warns EU sanctions are 'illegal'
Tehran (AFP) Sept 15, 2023 -
Iran has warned the European Union that its decision to maintain sanctions on the Islamic republic beyond a deadline set in a landmark nuclear deal was "illegal".
Britain, France, and Germany said that "EU and UK sanctions will be maintained," according to a statement from the British foreign ministry released on Thursday.
The series of British, EU and US sanctions "on individuals and entities involved in Iran's missile, nuclear, and other weapons programmes" were supposed to be lifted on October 18, the ministry said.
The deal agreed with world powers in 2015 was aimed at curbing Iran's nuclear programme in exchange for the lifting of international sanctions.
But the accord began unravelling in 2018 when then US president Donald Trump unilaterally withdrew from it and began reimposing sanctions and Iran retaliated by dropping some of its obligations under the agreement.
Iran's foreign ministry said it considers "the decision of the EU and three European countries as an illegal act with malicious intentions, and contrary to their obligations under the nuclear agreement".
"We warn the European parties against taking actions that will certainly complicate the management of their relations and have a negative impact on... negotiations for the lifting of sanctions," it said in a statement issued late Thursday.
Efforts to revive the deal have been fruitless so far with European-led talks on hold since 2022.
In its statement, the British foreign ministry said the decision to keep the sanctions in place had been based on "continued Iranian non-compliance with their... commitments and ongoing nuclear escalation".
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