The EU has progressively clamped down on animal testing since the 1990s and banned most such products in 2009, but it left a few exemptions for several toxicity tests which will now cease.
The ban applies to all products, wherever in the world they come from.
The European Commission has “thoroughly assessed the impacts of the marketing ban and considers that there are overriding reasons to implement it,” a statement said.
“This is in line with what many European citizens believe firmly: that the development of cosmetics does not warrant animal testing.”
EU Health Commissioner Tonia Borg said Brussels would continue “supporting the development of alternative methods and to engage with third countries to follow our European approach”.
Earlier this month, Japanese cosmetics giant Shiseido said it was dropping animal-tested products, with some exceptions where such tests were the only way of proving the safety of goods already on sale.
“Our business partners that supply material to us will not rely on animal testing while we will no longer outsource such testing to outside labs,” Shiseido said.
Activists have for years pressured cosmetic firms and other companies that use animal testing to find alternatives to the practice, which they say is cruel and unnecessary.
Shiseido, which dropped animal testing at its own labs in 2011, said it could ensure the safety of its products through other means, including using data from past experiments, human volunteers and other kinds of testing.