The Darts Regulation Authority (DRA) said its new eligibility policy followed a review by Dr Emma Hilton, an academic developmental biologist who has published several papers on sex and sporting categories.
"As a result of its review, the DRA is satisfied that to achieve fair competition in darts, only biological females should be eligible to compete in women’s tournaments regulated by DRA rules," the authority said in a statement.
"The DRA seeks to be inclusive and all players - irrespective of their biological sex, legal sex, and/or gender identity - may compete in open tournaments and are encouraged to do so."
According to Hilton's report, darts is a "gender-affected sport", with "multiple, small-magnitude sex differences" accumulating to generate a male advantage over female players.
The new regulations come weeks after the International Olympic Committee (IOC) announced that only biological female athletes, whose gender has been determined by a one-time gene-screening test, will be eligible to compete in female category events at the Olympic Games.
Noa-Lynn van Leuven, a Dutch transgender player who has won several women's tournaments, said the ruling would force her into retirement at 29.
"The DRA just decided that trans women are no longer allowed in women's events, which basically means I'm out," she said in an Instagram post.
"This is another huge hit for the trans community, especially after recent decisions made by the IOC. Every day it's getting harder and harder for trans people just to exist, to compete."