Backup Colombia goalkeeper Yineth Varon failed a drug test at the women's World Cup and was provisionally suspended by FIFA on Tuesday.
Varon failed an out-of-competition doping control on Saturday in Leverkusen, Germany, the world governing body of football said.
Colombia played Sweden in its opener on Tuesday in Leverkusen with Sandra Sepulveda in goal, losing 1-0.
Colombia coach Ricardo Rozo wouldn't comment on Varon's suspension after the game.
FIFA did not say what substance was involved, but that the player had the right for her "B" sample to be tested. FIFA medical chief Michel D'Hooghe later declined to comment.
The last doping case at the men's World Cup was Diego Maradona, who was kicked out from the 1994 World Cup in the United States after testing positive for stimulants. He was banned for 15 months.
Last month, five Mexico players tested positive for clenbuterol in pre-competition testing at the Gold Cup. Football officials allowed Mexico to replace the players.
FIFA president Sepp Blatter said on Saturday that contaminated meat was the cause.