The Barcelona court rejected the Brazilian's petition because of a flight risk despite Alves' lawyers stating their client would accept any court-imposed measures, such as turning in his passports and wearing a tracking device, if freed on bail.
"We consider that no other precautionary measure can neutralise that risk with sufficient guarantees and his flight would prevent the holding of the trial that requires the presence of the individual being investigated," the Barcelona court stated on Monday.
Alves had been denied a similar request in February and May, for that reason.
The Brazilian is currently in pre-trial detention until a date is set for a hearing.
Alves, 40, was provisionally detained in January after being accused of sexually assaulting a woman at a nightclub in Barcelona on Dec. 30.
He has denied wrongdoing and said that sex with the woman was consensual.
A judge ordered him to be jailed without bail after analysing the initial probe by authorities and hearing testimony from Alves, the alleged victim and witnesses.
Alves changed his version of events four times; the victim's statement has remained the same and biological test results found traces of Alves' semen in the victim's body.
Under Spain's sexual consent law passed last year, sexual assault covers a wide array of crimes from online abuse and groping to rape, each with different possible punishments.
A case of rape can receive a maximum sentence of 15 years.
Alves won 42 football titles, including three Champions Leagues with Barcelona and two Copa Americas with Brazil.
He played in his third World Cup last year in Qatar.
Mexican club Pumas terminated Alves' contract after the player was accused of sexual assault.