FIFA says Barcelona will not get special permission to sign a defender after Thomas Vermaelen was sidelined by injury for several months.
Barcelona sought an emergency signing under "exceptional situations" in Spanish federation rules, despite facing a one-year FIFA transfer embargo.
FIFA says it told Barcelona on Monday that its disciplinary committee did not "conceive an exception" for signing players "outside the official periods of registration."
FIFA's ban was imposed because Barcelona broke rules on international signings of young players. It takes effect in January, pending an appeal verdict this month from the Court of Arbitration for Sport.
Barcelona signed Vermaelen in the off-season when FIFA froze the ban until appeals were completed.
The Belgium international has not played since his reported 19 million euro ($25.4 million) move from Arsenal.
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