"Despite the positive evolution of the grade one sprain to his left ankle he will be absent from tonight's game," read a statement from the club.
Neymar was only given the all-clear by medical staff at the Camp Nou on Monday after recovering from fracturing a vertebra playing for his country in the World Cup quarter-final against Colombia in early July.
He looked lively in Barca's final friendly match against Mexican outfit Leon on Monday in which they won 6-0 and he bagged a brace.
Barca are looking to make a new start under coach Luis Enrique after they failed to win a major trophy last season.
(Editing by Martyn Herman)
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