Defender Thomas Vermaelen and midfielder Andre Gomes will miss the Joan Gamper Trophy match on Monday between Barcelona and Chapecoense.
The Blaugrana confirmed their absence in a statement on Monday morning after the players had trained at the Ciutat Esportiva in preparation for the game.
Barça said: "Andre Gomes and Thomas Vermaelen will both be sitting out the game against Chapecoense.
"The Portuguese player took a knock in the hip playing against Gimnástic Tarragona last Friday, while the Belgian hurt his leg in the same match."
Vermaelen is one of the players included in 'Operation Exit' this summer and has offers from England, Italy and Belgium.