Brazil's 2-0 friendly win over Asian champions Qatar in Brasilia on Wednesday came at a heavy cost after they lost striker Neymar for this month's Copa America because of an ankle ligament injury.
Neymar limped off in the 17th minute after he appeared to twist his right ankle and the Brazilian Football Confederation (CBF) later confirmed he would miss the tournament.
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"Neymar was evaluated and underwent scans that confirmed a ligament rupture in the ankle," CBF said in a statement.
"Due to the seriousness of the injury, Neymar will not recover in time to participate in Copa America in Brazil."
The CBF said they would meet on Thursday to pick a replacement for Neymar in the squad for the tournament, which kicks off on June 14.
Brazil were already leading 1-0 courtesy of Richarlison's 16th-minute header when Neymar departed and Gabriel Jesus added the second eight minutes later from close range.
The five-time world champions were rarely troubled by a limited Qatar side, who missed a VAR-assisted penalty in injury time.
"We won and the feeling is that we won well," said man of the match Philippe Coutinho. "We pressured them and we did what we wanted to do."
Brazil will face Bolivia in their Copa America opener on June 14 in Sao Paulo without their best player.