The "Like A Prayer" singer was due to arrive on stage at 8pm to begin her show at Brisbane Entertainment Centre, however concertgoers were told that she would not be on stage for another three hours, reported Guardian online.
Fans inside the venue grew restless while they waited for the 57-year-old star, with some threatening to give up waiting for their idol and go home.
One Twitter user wrote, "Madonna you're late. #getonstage #RebelHeartTour #wearegoinghome".
Another added,"Still waiting for Madonna 2 hours late whose (sic) gotta catch the train."
The gig, the first of two dates in Brisbane on her Rebel Heart world tour, is just the latest in a long line of late appearances by the pop superstar.
Earlier this month, she annoyed concertgoers in Auckland, New Zealand by taking to the stage one-and-a-half hours late, while fans in the Philippines were also kept waiting for three hours.