In her dazzling career, Makeba performed with musical legends from around the world - jazz maestros Nina Simone and Dizzy Gillespie, Harry Belafonte, Paul Simon - and sang for world leaders such as John F. Kennedy and Nelson Mandela.
"Her haunting melodies gave voice to the pain of exile and dislocation which she felt for 31 long years. At the same time, her music inspired a powerful sense of hope in all of us," Mandela said in a statement.
He said it was "fitting" that her last moments were spent on stage.
The Pineta Grande clinic in Castel Volturno, near the southern city of Naples, said Makeba died early Monday of a heart attack.
Makeba collapsed on stage Sunday night after singing one of her most famous hits "Pata Pata," her family said in a statement. Her grandson, Nelson Lumumba Lee, was with her as well as her longtime friend, Italian promoter Roberto Meglioli.
"Whilst this great lady was alive she would say: 'I will sing until the last day of my life'," the statement said.
Castel Volturno Mayor Francesco Nuzzo said Makeba sang at a concert in solidarity with six immigrants from Ghana who were shot to death in September in the town, an attack that investigators have blamed on organized crime.
The death of "Mama Africa," as she was known, plunged South Africa into shock and mourning.
"One of the greatest songstresses of our time has ceased to sing," Foreign Affairs minister Nkosazana Dlamini Zuma said in a statement.
"Throughout her life, Mama Makeba communicated a positive message to the world about the struggle of the people of South Africa and the certainty of victory over the dark forces of apartheid and colonialism through the art of song."
Makeba wrote in her 1987 memoirs that friends and relatives who first encouraged her to perform compared her voice to that of a nightingale. With her distinctive style combining jazz with folk with South African township rhythms, she was often called "The Empress of African Song."
The first African woman to win a Grammy award, Makeba started singing in Sophiatown, a cosmopolitan neighborhood of Johannesburg that was a cultural hotspot in the 1950s before its black residents were forcibly removed by the apartheid government.
She then teamed up with South African jazz trumpeter Hugh Masekela - later her first husband - and her rise to international prominence started when she starred in the anti-apartheid documentary "Come Back, Africa" in 1959.
When she tried to fly home for her mother's funeral the following year, she discovered her passport had been revoked. It was 30 years before she was allowed to return.
In 1963, Makeba appeared before the U.N. Special Committee on Apartheid to call for an international boycott of South Africa. The South African government responded by banning her records, including hits like "Pata Pata," "The Click Song" ("Qongqothwane" in Xhosa), and "Malaika."
Makeba received the Grammy Award for Best Folk Recording in 1966 together with Belafonte for "An Evening With Belafonte/Makeba." The album dealt with the political plight of black South Africans under apartheid.
Thanks to her close relationship with Belafonte, she received star status in the United States and performed for President Kennedy at his birthday party in 1962. But she fell briefly out of favor when she married black power activist Stokely Carmichael - later known as Kwame Ture - and moved to Guinea in the late 1960s.
Besides working with Simone and Gillespie, she also appeared with Paul Simon at his "Graceland" concert in Zimbabwe in 1987.
After three decades abroad, Makeba was invited back to South Africa by Mandela, the anti-apartheid icon, shortly after his release from prison in 1990 as white racist rule crumbled.
"It was like a revival," she said about going home. "My music having been banned for so long, that people still felt the same way about me was too much for me. I just went home and I cried."
Makeba courted controversy by lending support to dictators such as Togo's Gnassingbe Eyadema and Felix Houphouet-Boigny from Ivory Coast, performing at political campaigns for the veteran leaders even as they were violently suppressing the movements for democracy that swept West Africa in the early 90s.
The first person to give her refuge was Guinea's former President Ahmed Sekou Toure who was accused in the slaughtering of 10 percent of the population.
Makeba, though, insisted that her songs were not deliberately political.
"I'm not a political singer," she insisted in an interview with Britain's Guardian newspaper earlier this year. "I don't know what the word means. People think I consciously decided to tell the world what was happening in South Africa. No! I was singing about my life, and in South Africa we always sang about what was happening to us - especially the things that hurt us."
Makeba announced her retirement three years ago, but despite a series of farewell concerts she never stopped performing. When she turned 75 last year, she said she would sing for as long as possible.
Makeba is survived by her grandchildren, Nelson Lumumba Lee and Zenzi Monique Lee, and her great-grandchildren Lindelani, Ayanda and Kwame.