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Last year, cases of tendinitis, bursitis, repetitive strain injuries and back problems caused workers to take a total of 2,018 periods of sick leave, some of which lasted as many as 15 days, according to the prosecutors' statement.
"[The sum of R$250 million] may seem excessive at first, but putting things into perspective, it is equivalent to what the defendant makes as a profit around the world in less than two days," the prosecutors' office said.
(Read more: Apple vs. Samsung debate: Which really makes more money?)
As well as demanding R$250 million in damages, prosecutors are asking Samsung to give employees a 10-minute break every 50 minutes.
Samsung, the world's largest maker of smartphones and televisions, has also come under criticism in China over working conditions at its suppliers.
Last year, New York-based China Labor Watch released reports including allegations of child labor. It also alleged illegal practices at six factories that were wholly or mostly owned by the South Korean group, including working hours in excess of legal limits. Samsung denied allegations of child labor, citing a "zero tolerance" policy.