Pato, who came close to joining Paris Saint-Germain last week before committing his future to the Serie A champions, picked up the muscle strain during Milan's 2-1 extra-time victory against Novara in the Coppa Italia round of 16 on Wednesday.
The 22-year-old struck the winning goal at San Siro, but ended the match struggling with an injury.
Milan's medical staff have since examined the former Internacional man and expect him to be unavailable for the next month.
"AC Milan can announce today that Pato has been subjected to tests and after specialist medical advise, he was diagnosed with an injury the biceps femoris of the left thigh," the club said in a statement.
"The prognosis is three to four weeks (out), unless there are complications."
Milan meet Novara again in an away match on Sunday as they look to recover from a 1-0 derby defeat to Inter last weekend.
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