President Donald Trump has been a critic of the majority-black city, calling it a 'rat and rodent infested mess'. At one point, Mrs. Trump interrupted her remarks to say, "Hello, everyone", before continuing.
The boos began as she greeted the more than 2,500 attendees of the B'More Youth Summit, held at the University of Maryland's Baltimore campus.
She was introduced as "somebody who has made it their personal mission to use their platform to make a difference in this particular area, with wellness and addiction and particularly the opioid crisis that we're living through right now".
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First ladies are usually spared the rougher edges of U.S. politics and Melania Trump has been no exception, despite the bruising nature of her husband's presidency.
During her speech, the first lady called on leaders to educate themselves on the opioid epidemic and to help educate the youth about the benefits of a drug-free lifestyle.
She smiled and waved to the crowd despite the boos.
And she tells the students: "I am in this fight with you and I am fighting for you". As she left the stage, she again met with a mixture of cheers and boos.