During a engagement to Lichfield Cathedral on Monday, King Charles was confronted by a man who questioned him about his brother's connections with disgraced financier Jeffrey Epstein.
The man demanded to know how long the King had been aware about his brother's relationship with Jeffrey Epstein, and also questioned whether he had instructed the police to "conceal" for Andrew.
Despite the protest, there were crowds of fans and applause for the King, who was in Staffordshire for an ceremony.
Prince Andrew's associations with Epstein have been under intense scrutiny in recent weeks. The prince, who relinquished his titles earlier this month, has strongly denied any wrongdoing.
"How long have you known about Prince Andrew and Epstein? Have you asked the police to cover up for him?"
The monarch ignored the questions but continued greeting fans, some of whom chanted "God Save The King". One supporter can be heard asking him for a hug.
Another woman outside the venue was also observed telling the heckler to "go away" and to "shut up".
The scandal over Prince Andrew's links to Jeffrey Epstein has been revived by sexual abuse allegations repeated in Ms. Giuffre's posthumously-published book.
In the publication, Ms Giuffre says she had sexual encounters with the royal on multiple times, including once with Jeffrey Epstein and "several other minors"
Prince Andrew, who reached a out-of-court agreement with Ms Giuffre in recent years, has consistently denied any wrongdoing. In a previous interview, he told a news program he did not remember meeting her "ever", and that they "did not have any sort of sexual contact."
But in recent times, new inquiries have been raised about how he is able to finance his way of life despite not being a active member of the royal family.
The revelation that he has only ever paid a token annual rent on his Royal Lodge has intensified the scrutiny over his living arrangements - even though he made large payments up front, including for renovations.
The political party said he should be called in front of members of parliament to give testimony about his rental agreement of his lodge.
But last week, the government refused to give MPs time to debate the prince's titles or his residence, despite increasing calls for scrutiny.
The royal household has not issued a statement on whether the Duke of York might move out of his current residence - or where he might go.
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