The Financial Conduct Authority (FCA) and the Prudential Regulation Authority (PRA) have fined Staley £642,430, which is 10 per cent of his annual income, saying he "failed to act with due skill, care and diligence in the way he acted in response to an anonymous letter".
The two organisations also announced "special requirements" for whistleblowing at Barclays, meaning the bank must now report annually to the regulators detailing how it handles whistleblowing, and provide "personal attestations" from the senior managers responsible.
"Mr Staley breached the standard of care required and expected of a chief executive in a way that risked undermining confidence in Barclays whistleblowing procedures," said the FCA's Mark Steward.
"Whistleblowers play a vital role in exposing poor practice and misconduct in the financial services sector. It is critical that individuals are able to speak up anonymously and without fear of retaliation if they want to raise concerns."
Staley was widely condemned as he attempted to identify the writer of an anonymous letter, claiming to be from a Barclays shareholder, which he received in 2016.
The letter contained various allegations about the bank, some of which concerned Staley, and the FCA and PRA said this should have prompted the chief executive to take a step back.
However, the regulators said last month that they were "not alleging that [Staley] acted with a lack of integrity or that he lacks fitness and propriety to continue to perform his role as group chief executive officer".