BlackRock's iShares Bitcoin Trust (IBIT) recorded a net outflow of approximately $527.8 million on Wednesday.
This represents the fund's second-largest single-day withdrawal on record.
The redemption from market-leader IBIT coincided with a decline in Bitcoin's price.
U.S. spot Bitcoin ETFs logged eight consecutive days of cumulative net outflows.
Total outflows exceeded $2 billion for the month of May.
These sustained outflows suggest cooling institutional demand and cautious market sentiment.
Analysts attribute the shift to macroeconomic factors and profit-taking.