The Scottish course, which last hosted the British Open in 2013, has been told it won't be considered as a venue for golf's oldest major tournament unless it lets females join.
With 750 Muirfield members eligible to vote, the two-thirds majority needed for a change to take place was not reached, prompting a response from Royal & Ancient (R&A), which organizes the British Open.
"The R&A has considered today's decision with respect to the Open Championship," the R&A said in a statement. "The Open is one of the world's great sporting events and going forward we will not stage the Championship at a venue that does not admit women as members.
"Given the schedule for staging the Open, it would be some years before Muirfield would have been considered to host the Championship again. If the policy at the club should change we would reconsider Muirfield as a venue for the Open in future."
While fellow British Open venues the Royal & Ancient Club of St. Andrews and Royal St. George's have both admitted female members in recent years, the East Lothian course has chosen not to follow suit after a two-year consultation process.
The R&A was heavily criticized for using Muirfield, which has staged 16 British Opens, as its host venue in 2013 due to the club's membership policy. Established in 1744 at Leith -- a port district in Edinburgh -- it moved to its current East Lothian site in 1891.
The decision from Muirfield's members will come as a setback to organizers of the global game, who have come under increasing pressure to abandon what is seen as an archaic single-gender policy.
The R&A abandoned its 260-year male-only membership policy in 2014
, while Royal St. George's followed last year by admitting women for the first time in 128 years.
Muirfield and Royal Troon are the two remaining traditional British Open host venues to be men-only domains.
However, Royal Troon, which will host this year's event and currently has separate men and women's clubs, is consulting members over whether to make a change.
Across the Atlantic, in 2012 Augusta National allowed female members to join the prestigious U.S. club for the first time in 80 years.
Augusta, which hosts the Masters, faced years of protests with President Obama adding pressure on the organization. Former U.S. Secretary of State Condoleezza Rice and South Carolina financier Darla Moore became the first women to be admitted as members.