The former England captain had been included on coach Stuart Pearce's 35-man shortlist, and was expected to be named in the final squad of 18 as one of three over-age players.
Craig Bellamy had already confirmed his place in Pearce's plans and fellow Welshman Ryan Giggs is also reportedly set to be involved.
Liverpool midfielder Joe Cole and Manchester City's Micah Richards have also been linked after both missed out on a place in England's squad for Euro 2012.
"Everyone knows how much playing for my country has always meant to me," Beckham said in a statement. "So I would have been honored to have been part of this unique Team GB squad.
"Naturally I am very disappointed, but there will be no bigger supporter of the team than me. And like everyone, I will be hoping they can win the gold."
Beckham had hoped to make a comeback on the international stage having captained England for nearly six years between 2000 and 2006. He is the Three Lions' most-capped outfield player with 115 national team appearances to his name.
Now 37, the former Manchester United star last turned out for his country in 2009 having left Real Madrid two years earlier to play in MLS.
Goal.com has a new podcast! Goal Daily Podcast is a MUST for all Soccer fans and discusses the very latest news and views. What’s more when you sign up the service is FREE for the first month and you can cancel anytime.
Follow GOAL.COM USA on