Yesterday was the last day of publication for the free daily newspaper, which was published in seven Spanish cities, including Valencia, Alicante, Castellon de la Plana and Albacete.
Metro's Spanish edition launched in 2001 and by 2004 had achieved profitability.
Prior to its closure, Metro claims it was the fifth most read daily newspaper in Spain with more than 1.8m daily readers after paid for titles such as El Mundo and El Pais.
In a statement Metro International, said: "With increased competition and a steadily declining advertising market the operation has been experiencing losses during the last years.
"Several merger opportunities have been investigated during 2008, however, the size of the combined losses of the Spanish free press market made it extremely difficult to find an alternative that would have improved the future financial performance of Metro Spain."
The company said it invested €25.5m (£23.1m) in Metro Spain up until it closed yesterday.
The closure is expected to give rise to a book loss of between €5 to €6m, which includes non-cash items.
Per Mikael Jensen, chief executive and president of Metro International, said: "Despite dedicated efforts from our Spanish management team and staff, it is with deep regret that we have taken the difficult decision to close down our operations in Spain.
"Even though Metro in Spain has been losing less money than its Spanish free competitors, the worsening Spanish economic downturn, which during the beginning of 2009 has resulted in a collapsing advertising market, has now resulted in unsustainable losses.
"The stiff competition coupled with a forecasted continued weakening advertising market for Spain, makes the closure of our operations the only rational decision at this time.
"A continued investment in Spain cannot be justified at this point and we are therefore focusing our resources on growth areas where we can create long term shareholder value."