The Barcelona midfielder, who scored two international goals and who won the World Cup with Spain in 2010 and the European Championship two years later, wrote on Instagram: "I would like to announce that after almost 15 years and 143 matches, the moment has arrived to bid farewell to the national team."
Busquets made his debut for La Roja against Turkey in April 2009 and went on to make 143 appearances, behind only Iker Casillas and Sergio Ramos in the country's history.
The 34-year-old captained Spain in each of their four games at the World Cup in Qatar, missing a penalty in the shootout as Luis Enrique's side were knocked out by Morocco at the last-16 stage.
In the Instagram post, Busquets paid tribute to the variety of people involved in his successful Spain career.
"I would like to thank all the people who have accompanied me on this long road," he wrote.
"From Vicente del Bosque who gave me the opportunity to Luis Enrique for making me enjoy until the last second.
"I also thank Julen Lopetegui, Fernando Hierro and Robert Moreno for their trust, as well as all their staff.
"And, of course, to each and every one of my team-mates, with whom I have struggled to try to take the team where it deserved, with more or less success but always giving everything and with the greatest of pride.
"It has been an honour to represent my country and take it to the top.
"And, of course, to each and every one of my team-mates, with whom I have struggled to try to take the team where it deserved, with more or less success but always giving everything and with the greatest of pride.
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