The former Barcelona captain is in his third season with Inter Miami.
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Now 38, Busquets stated on social media: "I feel like the time has come to say goodbye to my career as a professional footballer. It's been almost 20 years of enjoying this incredible story I always dreamed of.
"I'm retiring very happy, proud, fulfilled, and above all, grateful. Thank you all very much, see you soon."
With Barca, the midfielder won 32 major honours in 18 years, including nine La Liga titles and three Champions Leagues.
He made 143 appearances for Spain and was part of the side that won the 2010 World Cup and 2012 European Championship.