The Shakhtar Donetsk striker played 21 minutes during Croatia's 2014 World Cup campaign, coming on as a substitute in their 4-0 win over Cameroon in Brazil.
And it would be the 31-year-old's final appearance for Croatia, who sits second in the nation's all-time international scorers list behind Davor Suker (45 goals).
"I was selected 64 times in the last 10 years, from my first match for Croatia against Ireland in 2004. I scored 29 goals, which makes me the most prolific active striker in the team," Eduardo said.
"With great pride and joy I play in the Croatian shirt and presented Croatian football around the world. It was important to justify the trust that the fans gave me for which I am especially grateful.
"I'm in the team among players, coaches and many other people (medical staff, technical staff, and others) who are very much, everyone in their own way, important and I made many friends.
"I also thank all the coaches who had faith in me. I will remember playing for Croatia only in a nice way."
Coach Niko Kovac paid tribute to the Brazil-born forward's national service, which began in November 2004.
"Thanks, Eduardo. I wish him all the luck in the world for the rest of his career," Kovac said.
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