"I am not a candidate to the presidency of the Eurogroup," he told reporters, without giving further details.
His comments came despite Spanish Prime Minister Mariano Rajoy backing de Guindos as potential head of the group of eurozone finance ministers in an interview published Sunday in several newspapers.
De Guindos was the main challenger to replace Dijsselbloem as the head of the Eurogroup when his first term expired in 2015 but in the end the Dutchman secured his re-election to the powerful post.
But he has run into controversy since he said in an interview last week that southern European countries blew their money on "drinks and women".
Southern Europeans reacted strongly, with Portugal's prime minister and former Italian premier Matteo Renzi calling for Dijsselbloem, who is also Dutch finance minister, to step down.
His job was already up in the air after his party lost out in elections in the Netherlands earlier this month. His mandate as head of the Eurogroup lasts until January 2018.
De Guindos said Dijsselbloem's comments were "regrettable," and pointed out that Spaniards are "very under-represented" in executive positions in European institutions.