The former Galatasaray goalkeeper, now 43, became the oldest player to take part in a World Cup finals match when he replaced David Ospina in the group-stage win over Japan.
He had earlier announced that this summer's tournament in Brazil would be his last as a player and the Deportivo Cali icon has now confirmed his career has reached its conclusion.
"This is my last stadium as a professional player," he said on his official Instagram page. "I'm proud to be part of this wonderful group."
Mondragon, who began his career with Cali, also enjoyed spells with Independiente and Santa Fe before moving to Europe to join Real Sociedad in 1999.
His most successful period came with Turkish giants Galatasaray, where he won two Super Lig titles and a domestic cup between 2001 and 2007, after which he joined Koln.
Mondragon, who earned a total of 56 caps for Colombia over a remarkable 23-year career, spent a season with Philadelphia Union in 2011 before returning to Deportivo Cali for two years prior to his retirement.