The Colombia shot-stopper entered the pitch in the 85th minute, replacing David Ospina to a rapturous applause from los Cafeteros' fans during the 4-1 Group C win.
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Mondragon not only broke Milla's record, but also entered into the annals of World Cup history by becoming the player with the most time elapsed between his most recent appearances, having last featured 16 years ago at France '98.
This record was formally held by the Switzerland's Alfred Bickel, who experienced a 12-year gap between playing in World Cups which, interestingly, were also held in France and Brazil in 1938 and 1950 respectively.
Mondragon, who has 57 international caps, is set to retire from football after the tournament in Brazil, bringing to an end a 24-year career.
The goalkeeper left Deportivo Cali at the end of the previous campaign after spending time in eight countries, earning two league titles with Galatasaray and a Copa Libertadores with Independiente.