Colombian coach Jose Pekerman called up midfielder Carlos Carbonero to replace the injured Aldo Ramirez in his World Cup finals squad.
Carbonero, 23 and who recently won the Argentinian league title with River Plate, has been capped just once and did not feature in Pekerman's original provisional 30-man squad for the finals in Brazil.
Ramirez, 33, had been struggling to be fit after suffering a knee injury in April and his withdrawal is a further blow to Colombia's hopes of at least progressing from their group.
Talismanic striker Radamel Falcao was ruled out last week after failing to recover from a serious knee injury.
Colombia have been drawn in Group C along with goalshy Greece, the talented but ageing Ivory Coast and Japan.
The South Americans open their campaign against Greece on June 14.