Colombia's star striker Radamel Falcao has lost his battle to be fit for the World Cup.
National coach Jose Pekerman named his tournament squad on Monday evening during a press conference held in Buenos Aires, Argentina.
Falcao, who suffered a serious knee injury when on duty for his club side Monaco in January, was left off the 23-man list, with Pekerman deciding he would not be ready.
The former Atletico Madrid frontman, who was a big-money signing by Monaco last summer, looked to have no hope of making his country's squad for the tournament in Brazil when he sustained the injury, however his recovery then progressed swiftly.
Pekerman said last week he would "wait for Falcao until the last day, the last minute" before deciding on his fitness, after consulting with medical experts, and the final decision went against the 28-year-old.
"(Luis Fernando) Muriel, (Amaranto) Perea and Falcao are not going to be at the World Cup, Jose Pekerman has announced," said a message on the national team's official Twitter account.
Colombia face Greece, Ivory Coast and Japan at Brazil 2014.
Falcao said following the announcement, according to Colombian newspaper El Pais: "I was very excited but now I'll have to support the team from afar and I wish them well, hoping they play a good World Cup."
After taking the tough decision, Pekerman called it "the saddest day I've had since becoming Colombia coach".
Colombia World Cup squad: David Ospina (Nice), Faryd Mondragon (Deportivo Cali), Camilo Vargas (Santa Fe); Camilo Zuniga (Napoli), Pablo Armero (Napoli), Cristian Zapata (AC Milan), Mario Yepes (Atalanta), Carlos Valdes (Philadelphia Union - on loan to San Lorenzo), Santiago Arias (PSV Eindhoven), Eder Alvarez-Balanta (River Plate); James Rodriguez (Monaco), Abel Aguilar (Toulouse), Carlos Sanchez (Elche), Fredy Guarin (Inter Milan), Juan Fernando Quintero (Porto), Aldo Ramirez (Morelia), Alexander Mejia (Atletico Nacional), Victor Ibarbo (Cagliari), Juan Guillermo Cuadrado (Fiorentina); Jackson Martinez (Porto), Teofilo Gutierrez (River Plate), Carlos Bacca (Sevilla), Adrian Ramos (Borussia Dortmund)