Chile need a win in the hot and humid Caribbean port city to secure a ticket to their second finals in succession with one qualifier to spare.
Colombia could also qualify for the 2014 World Cup in Brazil even if they lose to Chile if the match between Ecuador and Uruguay, taking place at the same time (2100 GMT), ends in anything but a draw.
Up in the Andes at the Atahualpa in Quito, 2010 semi-finalists Uruguay visit Ecuador in a clash of teams separated only by goal difference.
Argentina, at home to Peru in Buenos Aires (2300), have already qualified, while Venezuela host Paraguay at Puerto La Cruz (2030) hoping a win will keep alive their slim hopes of reaching the finals for the first time.
The top four in the South American group go through to the next year's World Cup with the fifth-placed team after the final round on Tuesday qualifying for a playoff against Jordan.
With two rounds to go, second-placed Colombia have 26 points, two ahead of Chile and four in front of Ecuador and Uruguay, with sixth-placed Venezuela three points further back and having played one match more.
The climate in Barranquilla could be a handicap for the Chileans used to cooler temperatures but coach Jorge Sampaoli said his main worry was opponents on a run of five successive home wins.
"Our opponents, with the arrival of their present trainer, have marked a difference with the rest of the national teams." Sampaoli said, referring to fellow Argentine Jose Pekerman.
"They've scored a lot of goals at home (14), had none scored against them and won all their matches (at home since the new coach took over)," he told reporters.
Pekerman, who steered Argentina to the 2006 quarter-finals in Germany, took charge after Colombia lost 2-1 to Argentina in Barranquilla two years ago.
Uruguay, on a run of three wins after beating Colombia 2-0 in Montevideo last month, want to avoid having to try qualifying via the playoff for the third World Cup in row.
"With a point we can secure the playoff (at least) which for us three matches ago would have been a miracle," said striker Luis Suarez, looking back on a mid-series slump when they took only two points from six matches.
"Now we know we have good chances of qualifying directly which is what we want, to avoid more suffering," added Suarez, who with 10 goals is the group's joint top scorer with Argentina's Lionel Messi.
Uruguay have featured in playoffs for the last three World Cups, beating Australia to reach the 2002 finals, losing to the same opponents for the 2006 tournament and overcoming Costa Rica to get to South Africa in 2010.
"There is one more match after this one in Quito, but the importance of that match against Argentina will depend on the result we bring home from Ecuador," Suarez said in Montevideo.
Ecuador, who rely mainly on Felipe Caicedo for their goals after fellow striker Cristian Benitez died suddenly of heart failure in July, are looking to return to the finals after missing out on South Africa.