If they win and Independiente del Valle fail to beat Atletico Mineiro, that will ensure qualification with a game to go.
Match preview
It has been a contrast of fortune for these sides in this season's Copa Libertadores, with Tolima in a good position to qualify, whereas America Mineiro have just one point from four games.
Mineiro could still qualify for the knockout stage, but their form and far inferior goal difference makes that unlikely, making a third-placed finish and dropping into the Copa Sudamericana their best hope.
Tolima, meanwhile, have excelled in their first Copa Libertadores campaign in nine years and are unbeaten in their past three continental outings.
This has left them just one point off last year's semi-finalists Atletico Mineiro, but with a daunting away trip to the Brazilian side next week, it is important that they do not slip up here.
However, their recent form in all competitions suggests that they should be fine having avoided defeat in their last eight games, which also has the Colombian side flying high in second in the league.
Their form has largely stemmed from a high scoring attack and it is a forward line that will be going up against a team, who have the fourth worst defensive record in this season's Copa Libertadores.
Wednesday is a must-win for Mineiro though, because anything other than three points will leave them with no hope of qualifying for the knockout stage, or the Copa Sudamericana.
They have been put into a very tough group against sides who have just proved to be too strong for them, and that has shown through their poor defence.
Mineiro have conceded eight goals from the opening four rounds, including three in this game's reverse fixture which ended up being a 3-2 away win for Tolima.
However, with this fixture now being a home one for Hernan Torres's side, who have qualification very much in their sight, it is hard to see a scenario in which America Mineiro come out on top.
If Tolima do take all three points, then that could set up a winner-takes-all match against Atletico Mineiro next week, in which the winner finishes top of the group.
Deportes Tolima Copa Libertadores form:
Deportes Tolima form (all competitions):
America Mineiro Copa Libertadores form:
America Mineiro form (all competitions):
Team News
Following their 1-1 draw over the weekend, Tolima have no new injury or suspension concerns to worry about, so they should have a full squad available to them on Wednesday.
Gustavo Ramirez is the one who usually leads their forward line, but he may drop to the bench for Michael Rangel for squad rotation purposes.
America Mineiro, meanwhile, could be without Iago Maidana, who was sent off in their 1-0 defeat over the weekend.
In his place could be Joao Paulo, who will slot into that back-three alongside Eder and German Conti.
At the other end of the pitch, it is likely that Aloisio and Pedrinho will continue to spearhead their attack.
Deportes Tolima possible starting lineup:
Cuesta; Suarez, Mosquera, Caicedo, Angulo; Riascos, Urena; Escobar, Garcia, Orozco; Rangel
America Mineiro possible starting lineup:
Jailson; Paulo, Eder, Conti; Patric, Marlon, Kal, Ramirez, Rodriguinho; Aloisio, Pedrinho
We say: Deportes Tolima 2-0 America Mineiro
With potential qualification as the motivation, the home side should prove to be too strong for Mineiro, who have failed to get off the ground in the Copa Libertadores.