Both sides still have something to play for coming into this one and the clash promises to be an exciting affair.
Match preview
Colon made it back-to-back continental victories by seeing off the threat of Olimpia Asuncion in a 2-1 win on Thursday.
Julio Cesar Falcioni's men fell behind to Hugo Quintana's opener before hitting back thanks to goals in both halves from Federico Lertora.
Consequently, The Sabalero now sit top of Group G with 10 points from five outings but still need all three points on the final day to guarantee their place as group winners.
Standing in their way is an opposing side who will be seeking to make up for a 2-1 defeat in the reverse fixture between the sides last month.
However, Thursday's visitors have failed to win any of their most recent seven away outings, while also failing to score in each of the last two.
Penarol were held to a goalless draw on home turf in their clash against Paraguayan outfit Cerro Porteno on Tuesday.
Mauricio Larriera's men had the best chance to come out on top in what was an open contest as Pablo Ceppelini fluffed his lines from 12 yards just past the hour mark.
The Aurinegros, however, followed that up with a 1-0 triumph over Boston River in their subsequent league outing and now occupy fourth place in the Apertura Division.
Heading into Thursday's match, the home side sit bottom of their Copa Libertadores group but a place in the Copa Sudamericana is still up for grabs should results go their way.
Penarol Copa Libertadores form:
Penarol form (all competitions):
Colon Copa Libertadores form:
Colon form (all competitions):
Team News
Penarol's Juan Ramos is expected to miss Thursday's game after he came off with an injury in the first half of his side's last league outing.
This should see Jairo O'Neil get the nod to feature right from the off as the defender made his return to action in the aforementioned match.
Off the back of a brace in his last outing, Federico Lertora will be looking to come up trumps for Colon once again having recently ended a 17-game goalless streak.
Ramon Abila had to make do with a place on the bench against Asuncion but the 32-year-old is expected to be recalled to the starting XI for this one
Penarol possible starting lineup:
Dawson; O'Neil, Elizalde, Menosse, Aguirregaray; Gargano, Musto; Ceppelini, Viatri, Alonso; Alvarez
Colon possible starting lineup:
Burian; Garces, Goitz, Delgado; Meza, Aliendro, Lertora, Teuten; Bernardi, Farias, Abila