Avianca will begin its planned service from Colombian capital Bogota to Santa Cruz in Bolivia on 31 October.
The Star Alliance airline will operate daily on the route, with Airbus A320 aircraft configured for 150 passengers including 12 in business class.
Avianca chief executive Hernan Rincon told FlightGlobal in August that the airline was awaiting Bolivian regulatory approval for the route.
The carrier already serves Santa Cruz from Peru's capital Lima, and also operates to another Bolivian city, La Paz, from both Lima and Bogota.
No other airline operates nonstop between Bogota and Santa Cruz, FlightGlobal schedules data shows.