The 2000–01 Argentine Primera B Nacional was the 15th season of second division professional of football in Argentina.
A total of 30 teams competed; the champion and runner-up were promoted to Argentine Primera División.
The winning team was declared champion and was automatically promoted to Primera División.
Note: Nueva Chicago won 4–2 on aggregate, promoting to Primera División.
The best two teams of each zone that were not promoted (Quilmes and Instituto) played against the 18th and the 17th placed of the Relegation Table of 2000–01 Primera División.