The team with the most points at the end of the season crowned champions.
The teams had to declare their interest to participate in the 2001 UEFA Intertoto Cup before the end of the championship.
At the end of the championship, the higher placed team among the interested ones participated in the Intertoto Cup (if they had not secured their participation in any other UEFA competition).
Ethnikos Assia, Anagennisi Deryneia and Alki Larnaca were relegated from previous season and played in the 2000–01 Cypriot Second Division.
They were replaced by the first three teams of the 1999–2000 Cypriot Second Division, Digenis Morphou, Aris Limassol and Doxa Katokopias.