2000–01 Croatian First Football League

Dinamo Zagreb were the defending champions, having won their fifth consecutive title the previous season.

Marsonia had returned to top flight after one previous three-season spell in the Prva HNL between 1994 and 1997, while Čakovec saw its top flight debut after coming close to promotion in 1998 and 1999 (they lost the promotion play-offs on both occasions).

Those should have been top four clubs, but since third-placed Croatia Sesvete and sixth-placed Belišće had decided to step back from promotion, 1st, 2nd, 4th and 5th placed second level teams were automatically promoted for the following season (Kamen Ingrad, Pomorac Kostrena, Zadar and TŠK Topolovac respectively).

Therefore, the 12th placed Marsonia played a two-legged relegation play-off against the 7th placed team of Druga HNL, Solin.

The tie ended in a 5–5 aggregate score and Marsonia won it on away goals, thereby staying in the Prva HNL for the following season.