Huntelaar was hurt during a morning training session and immediately left the field to ice his leg. He did not return to training, the Royal Dutch Soccer Association said on its Web site. The extent of the injury was not immediately known.
First-choice striker Ruud van Nistelrooy and backup forward Danny Koevermans trained separately from the team because of slight injuries, but are expected to be available for Saturday's match. Coach Marco van Basten is also missing the injured Arjen Robben and Robin van Persie.
Four other players Wesley Sneijder of Real Madrid, Demy de Zeeuw of AZ Alkmaar, Wilfred Bouma of Aston Villa and Kew Jaliens of AZ have recovered from minor knocks and went through full training. They are expected to be fit for Saturday's match in Rotterdam and the team's final qualifier against Belarus in Minsk next Wednesday.
Van Basten's team is second in Group G behind Romania and needs just two points from its last two matches to qualify for next year's tournament.