He will not travel to South Africa with the rest of the party but has not been ruled out for the World Cup finals which start on Friday (June 11).
"He made an unnatural move and then sustained the hamstring injury," coach Bert van Marwijk told news conference.
"He will not join us tonight on our way to South Africa as he will undergo a scan tomorrow as it was no use to do it now."
Van Marwijk said he had feared injuries from the Netherlands' final warmup match but had started to relax until second-half substitute Robben hobbled off in obvious pain two minutes from time.
"My first thought was this can't be true, the end the final warm-up match, everyone is fit and then this," he added.
"We now have to wait. I can replace Robben until 24 hours before our opening match but as long as there is hope he recovers in time he will stay in the squad."
Until Robben's injury scare, prospects ahead of the June 11-July 11 finals only looked bright for the Dutch who had surprisingly conceded a sixth minute lead to the Hungarians through Balazs Dzsudzsak.
