Louis van Gaal's men booked their place in the last eight with a dramatic comeback victory over Mexico in Fortaleza.
Late goals from Wesley Sneijder and Klaas-Jan Huntelaar cancelled out Giovani dos Santos' opener to send the Dutch through, with De Jong's early withdrawal the only low point.
The Milan midfielder hobbled off after just nine minutes, with Van Gaal confirming that the experienced midfielder was all but certain to miss their next game on Saturday.
"De Jong had a small groin strain and I was forced to substitute him," he explained.
"I don't know if he will be available for the quarter-final, though to be honest it's very unlikely he will be able to play the next match.
"He is one of my most important players, so this is an enormous loss for us."
De Jong started each of the Netherlands' group fixtures as they picked up wins against Spain, Australia and Chile.
‘He instantly popped into our ratings as one of the best U21 midfielders in Europe. The impressive thing is Slot has made him into more of a no.6' Inside Ryan Gravenberch's transformation at Liverpool
‘I don’t think Liverpool would look at Ollie Watkins, a striker isn’t a pressing issue for them – it’s Arsenal who need one’ Former Reds star explains why his old club don’t need an out-and-out forward this summer
