De Jong will miss the Champions League last 16 against Paris Saint-Germain or Newcastle as well as La Liga fixtures with Villarreal, Athletic Bilbao, Sevilla and Rayo Vallecano.
The 28-year-old will also be absent for next Tuesday's Spanish Cup semi-final second leg as the Catalans look to overturn a 4-0 deficit against Atletico Madrid.
In a short statement on their website, Barcelona said De Jong had injured his "right hamstring during training this morning and will be out for five to six weeks".
Barca are just a point clear of Real Madrid atop the La Liga table before hosting Villarreal on Saturday.
De Jong is set to be sidelined for the March international window as the Oranje face Norway and Ecuador in pre-World Cup friendlies.