Madrid manager Zinedine Zidane had included Hazard in his squad to face Real Valladolid after saying the Belgian was finally "ready" to make his first appearance of the new season.
Hazard is expected to miss another three-to-four weeks, according to Spanish daily Marca.
The 29-year-old last played in an Aug. 7 loss to Manchester City in the Champions League.
He's made just 22 appearances in all competitions since joining Los Blancos in July 2019 for a reported €100 million.
Hamstring, ankle, leg, and foot injuries have plagued the Belgian winger since his arrival from Chelsea.
Zidane said Tuesday he was happy with his team, but Hazard's injury could force Madrid to seek relief over the remaining days of the summer transfer window.
Martin Odegaard, however, was recalled from loan.