The forward had hoped to have the final game of a three-match ban wiped out but the Administrative Court of Sport (CAS) has upheld the original sanctions
The Portuguese was suspended for three games after he was sent off in the 1-1 draw with Athletic Bilbao on February 2 following an altercation with Carlos Gurpegui and Ander Iturraspe.
The 29-year-old has since served two games of that ban having missed the 4-2 win over Villarreal and last weekend's 3-0 victory away to Getafe.
However, Madrid lodged an appeal over the severity of the punishment in the hope Ronaldo would make an earlier than expected return in their clash against Elche.
Blancos coach Carlo Ancelotti included his star man in the squad for Saturday's La Liga fixture at the Santiago Bernabeu, only for the Administrative Court of Sport (CAS) to uphold the ban.
Ronaldo will now return to league action when his side travel to Atletico Madrid for a crunch top-of-the-table clash on March 2.