Mbappe was substituted in the 80th minute of Real Madrid's 3-2 success over Alaves due to an injury problem, but head coach Carlo Ancelotti played down the severity of the issue after the match.
"He's fine. He was a little overloaded and he asked me to change to avoid any problems," Ancelotti told reporters in his post-match press conference.
However, it was reported on Wednesday morning that Mbappe was a major doubt for Sunday's Madrid derby against Atletico Madrid due to the nature of the problem.
Los Blancos have now revealed that the summer arrival damaged the femoral biceps in his left leg against Alaves, and he is now set for a spell on the sidelines.
Real Madrid confirm thigh injury for Mbappe
"Following the tests carried out today on our player Kylian Mbappe by the Real Madrid Medical Services, he has been diagnosed with an injury to the femoral biceps of his left leg. Pending evolution," read a statement from the European and Spanish champions.
Real Madrid have not provided a timeframe when it comes to his recovery, but according to journalist Mario Cortegana, the forward is expected to be out of action for the next three weeks.
The 25-year-old made a slow start to his Real Madrid career, but he has now managed seven goals and one assist in nine appearances for the club in all competitions.
Which games will Mbappe miss for Real Madrid?
If it proves to be a three-week absence for Mbappe, then he would be absent for Real Madrid's next three matches against Atletico Madrid, Lille and Villarreal.
The forward would also not link-up with the France squad during the October international break, with his return to action potentially coming against Celta Vigo in La Liga on October 19.
Mbappe's absence is a huge blow for Real Madrid heading into this weekend's Madrid derby, and it will force Ancelotti into a rethink when it comes to his starting side.
Endrick will be under consideration for his first Real Madrid start, with the 18-year-old the obvious replacement through the middle, but it is more likely that there will be a change of formation.
Jude Bellingham could be used as a number 10 behind Vinicius Junior and Rodrygo - as was the case for the majority of last season - but it is also possible that Arda Guler could be introduced into the XI.
Much will depend on the fitness of Eduardo Camavinga, as the Frenchman is being tipped to return to the squad this weekend, but he has not made a single appearance this term due to the knee injury that he suffered ahead of the UEFA Super Cup in August.