Gary Cahill was forced off in the first half of Chelsea's Premier League clash with Leicester City on Saturday due to a hamstring injury.
Club captain Cahill returned to Antonio Conte's starting XI after sitting out the EFL Cup semi-final first-leg draw against Arsenal on Wednesday, but lasted just 33 minutes.
The England international pulled up after racing Jamie Vardy for a high ball over the Chelsea defence and hobbled off the pitch holding his right leg.
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Conte turned to Andreas Christensen as his replacement, forcing David Luiz to wait even longer for his first Premier League appearance since November.
Cahill's injury will be a concern for the Blues with an FA Cup replay against Norwich City and the second leg of the EFL Cup semi-final against Arsenal to come this month, while a Champions League last-16 tie against Barcelona is on the horizon.