The video was also shared on Twitter, as concerned fans speculated what might have happened.
While many fans voiced concerns online, others speculated that the eye twitching could be due to false eyelash glue sticking her eye shut.
WATCH: Katy Perry's unborn baby gives her the finger
Other reasons that fans speculated were that her eyes could be dry, that she may have been suffering from a migraine, or that she could have had an eye spasm due to stress or fatigue.
Last year on American Idol, Katy Pery revealed that she suffers from ‘wonks eye’ which some fans think may be the culprit.
"I have a wonk eye as well, and I used to be worried about it," she shared on American Idol.
"Then a bunch of my fans created a fandom over my wonk-eye. I even have a fandom that calls itself, 'Katy's wonk-eye.'"
The pop star continued the performance without addressing her eye and is yet to respond to concerned fans.