The 63-year-old television personality denied in an interview with The Hollywood Reporter's "Awards Chatter" podcast that she will oppose President Donald Trump in the 2020 election if the businessman seeks a second term.
"I will never run for public office. That's a pretty definitive thing," Winfrey said after host Scott Feinberg asked if she'd be willing to take on Trump.
"I don't know [if I would beat him]," she added. "I don't know the answer to that. But I will never have to know the answer to that, because I will never run for public office."
Winfrey supported former Secretary of State Hillary Clinton in the 2016 election.
She said on "The David Rubenstein Show" in March that she reconsidered her chance at becoming president in the wake of Trump's win.
"I actually never thought that was [an option]. I never considered the question even a possibility," the star explained.
"I thought, 'Oh, gee, I don't have the experience. I don't know enough.' Now I'm thinking, 'Oh,'" she added with a smile.
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