The 37-year-old singer performed at the City of Hope gala just days after wrapping her "On the Run II" tour with husband Jay-Z.
Dressed in a shimmering Ralph & Russo feathered gown, the mother of three performed a three-song set at the event, which also honoured outgoing Warner/Chappell Music Chairman Jon Platt, both the singer and Jay's longtime publisher.
According to editors at USA Today, she performed songs "XO", "Perfect" and "Halo" to a crowd of 1,200 at the benefit and was introduced by Platt.
"I was reminded even when you're the biggest talent in world, it still comes down to hard work," the music executive said, referring to seeing her on tour.
"She leads by example. She shows all of us how to dream big," added Platt, who is expected to take the helm at Sony/ATV Music Publishing in April.
And Jay returned the favour by introducing the night's Spirit of Life honouree as his "brother" and "the Obama of the music industry."
Pharrell hosted the event, which benefited City of Hope, an independent research and treatment centre for cancer, diabetes and other life-threatening diseases. In its 45-year history, the organisation has raised more than $118 million (£89 million) and honoured key figures in the music and entertainment industry, including Quincy Jones and Clive Davis.
"Beyonce agreed to perform without a moment's hesitation, and that wasn't just for me," Platt told Variety last month. "She wanted to be part of our celebration for the patients, doctors, scientists, and nurses who make City of Hope a place where miracles can happen."
"That's what makes her a cultural icon. She's not only a genius, in a class of her own, but she has phenomenal heart and soul, and she knows how to use them."