Hair dyeing, hair perming, and daily hair dryer use can substantially damage hair. Consequently, the demand for products containing natural ingredients for the care of damaged hair is growing. Although polyphenols with antioxidant effects are often used for hair conditioning, few studies have focused on hair conditioning, and the potential of Sparassis latifolia mushroom extract for hair improvement has not been evaluated to date. In this study, the antioxidant activity of and polyphenol content in hot water, 70% and 100% ethanol (EtOH), n-hexane, ethyl acetate (EtOAc), and water extracts of S. latifolia were analyzed. A hair treatment containing S. latifolia extract was prepared, and its effect on damaged hair was evaluated. The highest antioxidant activity was observed in the hot water and EtOAc extracts. Moreover, polyphenol analysis using high-performance liquid chromatography and liquid chromatography-mass spectroscopy confirmed that the EtOAc fraction has relatively high contents of specific polyphenols beneficial for hair. Based on these results, a hair treatment containing S. latifolia extract was applied to damaged hair, and hair improvement was evaluated using hair thickness, tensile strength, and scanning electron microscopy. The hair treatment containing 70% EtOH extract effectively improved hair condition. We postulate that this improvement was caused by the high hydrophobic polyphenol content in the 70% EtOH extract.