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The aim of this study is to assess how different extraction temperatures affect the in vitro antioxidant potential of polysaccharides derived from the leaves of Myxopyrum smilacifolium. The findings indicated that, the polysaccharides exhibit the highest antioxidant activity when extracted at a temperature of 90°C. Fourier transform infrared spectroscopy analysis confirmed the presence of characteristic absorption groups typically found in polysaccharide structures in the PS-T90 sample. The average molecular weight of the extracted polysaccharides was approximately 9.30×105 Da. The total antioxidant capacity of PS-T90 was determined to be 0.2646 ± 0.0007 mg GA/g or 0.1725 ± 0.0007 mg AS/g. Additionally, the IC50 values for the polysaccharides against 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS) radicals were measured to be 1.04 mg/mL and 3.37 mg/mL, respectively. These exceptional bioactive properties suggest the potential of PS-T90 as a valuable resource for antioxidant development.