Abstract: OBJECTIVE To prepare highly purified proanthocyanidins from Ginkgo biloba L., so as to provide material base for the systematic study of proanthocyanidins from Ginkgo biloba L.. METHODS The purification processes of macroporous resins and polyamide resin were optimized by using NP-HPLC methods; qualitative and quantitative analysis of flavonoid, the impurity of Ginkgo biloba proanthocyanidin extract(GPE), was realized by HPLC with different chromatographic conditions; the anti-oxidant activities of proanthocyanidins from Ginkgo biloba L. and grape seed were compared by DPPH assay. RESULTS The purity of proanthocyanidins in GPE was >90%, and the content of flavonoid was only 3.44%. The GPE exhibited higher DPPH radical scavenging activity with IC₅₀ 22.87 μg·mL^-1 than that of grape-seed proanthocyanidins with IC₅₀ 36.99 μg·mL^-1. CONCLUSION The process of this study could be prepared to obtain high-purity GPE, and its antioxidant activity is better than grape-seed proanthocyanidin.