Objective: To prepare high purity of hypericin by polymer nanospheres. Method: Hypericin extract was enriched by PEK nanospheres

then PDCX nanospheres further purification

high-purity of hypericin was prepared by ODS column

single factor tests were adopted to optimize purification technology.HPLC was employed to determined the content of hypericin with mobile phase of methanol-0.006 mol·L-1 Na2HPO4(7:1

added H3PO4 adjusted pH to 6.5)and detection wavelength of 590 nm. Result: Static saturated adsorption capacity of PEK nanospheres for hypericin was 8.5 mg·g-1 with elution ratio of 89.6%

after extract was enriched by PEK nanospheres column

purity of hypericin was 3.2% with recovery rate of 89.9%;Purity of hypericin was 33.0% through PDCX nanospheres chromatography

recovery rate was 80.1%;Hypericin with a purity of 91.6% was obtained by ODS column chromatography

recovery rate was 86.7%. Conclusion: This preparation process was efficient and economical

which was suitable for separation and purification of hypericin and other unstable ingredients.