Hypericin has shown promising potential for the prevention and treatment of cancer. Here we report facile strategy for the separation and purification of natural products from the extracts by demonstrating hypericin isolation from Hypericum perforatum L. extract using macroporous resin column chromatography (MRCC) fol-lowed by preparative liquid chromatography (PLC). Effect of various parameters including the kinds of resins, loading extract concentration, loading flow rate, types of eluent, and elution flow rate on the separation effi-ciency of MRCC for hypericin was investigated. The purity and purification ratio of hypericin was 71.8 % and 2.4 respectively, optimized by XAD16 N MRCC with a high stability and reproducibility. PLC parameters were determined by optimizing stationary phase, mobile phase, and injection sample amount. The high purity hypericin (98.7 %) was achieved by PLC and the total recovery ratio of hypericin was 80.6 % in the coupling process.