Abstract: Objective To study the preparing technology of 3,5-O-dicaffeoyl quinic acid with high purity and its antiproliferative activity on HeLa cells. Methods 3,5-O-dicaffeoyl quinic acid was prepared by column chromatographic extraction methods coupled with medium pressure liquid chromatography from flower of Artemisia anomala. The antiproliferative activity of 3,5-O-dicaffeoyl quinic acid on HeLa cell was evaluated by MTT method. Results The extraction and purification recovery yield were 99.0% and 61.2%, respectively. The total recovery yield of 3,5-O-dicaffeoyl quinic acid reached 54.0%. With the increase of 3,5-O-dicaffeoyl quinic acid concentration, the HeLa cells viability decreased and the cell morphological damage increased. The IC₅₀ value of 3,5-O-dicaffeoyl quinic acid on HeLa cells was 26.5 µg·mL^-1. Conclusion A simple, efficient, energy-saving 3,5-O-dicaffeoyl quinic acid preparation method is developed. The compound displays good antiproliferative activity on HeLa cells in vitro.