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Heteroclitin D (H.D) was successfully isolated from Kadsurae Caulis by using flash chromatography and recrystallized by methanol, 10.2mg of H.D was obtained from 4.86g of crude extract, and the purity determined by HPLC was 99.4%. The structure was identified by UV, IR, MS, and NMR analysis. The fast, simple and efficient method can be applied to the preparation of reference substance of H. D.