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Application of step-wise gradient high-performance counter-current chromatography for rapid preparative separation and purification of diterpene components from Pseudolarix kaempferi Gordon

He, Shichao, Li, Shucai, Yang, Jianhong, Ye, Haoyu, Zhong, Shijie, Song, Hang, Zhang, Yongkui, Peng, Cheng, Peng, Aihua, Chen, Lijuan
Journal of chromatography 2012 v.1235 pp. 34-38
Larix kaempferi, Oriental traditional medicine, alcohols, countercurrent chromatography, polycyclic aromatic hydrocarbons
In general, simultaneously separation and purification of components with a broad polarity range from traditional Chinese medicine (TCM) is a challenge by an ordinary high-speed counter-current chromatography (HSCCC) method. In this paper, we describes a rapid and efficient separation method of combining three-step gradient elution and two-step flow-rate gradient elution using high-performance counter-current chromatography (HPCCC) to separate 8 diterpene compounds simultaneously within 80min in a single run from the alcohol extract of Pseudolarix kaempferi Gordon. This separation process produced 166mg pseudolaric acid B O-β-d-glucopyranoside (PABGly), 152mg pseudolaric acid C (PAC), 8mg deacetylpseudolaric acid A (deacetylPAA), 5mg pseudolaric acid A O-β-d-glucopyranoside (PAAGly), 484mg pseudolaric acid B (PAB), 33mg pseudolaric acid B methyl ester (PAB methyl ester), 10mg pseudolaric acid A (PAA) and 18mg pseudolaric acid H (PAH) from 1.0g crude sample with purities of 98.6%, 99.6%, 92.3%, 92.2%, 99.2%, 99.4%, 98.3%, 91.0%, respectively. Our study indicates that the suitable combination of step-wise gradient elution and flow-rate gradient elution using HPCCC is an effective strategy to separate complex components from natural products.