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Separation and purification of quinolyridine alkaloids from seeds of Thermopsis lanceolata R. Br. by conventional and pH-zone-refining counter-current chromatography |
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| Authors: | Fansheng Ning Heng Zhu Huijiao Yan Jiguo Liu Shahid Aziz Hidayat Hussain Xiangyun Song Lei Xie Zhaoqing Meng Guiyun Cao Daijie Wang |
| Affiliation: | 1. Shandong Analysis and Test Center, Qilu University of Technology (Shandong Academy of Sciences), Jinan, P. R. China Biological Engineering Technology Innovation Center of Shandong Province, Heze Branch of Shandong Academy of Sciences, Qilu University of Technology, Heze, P. R. China;2. Shandong Analysis and Test Center, Qilu University of Technology (Shandong Academy of Sciences), Jinan, P. R. China;3. Heze Peony Development Service Center, Heze, P. R. China;4. Shandong Analysis and Test Center, Qilu University of Technology (Shandong Academy of Sciences), Jinan, P. R. China Department of Chemistry, Mirpur University of Science and Technology (MUST), Mirpur, Pakistan;5. Department of Bioorganic Chemistry, Leibniz Institute of Plant Biochemistry, Halle, Germany;6. Shandong Hongjitang Pharmaceutical Group Co., Ltd., Jinan, P. R. China |
| Abstract: | In this work, the preparative separation of quinolyridine alkaloids from seeds of T. lanceolata by conventional and pH-zone-refining counter-current chromatography. Traditional counter-current chromatography separation was performed by a flow-rate changing strategy with a solvent system of ethyl acetate-n-butanol-water (1:9:10, v/v) and 200 mg sample loading. Meanwhile, the pH-zone-refining mode was adopted for separating 2.0 g crude alkaloid extracts with the chloroform-methanol-water (4:3:3, v/v) solvent system using the stationary and mobile phases of 40 mM hydrochloric acid and 10 mM triethylamine. Finally, six compounds, including N-formylcytisine (two conformers) ( 1 ), N-acetycytisine (two conformers) ( 2 ), (-)-cytisine ( 3 ), 13-β-hydroxylthermopsine ( 4 ), N-methylcytisine ( 5 ), and thermopsine ( 6 ) were successfully obtained in the two counter-current chromatography modes with the purities over 96.5%. Moreover, we adopted nuclear magnetic resonance and mass spectrometry for structural characterization. Based on the obtained findings, the pH-zone-refining mode was the efficient method to separate quinolyridine alkaloids relative to the traditional mode. |
| Keywords: | alkaloids conventional and pH-zone-refining mode counter-current chromatography separation method Thermopsis lanceolata R. Br |