鹿茸草中3种苯乙醇苷的分离制备工艺研究
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摘要
目的利用大孔吸附树脂以及动态轴向分离技术,建立鹿茸草Monochasma savatieri中3种苯乙醇苷类化合物麦角甾苷(acteoside)、异麦角甾苷(isoacteoside)、torenoside B的最佳分离制备工艺。方法通过静态吸附与解吸附实验和动态分离实验,研究8种常用大孔树脂AB-8、D101、HPD100、LSA-10、LX-11、LX-17、LX-38、XDA-6对该类化合物的分离特性,选出最佳大孔树脂类型;采用所选最佳大孔树脂对3种苯乙醇苷类化合物的纯化工艺参数进行优化。经放大实验得到总苯乙醇苷,再经动态轴向分离得到3种苯乙醇苷类化合物单体。结果优化出的最理想大孔树脂为LX-17;优选的最佳上样质量浓度为原药材1.8 g/m L;最佳吸附时间为150 min;洗脱条件为乙醇-水系统,体积流量为2.5 m L/min,梯度浓度依次为纯水(4 BV)、15%乙醇(4 BV)、60%乙醇(5 BV)以及90%乙醇(2 BV);随后放大10倍进行中试,得到总苯乙醇苷部位,得率为5.25%,其中麦角甾苷15.24%、异麦角甾苷32.14%、torenoside B 29.33%;5 g总苯乙醇苷部位经动态轴向压缩制备柱纯化出3种苯乙醇苷单体成分,分别为麦角甾苷0.127 g,得率2.54%,质量分数为92%;异麦角甾苷0.894 g,得率17.88%,质量分数为95%;torenoside B 0.962 g,得率19.24%,质量分数为98%。结论 LX-17型大孔树脂可作为纯化苯乙醇苷的较佳分离材料,经过优化的分离工艺稳定可靠,有望在工业化生产中推广应用。
Objective The optimal simultaneous separation process of three phenylethanoid glycosides(acteoside, isoacteoside, and torenoside B) from Monochasma savatieri were established using of macrophage resins and dynamic axial column(DAC). Methods The adsorption/desorption experiments and dynamic separation experiments were performed on eight types of resins(AB-8, D 101, HPD 100, LSA-10, LX-11, LX-17, LX-38, and XDA-6) to find the optimal resin. Then the optimal separation parameters were investigated on the chosen resin. The total phenylethanoid glycosides obtained from the large-scale experiment were further seperated to get acteoside, isoacteoside, and torenoside B, respectively using of DAC system. Results Among these candidate resins, LX-17 was chosen to further obtain the optimal parameters: The optimal feeding concentration of raw materials was 1.8 g/m L; The optimal adsorption time was 150 min; The optimal gradient elute conditions were ethanol/water(0/100, 4 BV; 15/85, 4 BV; 60/40, 5 BV; 90/10, 2 BV). The large-scale experiments were amplified to 10 folds on the basis of optimal parameters to obtain total phenylethanoid glycosides. Acteoside, isoacteoside, and torenoside B were simultaneously obtained from total phenylethanoid glycosides using DAC system. Conclusion LX-17 and DAC system can be used for the purification of phenylethanoid glycosides, which will have a good future for the application in industry.
Objective The optimal simultaneous separation process of three phenylethanoid glycosides(acteoside, isoacteoside, and torenoside B) from Monochasma savatieri were established using of macrophage resins and dynamic axial column(DAC). Methods The adsorption/desorption experiments and dynamic separation experiments were performed on eight types of resins(AB-8, D 101, HPD 100, LSA-10, LX-11, LX-17, LX-38, and XDA-6) to find the optimal resin. Then the optimal separation parameters were investigated on the chosen resin. The total phenylethanoid glycosides obtained from the large-scale experiment were further seperated to get acteoside, isoacteoside, and torenoside B, respectively using of DAC system. Results Among these candidate resins, LX-17 was chosen to further obtain the optimal parameters: The optimal feeding concentration of raw materials was 1.8 g/m L; The optimal adsorption time was 150 min; The optimal gradient elute conditions were ethanol/water(0/100, 4 BV; 15/85, 4 BV; 60/40, 5 BV; 90/10, 2 BV). The large-scale experiments were amplified to 10 folds on the basis of optimal parameters to obtain total phenylethanoid glycosides. Acteoside, isoacteoside, and torenoside B were simultaneously obtained from total phenylethanoid glycosides using DAC system. Conclusion LX-17 and DAC system can be used for the purification of phenylethanoid glycosides, which will have a good future for the application in industry.
引文
[1]潘丽,申琳.肠炎宁颗粒治疗小儿消化不良的临床观察[J].中草药,2016,47(12):2147-2151.
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[16]Sun Y,Yuan H Y,Hao L L,et al.Enrichment and antioxidant properties of flavone C-glycosides from trollflowers using macroporous resin[J].Food Chem,2013,141(1):533-541.
[17]Dong Y,Zhao M M,Sun-Waterhouse D X,et al.Absorption and desorption behaviour of the flavonoids from Glycyrrhiza glabra L.leaf on macroporous adsorption resins[J].Food Chem,2015,168:538-545.
[18]Liu B Y,Ouyang J,Yuan X F,et al.Adsorption properties and preparative separation of phenylethanoid glycosides from Cistanche deserticola by use of macroporous resins[J].J Chromatogr B,2013,937:84-90.
[19]Sun Y,Yuan H Y,Hao L L,et al.Enrichment and antioxidant properties of flavone C-glycosides from trollflowers using macroporous resin[J].Food Chem,2013,141(1):533-541.
[20]Wang L,Xu Q,Su S,et al.Simultaneous purification of pulchinenoside B-4 and B-5 from Pulsatilla chinensis using macroporous resin and preparative high performance liquid chromatography[J].Ind Eng Chem Res,2012,51(45):14859-14866.
[21]Sakuma S,Motomura H.Purification of saikosaponin-a,saikosaponin-C and saikosaponin-D-application of large-scale reversed-phase high-performance liquidchromatography[J].J Chromatogr A,1987,400:293-295.
[22]Medina I.Purification of zopiclone by preparative high performance liquid chromatography[J].J Liq Chromatogr R T,1998,21(17):2689-2698.
[2]肖培根.新编中药志(一)[M].北京:化学工业出版社,2001.
[3]国家中医药管理局《中华本草》编委会.中华本草(上册)[M].上海:上海科技出版社,1998.
[4]Zheng W,Tan X Q,Guo L J,et al.Chemical constituents from Monochasma savatieri[J].Chin J Nat Med,2012,10(2):102-104.
[5]师梦凡.鹿茸草中苯乙醇苷类化学成分的心肌保护作用研究[D].苏州:苏州大学,2013.
[6]朱莹莹.鹿茸草中苯乙醇苷的分离制备及抗菌活性研究[D].苏州:苏州大学,2013.
[7]Li M,Shi M F,Liu Y L,et al.Phenylethanoid glycosides from Monochasma savatieri and their anticomplement activity through the classical pathway[J].Planta Med,2012,78(12):1381-1386.
[8]Jiménez C,Riguera R.Phenylethanoid glycosides in plants:Structure and biological activity[J].Nat Prod Rep,1995,11(6):591-606.
[9]Pan J,Yuan C S,Lin C J,et al.Pharmacological activities and mechanisms of natural phenylpropanoid glycosides[J].Pharmazie,2003,58(11):767-775.
[10]Noel J P,Austin M B,Bomat E K.Structure-function relationships in plant phenylpropanoid biosynthesis[J].Curr Opin Plant Biol,2005,8(3):249-253.
[11]Li C Q,Li B G,Qi H Y,et al.Three cyclooctapeptides and one glycoside from Microtoena prainiana[J].J Nat Prod,2004,67(6):978-982.
[12]曹馨元,冯霞,李茂星,等.甘肃马先蒿毛蕊花糖苷和异毛蕊花糖苷制备工艺研究[J].中草药,2016,47(10):1696-1701.
[13]崔翰明,张秋燕,林海,等.三七总皂苷的大孔吸附树脂纯化工艺和质量分析研究[J].中草药,2012,43(11):2177-2182.
[14]贾存勤,李阳春,屠鹏飞,等.HPD系列大孔吸附树脂预处理方法研究[J].中国中药杂志,2005,30(18):1425-1427.
[15]孙宗喜,徐桂花,李艳芳,等.大孔树脂纯化绵茵陈中总有机酸和绿原酸的研究[J].中药材,2012,35(2):306-310.
[16]Sun Y,Yuan H Y,Hao L L,et al.Enrichment and antioxidant properties of flavone C-glycosides from trollflowers using macroporous resin[J].Food Chem,2013,141(1):533-541.
[17]Dong Y,Zhao M M,Sun-Waterhouse D X,et al.Absorption and desorption behaviour of the flavonoids from Glycyrrhiza glabra L.leaf on macroporous adsorption resins[J].Food Chem,2015,168:538-545.
[18]Liu B Y,Ouyang J,Yuan X F,et al.Adsorption properties and preparative separation of phenylethanoid glycosides from Cistanche deserticola by use of macroporous resins[J].J Chromatogr B,2013,937:84-90.
[19]Sun Y,Yuan H Y,Hao L L,et al.Enrichment and antioxidant properties of flavone C-glycosides from trollflowers using macroporous resin[J].Food Chem,2013,141(1):533-541.
[20]Wang L,Xu Q,Su S,et al.Simultaneous purification of pulchinenoside B-4 and B-5 from Pulsatilla chinensis using macroporous resin and preparative high performance liquid chromatography[J].Ind Eng Chem Res,2012,51(45):14859-14866.
[21]Sakuma S,Motomura H.Purification of saikosaponin-a,saikosaponin-C and saikosaponin-D-application of large-scale reversed-phase high-performance liquidchromatography[J].J Chromatogr A,1987,400:293-295.
[22]Medina I.Purification of zopiclone by preparative high performance liquid chromatography[J].J Liq Chromatogr R T,1998,21(17):2689-2698.