仙茅多糖的分离纯化及结构分析
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摘要
目的:对仙茅多糖进行分离纯化,并对其结构进行分析,为仙茅多糖的进一步应用提供理论基础。方法:仙茅根茎经水提、醇沉、冷冻干燥得仙茅粗多糖(XMP),经DEAE-纤维素柱和Sepharose CL-6B凝胶柱色谱纯化得纯多糖XMP-1,采用凝胶渗透色谱、紫外光谱、红外光谱、气质联用等技术对XMP-1的结构进行分析。结果:仙茅多糖由甘露糖、葡萄糖、半乳糖3种单糖组成,其摩尔比为81.2∶5.7∶1,多糖质量分数为91.8%,相对分子质量为3 031 Da,不含核酸、蛋白质和色素。具有吡喃特征吸收峰,在水溶液分子中的分子构象可能为自然卷曲结构。结论:仙茅纯多糖是一种分子量较小的中性杂多糖。
Objective: To study the isolation,puriffication and structural analysis of the polysaccharides from the Curculiginis Rhizoma. The work will provide the theoretical foundation for the further application of Curculiginis Rhizoma polysaccharides. Method: The crude polysaccharide was obtained by hot water extraction,ethanol precipitation and freezedrying. After further purification by DEAE-cellulose and Sepharose CL-6B column chromatography,pure polysaccharide( XMP-1) was achieved. The structural of XMP-1 was elucidated by GPC,UV,IR and GC-MS. Result: XMP-1 was composed of mannose,glucose,galactose with a molar ratio of 81. 2∶5. 7∶1. Total carbohydrate content of XMP-1 was 91. 8% and the molecular weight was 3 031 Da. XMP-1 have no nucleic aicd,protein and pigment. IR chromatography showed that XMP-1 has pyrane characteristic absorption peak. XMP-1 in molecular conformation in aqueous solution can be naturally curly structure. Conclusion: XMP-1is a kind of small molecular weight neutral pure heterosaccharides.
Objective: To study the isolation,puriffication and structural analysis of the polysaccharides from the Curculiginis Rhizoma. The work will provide the theoretical foundation for the further application of Curculiginis Rhizoma polysaccharides. Method: The crude polysaccharide was obtained by hot water extraction,ethanol precipitation and freezedrying. After further purification by DEAE-cellulose and Sepharose CL-6B column chromatography,pure polysaccharide( XMP-1) was achieved. The structural of XMP-1 was elucidated by GPC,UV,IR and GC-MS. Result: XMP-1 was composed of mannose,glucose,galactose with a molar ratio of 81. 2∶5. 7∶1. Total carbohydrate content of XMP-1 was 91. 8% and the molecular weight was 3 031 Da. XMP-1 have no nucleic aicd,protein and pigment. IR chromatography showed that XMP-1 has pyrane characteristic absorption peak. XMP-1 in molecular conformation in aqueous solution can be naturally curly structure. Conclusion: XMP-1is a kind of small molecular weight neutral pure heterosaccharides.
引文
[1]国家药典委员会.中华人民共和国药典.一部[S].北京:中国医药科技出版社,2010:94.
[2]黄有霖.仙茅的研究进展[J].中药材,2003,26(3):225-228.
[3]Wang Z H,Huang J,Ma X C,et al.Phenolic glycosides from Curculigo orchioides[J].Fitoterapia,2013,86(7):64-69.
[4]Zuo A X,Shen Y,Jiang Z Y,et al.Two new triterpenoid glycosides from Curculigo orchioides[J].Asian Nat Prod Res,2012,14(5):407-412.
[5]Zuo A X,Shen Y,Jiang Z Y,et al.Three new phenolic glycosides from Curculigo orchioides[J].Fitoterapia,2010,81(7):910-913.
[6]Zuo A X,Shen Y,Zhang X M,et al.Four new trace phenolic glycosides from Curculigo orchioides[J].Asian Nat Prod Res,2010,12(1):43-50.
[7]Dall'Acqua S,Shrestha B B,Comai S,et al.Two phenolic glycosides from Curculigo orchioides[J].Fitoterapia,2009,80(5):279-282.
[8]季春.仙茅多糖COPb-1和COPf-1的分离纯化及结构研究[J].贵州化工,2005,30(1):17-19.
[9]Brown G D,Gordon S.Immune recognition:a new receptor for beta-glucans[J].Nature,2001,413(6851):36-37.
[10]Brown G D,Herre J,Williams D L,et al.Dectin-1mediates the biological effects of betaglucans[J].Exp Med,2003,197(9):1119-1124.
[11]Gantner B N,Simmons R M,Canavera S J,et al.Collaborative induction of inflammatory responses by dectin-1 an Toll-like receptor 2[J].Exp Med,2003,197(9):1107.
[12]韩艳萍,赵鲁杭,吴海明.壳寡糖激活巨噬细胞的机制[J].浙江大学学报:医学版,2006,35(3):265-268.
[13]邵力成,尹登科,高向东.天然多糖免疫细胞受体的研究进展[J].药物生物技术,2006,13(5):389-392.
[14]彭梅,唐建波,肖雄,等.提取方法对仙茅多糖提取率及抗肿瘤活性的影响[J].中成药,2014,36(9):1985-1988.
[15]石磊.几种多糖的分离纯化、结构研究和生物活性研究[D].济南:山东大学,2007.
[16]颜军,易勇,邬晓勇,等.黄芪多糖的相对分子量测定及单糖组成分析[J].食品科技,2012,37(12):278-283.
[17]孙晓燕,蔡昌利,徐丽莉,等.多糖含量测定方法的比较[J].现代中药研究与实践,2015,29(3):58-62.
[18]陈巧巧,万琴,王振中,等.人参多糖中糖醛酸含量测定方法的建立[J].中国实验方剂学杂志,2012,18(8):121-124.
[19]陈帅,许程剑,李应彪,等.阿魏菇多糖的结构分析[J].现代食品科技,2015,31(3):29-37.
[20]马小双,李程程.不同种类铁皮石斛及其多糖的红外光谱测定分析[J].黑龙江农业科学,2015(9):116-118.
[21]石磊,韩龙,刘超.多糖的构象研究方法综述[J].曲阜师范大学学报:自然科学版,2012,38(3):78-84.
[22]徐航,朱锐,刘玮,等.多糖高级结构解析方法的研究进展[J].药学进展,2015,39(5):364-369.
[23]邹胜,徐溢,张庆.天然植物多糖分离技术研究现状和进展[J].天然产物研究与开发,2015,27(8):1051-1059.
[24]黄菊清.竹茹多糖的化学结构和免疫活性研究[D].杭州:浙江大学,2015.
[25]陈海霞.活性多糖的结构与效应关系[J].科学观察,2013,8(6):53-55.
[26]周勇,张丽,赵离原,等.仙茅多糖对小鼠免疫功能调节作用实验研究[J].上海免疫学杂志,1996,16(6):336-338.
[27]余晓红.仙茅多糖对小鼠免疫功能影响的实验研究[J].海峡药学,2011,23(3):33-35.
[28]Zhao T,Feng Y,Li J,et al.Schisandra polysaccharide evokes immunomodulatory activity through TLR 4-mediated activation of macrophages[J].Int J Biol Macromol,2014,65(5):33-40.
[29]芦静波,陈靠山,曹剑峰,等.多糖抗肿瘤活性及其机制研究[J].医学理论与实践,2015,28(22):3048-3050.
[30]刘洁,李文香,王文亮,等.多糖空间结构与生物活性相关性研究进展[J].农业机械,2011(17):153-155.
[2]黄有霖.仙茅的研究进展[J].中药材,2003,26(3):225-228.
[3]Wang Z H,Huang J,Ma X C,et al.Phenolic glycosides from Curculigo orchioides[J].Fitoterapia,2013,86(7):64-69.
[4]Zuo A X,Shen Y,Jiang Z Y,et al.Two new triterpenoid glycosides from Curculigo orchioides[J].Asian Nat Prod Res,2012,14(5):407-412.
[5]Zuo A X,Shen Y,Jiang Z Y,et al.Three new phenolic glycosides from Curculigo orchioides[J].Fitoterapia,2010,81(7):910-913.
[6]Zuo A X,Shen Y,Zhang X M,et al.Four new trace phenolic glycosides from Curculigo orchioides[J].Asian Nat Prod Res,2010,12(1):43-50.
[7]Dall'Acqua S,Shrestha B B,Comai S,et al.Two phenolic glycosides from Curculigo orchioides[J].Fitoterapia,2009,80(5):279-282.
[8]季春.仙茅多糖COPb-1和COPf-1的分离纯化及结构研究[J].贵州化工,2005,30(1):17-19.
[9]Brown G D,Gordon S.Immune recognition:a new receptor for beta-glucans[J].Nature,2001,413(6851):36-37.
[10]Brown G D,Herre J,Williams D L,et al.Dectin-1mediates the biological effects of betaglucans[J].Exp Med,2003,197(9):1119-1124.
[11]Gantner B N,Simmons R M,Canavera S J,et al.Collaborative induction of inflammatory responses by dectin-1 an Toll-like receptor 2[J].Exp Med,2003,197(9):1107.
[12]韩艳萍,赵鲁杭,吴海明.壳寡糖激活巨噬细胞的机制[J].浙江大学学报:医学版,2006,35(3):265-268.
[13]邵力成,尹登科,高向东.天然多糖免疫细胞受体的研究进展[J].药物生物技术,2006,13(5):389-392.
[14]彭梅,唐建波,肖雄,等.提取方法对仙茅多糖提取率及抗肿瘤活性的影响[J].中成药,2014,36(9):1985-1988.
[15]石磊.几种多糖的分离纯化、结构研究和生物活性研究[D].济南:山东大学,2007.
[16]颜军,易勇,邬晓勇,等.黄芪多糖的相对分子量测定及单糖组成分析[J].食品科技,2012,37(12):278-283.
[17]孙晓燕,蔡昌利,徐丽莉,等.多糖含量测定方法的比较[J].现代中药研究与实践,2015,29(3):58-62.
[18]陈巧巧,万琴,王振中,等.人参多糖中糖醛酸含量测定方法的建立[J].中国实验方剂学杂志,2012,18(8):121-124.
[19]陈帅,许程剑,李应彪,等.阿魏菇多糖的结构分析[J].现代食品科技,2015,31(3):29-37.
[20]马小双,李程程.不同种类铁皮石斛及其多糖的红外光谱测定分析[J].黑龙江农业科学,2015(9):116-118.
[21]石磊,韩龙,刘超.多糖的构象研究方法综述[J].曲阜师范大学学报:自然科学版,2012,38(3):78-84.
[22]徐航,朱锐,刘玮,等.多糖高级结构解析方法的研究进展[J].药学进展,2015,39(5):364-369.
[23]邹胜,徐溢,张庆.天然植物多糖分离技术研究现状和进展[J].天然产物研究与开发,2015,27(8):1051-1059.
[24]黄菊清.竹茹多糖的化学结构和免疫活性研究[D].杭州:浙江大学,2015.
[25]陈海霞.活性多糖的结构与效应关系[J].科学观察,2013,8(6):53-55.
[26]周勇,张丽,赵离原,等.仙茅多糖对小鼠免疫功能调节作用实验研究[J].上海免疫学杂志,1996,16(6):336-338.
[27]余晓红.仙茅多糖对小鼠免疫功能影响的实验研究[J].海峡药学,2011,23(3):33-35.
[28]Zhao T,Feng Y,Li J,et al.Schisandra polysaccharide evokes immunomodulatory activity through TLR 4-mediated activation of macrophages[J].Int J Biol Macromol,2014,65(5):33-40.
[29]芦静波,陈靠山,曹剑峰,等.多糖抗肿瘤活性及其机制研究[J].医学理论与实践,2015,28(22):3048-3050.
[30]刘洁,李文香,王文亮,等.多糖空间结构与生物活性相关性研究进展[J].农业机械,2011(17):153-155.