三萜及其皂苷的结构研究_沈药研究生课.ppt

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,三萜及其皂苷的结构研究,沈阳药科大学天然药化,宋少江,一、概述,二、结构解析规律,三、结构解析实例,四、小结,一、概述,三萜皂苷由三萜皂苷元与糖或糖醛酸组成。,糖的组成主要有葡萄糖、半乳糖、鼠李糖、阿拉伯糖、以及其它戊糖类。,常见的糖醛酸主要有葡萄糖醛酸及半乳糖醛酸等。,苷元主要包括四环三萜（如人参皂苷）及五环三萜（如甘草皂苷）。,分配柱色谱,法要比吸附柱色谱法好，常用硅胶为支持剂。,(,薄层硅胶多加压,),常用溶剂系统,：,氯仿：甲醇：水（,6,：,4,：,1;7,：,3,：,0.5;,8,：,2,：,0.25,等单一系统或梯度洗脱）,正丁醇：醋酸：水（,4,：,1,：,5,，上层）,氯仿：甲醇：乙酸乙酯：水,（,2,：,2,：,4,：,1,，下层）,纯化方法,反相液相色谱（,图谱,）,对于酸性皂苷（尤其是连有葡萄糖醛酸的皂苷类，需加入少量酸，如,0.1%,磷酸），但要注意最后要脱盐处理。,检测器：最好用通用检测器，如蒸发光散射检测器、示差检测器等；若用紫外检测波长,203207nm,。,纯化方法,纯化方法,制备薄层,(,必要时可二次展开,),尽管三萜皂苷的结构解析较为繁琐，但还是有其规律可循。可从下几个方面加以考虑：,分子量及分子式的确定；,母核类型、糖基个数、种类及苷化位置；,糖基连接位置及顺序；,苷键的构型；,二、结构解析规律,最常用的方法就是红外光谱、质谱及核磁共振谱。,紫外光谱较少用（三萜皂苷不饱和键少）。,此外，化学方法在确定皂苷的结构时也具有不可替代的作用。如,苷元结构的确定也可采用,脱水,、,氧化,、,还原,、,甲基或双键转位,、,乙酰化,、,甲酯化,等化学反应将未知苷元结构转变为已知化合物，然后将其,IR,、,mp.,、,Rf,或其它光谱数据与已知物数据对照的方法推测其结构。也可采用半合成或全合成方法制备相应的合成产物以确证天然产物的结构,（一）,UV,和,IR,UV,用于判断齐墩果烷三萜类化合物双键类型,一个双键，,205250nm,有微弱吸收,-,不饱和羰基（,-C=C-C=O,），,最大吸收,242250nm,C18-H,（,D/E,cis,），,248249nm,C18-H,（,D/E trans,），,242243nm,异环共轭双键，最大吸收在,240,，,250,，,260nm,同环共轭双键，最大吸收在,285nm,IR,用于区别骨架类型,A(13551392cm,-1,)B(12451330cm,-1,),齐墩果烷型,2,个峰,3,个峰,乌苏烷型,3,个峰,3,个峰,四环三萜类,1,个峰,1,个峰,（二）,MS,的应用：,质谱可用于确定分子量及求算分子式。此外，还可由分子离子丢失的离子碎片的,m/z,推,定或复核分子的部分结构。,五环三萜类的共同规律,有环内双键，,RDA,开裂；无环内双键，从,C,环断裂；,有时,RDA,和,C,环开裂同时发生。,1.,饱和三萜化合物,2.,不饱和三萜化合物,（,a,）,是三萜烯的特征碎片,C-17,为,-COOH,、,-,COOMe,、,内酯时，（,a,）,易失去上述基团生成（,c,），（,c,）,强度稍大于或等于（,a,）,C-17,为,CH,2,OAc,时，（,c,）,大于（,a,）,C-17,为,-CH,3,时，（,c,）,是（,a,）,的三分之一,当,C-11,位氧代的,12,的结构时，除,RDA,开裂外，还有麦氏重排,重排必须具备的条件：,适当位置的杂原子（如：,O,）,-,体系（通常一个双键）,可除去的氢（对,C=O,体系的,位）,（三）,1,H-NMR,甲基质子、与氧同碳质子、双键上烯氢质子、糖的端基质子等。,甲基质子,：,0.6251.500,ppm,最高场甲基（,26-CH,3,）,0.775ppm,（,28-CH,2,OH,、,CH,3,或内酯）,最低场甲基,1.131.15ppm,（,27-CH,3,）,其它小于,1.0ppm,烯氢信号：判断双键取代情况,环内双键,H,5,ppm,环外双键,H,5,ppm,同环双烯与异环双烯的比较：,氧取代的氢信号：判断构型,C3-OAc,3-H 4.004.75ppm,3-H 5.005.48ppm,C4-CH,2,OAc,24-H 4.084.30ppm,dd,23-H 3.773.80ppm,dd,若,C-16,位连有羟基则,27,位甲基质子信号由于去屏蔽效应向低场位移，出现在,1.70-1.90,之间；,16,位氢信号则在,5.0-5.50,处出现一单峰，,H-3,及,H-18,信号通常在,3.22-3.88,之间呈,dd,峰。,1,H-NMR,观察氢谱中,4.70-6.33,范围内端基质子及碳谱中,95-110,端基碳的个数，可以确定糖的个数。糖的端基质子多数呈特征性的双峰，少数呈单峰。,(,四,),13,C-NMR,用于判断结构类型、某些取代基位置及构型,13,C-NMR,由低场向高场可以分为七个区域：,1,、羰基碳区：大于,163,2,、烯碳区：,110160,，区别苷元类型,3,、端基碳区：,95110,，糖的个数及类型,4,、结合碳区：,8095,（糖与苷元，糖与糖）,5,、游离羟基碳区：,6580,（苷元羟基，糖上羟基）,6,、,C,6,碳区：,6065,7,、苷元区：,060,季碳数目,齐墩果烷型,6,个季碳,3042ppm,乌苏烷型和羽扇豆型,5,个季碳,3042ppm,羽扇豆有异丙烯基信号,19.3(q),109.2(t),150.6(s),（,1,）,terpene,（,苷元）区，,0,60ppm,甲基信号,齐墩果烷,8,个甲基,8.933.7ppm,低场：,23-CH,3,27.828.7ppm,29-CH,3,32.733.7ppm,判定,C,20,和,C,4,位取代基构型,30-CH,3,氧取代时，,29-CH,3,32.733.7 27.929.7ppm,，最低场甲基信号。,29-CH,3,氧取代时,30CH,3,23.620,23-CH,3,氧取代时，,24-CH,3,15.6 8.913.7,ppm,最高场甲基信号,(2),糖,C,5,-,C,H,2,O,区（,6065ppm,）,三萜皂苷如分子中含有,6,碳糖如葡萄糖或甘露糖时，在糖分子中的,C,5,CH,2,OH,，,C,6,位数目可辨认，有几个峰就有几个未被取代的糖，如果,6,位被取代时,-CH,2,O-R,则向低场位移至,67,69ppm,。,（,3,）,游离,-OH,区（,6580ppm,）,此区为苷元上的羟基,C,和糖上未被取代的游离,OH,碳,通常母核上连有,羟基,时,碳向低场位移约,34-50,，,碳向低场位移约,2-10,，,碳则向高场位移约,0,-9,如,23,位甲基连有羟基时，碳信号向低场位移至约,64.0,处，,24,位甲基则向高场位移约,13.0,；,16,位连有,羟基时，,C-16,信号低场位移至,74.7,，,C-15,及,C-17,分别向低场位移约,8.2,、,2.1,；连有,羟基时，,C-16,信号向低场位移至,68.0,，其余碳信号与,羟基取代时相近，据此可推断母核上的羟基取代。,对于母核上取代基的构型还可以通过,NOESY,谱解决，通常甲基上质子与空间上相近的质子有较强的相关峰。如从头序楤木中分离得到的以,16-,羟基齐墩果酸为苷元的皂苷中，在,NOESY,谱中，可观测到,16-H,与,27-CH,3,和,29-CH,3,的相关峰。因此推测,16,位羟基为,羟基。,（,4,）结合,C,区（,80,95ppm,）,查看结合,C,区，若,ppm,84,，,则苷元,C,3,OH,一般接有糖，而且糖与糖如果为,1-3,相连的（,G1c-G1c,），,一般在结合碳区则有峰。,（,5,）,anomeric,区（,95,110,）,联糖数目的确定,主要查看,anomeric,区有几条线或几个碳就连有几个糖。,联糖的种类的确定,a.,葡萄糖：可以查看,60,65,区域,C,数目辨认。,b.Ara(f):108109.8ppm,；,Ara(p,):104.5104.9ppm,c.,xyl,:104,106,d.,Rham,101.6,e.,GlcA,：,106,左右,f.Glc,：,105,左右，成酯苷,95,左右,(,如齐墩果酸,28-,葡萄糖苷）。,糖与苷元连接位置的确定,主要根据苷化位移规律，再查看结合,C,区（,8095,）,ppm,84,，,C3-OH,一定接糖；,目前二维核磁共振技术（,COSY,、,HMQC,、,HMBC,、,TOCSY,等）在皂苷结构研究中具有非常重要的作用，通过观察各个信号之间的远程相关，可以找到结构联系的重要信息。,糖与糖的连接式样,（例葡萄双糖的位移效应）,（,6,）烯碳区（,120163nnm,）,根据烯碳区，再结合三萜苷元区（,0,60ppm,），可以区别三萜皂苷的类型如下,问题一,.,分子量及分子式的确定,质谱可用于确定分子量及求算分子式。此外，还可由分子离子丢失的离子碎片的,m/z,推定或复核分子的部分结构。,EI-MS,只能测到皂苷的碎片峰，故应用较少。,FD-MS,、,FAB-MS,、,ESI-MS,在皂苷的解析过程中常被使用。这些质谱均可看到明显的分子离子峰,(,或,M+H,+,、,M+Na,+,等伪分子离子峰,),及依次失去糖碎片的离子峰；,FAB-MS,和,ESI-MS,还可以给出相应的阴离子质谱。,对于新化合物可结合元素分析或,HI-MS,确定分子式。,ESI-MS of,Congmunoside,IX,随着,NMR,技术的发展及各种分离手段的不断完善，大量的三萜皂苷类成分得以被发现并得到确证。仅以五环三萜皂苷为例，到,94,年为止，从天然产物中分得的皂苷就达,400,余种，苷元近,40,余种。,主要参考文献,：,分析化学手册（第五分册），化学工业出版社。,文献（如,Phytochemistry,等）,不定期的综述。,问题二,.,苷元类型及糖种类数目的确定,（一）、羊毛脂烷型（,lanostane,),A/B,、,B/C,、,C/D,环均为反式,C-10,、,C-13,均有,-CH,3,；,C-14,有,-CH,3,；,C-17,为,-,侧链；,C-20,为,R,构型（,-H,）,一般,C-3,位均有,-OH,，,或游离，或成苷，或氧取代。,四环三萜,tetracyclic,triterpenoids,（二）、达玛烷型,(,dammarane,),环与环之间均为反式；,与羊毛脂烷区别在,C-8,及,C-13,位；,C-8,为,-CH,3,，,C-13,为,-H,；,C-10,有,-CH,3,，,C-14,为,-CH,3,，,C-17,为,-,侧链；,C-20,为,R,或,S,构型。,缓和水解（如：,50%HOAc,于,70 C,加热,4,小时，,20,位苷键先断裂，进一步水解，可使,3,位苷键断裂。,（三）、甘遂烷型（,tirucallane,),环与环之间均为反式,与羊毛脂烷型不同之处在于，,13,、,14,为甲基与其相反，分别为,，,-CH3,，,17,位有侧链，,20S,。,（四）、葫芦烷型,(,cucurbitane,),与羊毛脂烷的区别只在,C-9,、,C-10,位；既,C-9,为,-CH3,、,C-10,为,-H,。,（五）、环阿尔廷型,(,cycloartane,),与羊毛脂烷的区别只在,C-9,、,C-10,位；既,C-9,为,-CH3,、,C-10,为,-H,。,五环三萜,（,pentacyclic,triterpenoids,),（,一）、齐墩果烷型,(,oleanane,-,香树脂烷型）,C-3,有,-OH,取代；,C-28-CH,3,易被氧化成酸或,CH,2,OH,、,COOH,。,（二）、乌苏烷型,（,ursane,-,香树脂烷型）,与齐墩果烷的区别：,29-CH,3,在,19,位。,（三）、羽扇豆烷,(,lupane,),E,环为,5,元环；,C-19,位为,-,构型异丙基。,所有环,/,环之间均为反式。,68,种人参皂苷的,8,种皂苷元解析,皂苷的水解,是研究其结构的最重要的反应。最常用的水解方式是酸水解。酸水解多用不同浓度盐酸或硫酸的水溶液或醇溶液，对于不稳定的苷元可用,Smith,降解法或酶解法得到完整的苷元，然后用,TLC,检出单糖的种类，经显色后用薄层扫描仪求得糖的比例。也可用气相或液相色谱的方法对单糖定性定量。对于苷元,28,位的酯键，可用碱水解的方法研究所连接的糖链。,问题三,.,糖间连接位置和连接顺序的确定,将糖链全甲基化，然后水解，其中游离羟基的部位就是连接位置。由于此法需要制备各种甲基化单糖，比较烦琐。,成苷时苷元和糖的,碳产生低场,苷化位移,，位移幅度与糖的构型有关，当苷键不存在立体障碍时，糖的端基羟基处于横键,(,-,葡、,-,甘、,-,鼠、,-,阿,),时苷化位移要大于,端基羟基处于竖,键,(-,葡、,-,阿,),，若糖的,2,位羟基也处于竖键,(-,甘、,-,鼠,),时，苷化位移值更小。,结合解析实践掌握一些常见糖的连接的经验数据是有益的。例如，在葡萄糖的多种连接组合中，糖的,12-,连接使连接点处碳信号向低场位移至,84,左右，,13-,连接出现,89,，,14,连接出现,82,12-13-,连接出现,88,79,(13)-(13)-12-,连接出现,79,88,88,(13)-(13)-,连接出现,88,88,，熟悉这些化学位移将有利于糖连接的初步判断。,早期决定糖链,连接顺序,的方法主要用缓和水解法，即用稀酸水解、酶解、乙酰解、碱水解等方法，将糖链水解成小的片段，然后分析这些片段的连接顺序。质谱可出现皂苷由糖链末端依次失去糖的一系列信号，对判断糖间连接顺序有很大帮助。,目前二维核磁共振技术（,COSY,、,HMQC,、,HMBC,、,TOCSY,等）在皂苷结构研究中具有非常重要的作用，通过观察各个信号之间的远程相关，可以找到结构联系的重要信息。如,COSY,谱可以归属相邻,H,信号，,TOCSY,谱可以将自旋系统内一组,H,信号明确地归属；再借助,QC,谱可以将与之相对应的,C,信号加以归属；,HMBC,谱对于判定成苷位置具有重要意义。,问题四,.,苷键构型,糖的,H-1,和,H-2,的偶合常数,J,H1-H2,可用于决定端基碳构型,(,甘露糖及鼠李糖除外,),。,J,值在,6-8,之间为,型；,J,值在,3-4,之间为,型。,利用门控去偶技术得到的端基碳和端基质子之间的偶合常数，,1,J,C1-H1,也可以区分苷键的构型，当端基氢处于平伏键时,(,型,),，,1,J,C,1,-H,1,值为,169-171Hz,；,当端基氢处于直立键时,(,型,),，,1,J,C,1,-H,1,值为,158-162Hz,。,比旋光度方法也可确定苷键构型,。,利用酶解法理论上也可以确定苷键构型；此外，红外光谱中,840 cm,-1,是,-L-,吡喃糖苷，,890 cm,-1,则是,-D-,吡喃糖苷的特征吸收峰。,13,C-NMR,法,对于糖的绝对构型，作者在实验中参照,Hara,等所用的方法，将皂苷酸水解后用,Amberlite,MB-3,脱盐，然后以盐酸,L-,半胱氨酸甲基酯处理，最后用,hexamethyl,disilazane-trimethyl,chlorosilane,(HMDS-TMCS),三甲基硅烷化，以,GC-MS,分析所得产物，通过对比水解得到的单糖与标准糖的衍生物的保留时间及裂解碎片证实了糖的种类及构型。,对人参皂苷而言，,C,20,位,OH,的位置决定,C,20,位立体构型（,R,，,S,），,C,20,位一对差向异构体的化学位移不同。,R,型与,S,型之间差别如下：,20(R)-20(S)13C 16C 17C 20C 21C 22C,+0.84 -0.2 -3.7 +0.6 -5.0 +7.5,X-,射线单晶衍射法,1H-NMR,的,Mosher,法,该方法是将仲醇（或伯胺）分别与（,R,）和（,S,）,-,甲氧基,-,三氟甲基,-,苯基乙酸（,MTPA,）反应形成酯（,Mosher,酯），然后比较（,R,）和（,S,）,-MTPA,酯的,1H-NMR,得到,（,=,S-R,），在与,Mosher,酯的构型关系模式图比较的基础上，根据,的符号来判断仲醇手性碳的绝对构型。,辽东,楤木,Aralia,elata,(,Miq,.)Seem.,三,.,结构解析实例,Chart 2,aglycone,of,saponins,from,Aralia,I,II,III,IV,Structure elucidation of new compounds,1.,Oleanolic,acid type,saponins,C,59,H,93,O,28,Congmunoside,XI,1,H-NMR SPECTRUM,13,C-NMR SPECTRUM,C-3,C-28,1,H-,1,H COSY SPECTRUM,6.22,6.10,5.73,5.00,4.92,TOCSY SPECTRUM,HMQC SPECTRUM,A-1,G-1,G-1,G-1,GlcA-1,106.4,105.3,105.5,107.9,95.7,HMQC SPECTRUM,G-H-6,GA-H-3,A-H-3,GA-H-4,A-H-2,A-H-4,HMBC SPECTRUM,Positive ESI-MS,Negative ESI-MS,Table2.NMR date of Compound 5,1014,16,(in pyridine),C-position,5,10,11,12,13,14,16,1,38.7,38.6,38.5,38.7,38.7,38.7,38.4,2,26.5,26.4,26.5,26.7,26.1,26.6,26.4,3,89.3,89.3,89.3,89.4,89.4,89.8,89.2,4,39.8,39.5,39.5,42.1,39.8,39.6,39.7,5,55.7,55.7,55.6,55.9,55.9,55.8,55.5,6,18.5,18.4,18.4,18.5,18.5,18.5,18.3,7,33.9,33.1,33.1,33.1,33.1,32.5,32.9,8,39.5,39.8,39.8,39.7,39.7,39.9,39.3,9,47.9,47.9,47.9,47.9,47.9,48.0,47.8,10,36.9,36.8,36.9,36.9,36.9,36.9,36.7,11,23.7,23.7,23.6,23.4,23.3,23.4,23.5,12,122.1,122.8,122.9,123.2,123.2,122.7,122.6,13,144.1,144.1,144.1,144.2,144.1,144.0,143.9,14,42.1,41.7,42.1,42.1,42.1,42.1,41.9,15,28.2,28.2,28.2,28.2,28.2,28.2,28.1,16,23.4,23.4,23.3,23.6,23.6,23.6,23.2,17,46.9,46.9,46.9,46.9,46.9,46.9,46.8,18,41.7,42.1,41.6,41.6,41.6,41.7,41.5,19,46.2,46.2,46.2,46.2,46.2,46.2,46.0,20,30.7,30.7,30.7,30.7,30.7,30.7,30.6,21,33.9,33.9,33.9,33.9,33.9,34.0,33.8,22,32.5,32.5,32.5,32.5,32.5,33.1,32.3,23,27.9,27.9,27.9,27.8,27.8,27.6,27.8,24,16.7,16.7,16.9,16.5,16.4,16.3,16.8,25,15.5,15.5,15.5,15.6,15.6,15.3,15.4,26,17.4,17.4,17.4,17.3,17.4,17.4,17.3,27,26.0,26.0,26.1,26.1,26.1,26.1,25.9,28,175.9,176.4,176.5,176.4,176.4,176.4,176.3,29,33.2,33.9,33.1,33.1,33.1,33.1,32.9,30,23.6,23.4,23.6,23.5,23.4,23.8,23.4,3-O-Sugar,GlcA,Ara,Ara,GlcA,1,104.9,104.8,106.4,105.3,105.4,105.3,106.3,2,79.2,78.9,76.2,77.3,77.0,78.8,75.7,3,88.6,88.4,82.2,84.1,84.7,87.6,82.0,4,69.9,69.8,73.8,69.6,69.6,71.5,73.7,5,77.8,77.6,75.7,66.9,66.9,78.8,76.1,6,63.3,61.9,172.6,169.3,172.6,Glc,1,103.8,103.5,105.5,104.7,104.7,65.1,105.3,2,76.4,76.4,75.7,76.1,76.2,30.8,75.5,3,78.7,78.8,78.3,79.1,78.6,19.2,78.2,4,71.5,72.4,70.9,71.4,71.3,13.7,71.4,5,77.7,77.9,78.4,78.2,77.8,78.2,6,62.1,62.1,62.6,69.8,69.9,69.7,Glc,Ara,Xyl,1,104.6,103.9,107.9,105.5,105.6,104.8,105.4,2,75.4,74.1,82.0,75.1,75.4,76.1,74.9,3,78.6,87.6,79.3,78.3,77.9,75.3,78.1,4,72.5,69.9,87.5,71.8,71.5,71.4,71.3,5,78.6,78.3,62.1,78.4,78.4,67.2,78.4,6,62.2,62.3,62.4,62.3,62.1,Gal,105.1,104.7,105.2,107.8,75.3,74.0,72.9,81.9,76.2,87.9,77.3,79.2,71.5,69.5,70.1,87.3,78.3,78.5,76.5,61.9,62.3,62.2,61.9,105.5,75.2,76.9,71.5,77.9,61.9,C-28-Glc,Glc,Glc,Glc,Glc,1,95.9,105.1,95.6,95.6,95.6,95.8,95.4,2,74.1,75.0,75.8,75.8,73.9,74.2,75.5,3,78.8,78.4,78.4,78.4,78.9,78.9,78.4,4,71.1,71.4,70.7,70.7,70.8,71.1,70.6,5,79.3,77.9,77.9,77.9,78.2,79.3,77.7,6,62.5,62.4,69.3,69.3,69.4,62.2,69.1,95.7,105.3,105.3,105.2,105.0,74.1,75.1,75.1,74.9,74.7,78.8,78.6,78.6,78.4,78.1,70.1,71.5,71.5,71.2,71.3,79.3,78.7,78.7,78.2,78.2,63.3,62.3,62.3,62.5,62.4,2.,hederagenin,type,saponins,R1 R2,7 -Glc2-Glc -,Glc,3,Glc,9 -Ara2-Glc -,Glc,3,Glc,23 -Glc2-Glc -,Glc,3,Glc3-Glc,Congmunoside,IX,C,53,H,86,O,23,1,H-NMR,SPECTRUM,13,C-NMR SPECTRUM,C-28,C-3,C-23,C-24,1,H-,1,H COSY SPECTRUM,6.29,4.99,5.39,5.22,TOCSY SPECTRUM,6.29,5.39,5.22,4.99,HMQC SPECTRUM,HMQC SPECTRUM,104.8,104.3,104.2,97.8,A-H-3,A-H-2,A-H-5,A-H-4,A-H-5,HMBC SPECTRUM,HMBC SPECTRUM,C-3,Ara-1,Glc-1,A-3,Glc-1,A-2,ESI-MS,Table 3 NMR date of 7,，,9,23,No,7,9,23,1,38.7,38.7,38.7,2,26.0,25.9,25.9,3,83.0,82.4,83.0,4,42.1,43.5,43.4,5,48.1,47.8,47.9,6,18.2,18.1,18.2,7,33.1,32.7,32.8,8,39.9,39.8,39.9,9,47.9,48.1,48.0,10,36.9,36.8,36.8,11,23.8,23.8,23.8,12,122.8,122.8,122.8,13,144.1,144.0,144.1,14,41.7,42.1,42.1,15,28.2,28.2,28.2,16,23.6,23.3,23.6,17,46.1,46.9,46.1,18,43.5,41.6,41.7,19,46.9,46.1,46.9,20,30.7,30.7,30.6,21,33.9,33.9,33.9,22,32.8,32.5,32.5,23,64.8,64.6,64.0,24,13.3,13.3,13.2,25,16.0,16.1,15.9,26,17.5,17.5,17.4,27,25.9,25.9,25.9,28,176.4,176.4,176.3,29,32.5,33.0,32.9,30,23.4,23.6,23.4,3-O-Glc,Ara,1,103.9,104.2,103.6,2,79.3,77.3,79.1,3,88.9,83.4,88.7,4,69.8,68.6,71.2,5,77.6,65.8,77.5,6,62.3,62.3,Glc,1”,103.7,104.3,103.4,2”,76.3,75.9,76.2,3”,78.6,77.5,77.9,4”,71.5,71.9,71.4,5”,77.7,78.6,78.4,6”,62.1,62.2,62.3,Glc,1”,104.8,104.8,103.9,2”,75.3,75.9,74.0,3”,78.6,78.2,87.6,4”,72.2,71.4,69.8,5”,78.6,78.4,77.5,6”,62.4,62.9,62.3,Glc,105.0,74.9,78.4,69.8,77.9,61.9,C-28-Glc,1,95.7,97.8,95.6,2,74.1,74.1,74.0,3,78.8,78.8,78.8,4,71.1,71.0,72.1,5,79.2,79.3,79.1,6,63.1,62.1,63.1,3.,echinocystic,acid type,saponins,R1 R2,1,-Ara,3,-Glc -,Glc,3,-Ara,3,-Glc,3,-Glc -,Glc,4,-Glc,3,-Glc,3,-Glc -,Glc,6,-Glc,2,-Glc -,Glc,3,Glc,15,-Glc,2,-Glc -H,3,Glc,3,-Glc,18,-Glc,2,-Glc -H,3,Glc,19,-Glc,4,-Glc -H,20,-Glc,3,-Glc -H,24 -Glc,3,-Glc,3,-Glc -H,Congmunoside,I,C,53,H,86,O,23,1,H-NMR SPECTRUM,27-H,16-H,13,C-NMR SPECTRUM,107.3,106.2,95.7,74.1,49.1,35.9,C-3,C-28,1,H-,1,H COSY SPECTRUM,NOESY SPECTRUM,26-Me,16-H,TOCSY SPECTRUM,4.73,6.33,5.39,HMQC SPECTRUM,A-H-3,C-16,HMBC SPECTRUM,ESI-MS,Table 1.,13,C-NMR Spectrum Data of Compounds,1,，,3,，,4,，,6,，,15,18,19,20,24,in C,5,D,5,N(500MHz),C-position,1,3,4,6,15,18,19,20,24,1,38.9,38.8,38.8,38.8,38.7,38.8,38.8,38.8,38.8,2,26.7,27.1,27.2,26.5,26.5,26.6,26.6,26.5,26.6,3,88.5,88.7,88.9,89.3,89.3,89.4,89.0,89.1,89.0,4,39.6,39.6,39.5,39.5,39.5,39.6,39.6,39.5,39.6,5,55.9,55.9,55.8,55.9,55.8,56.0,56.0,55.9,55.9,6,18.5,18.5,18.5,18.5,18.5,18.6,18.6,18.5,18.6,7,33.4,33.2,33.2,33.4,33.4,32.9,33.6,33.5,33.4,8,40.1,40.1,40.0,40.7,39.8,40.0,40.0,39.9,39.9,9,47.2,49.1,47.1,47.1,47.1,47.2,47.2,47.2,47.2,10,37.1,37.0,36.9,36.9,36.9,37.0,37.1,37.0,37.1,11,23.8,23.8,23.8,23.8,23.8,23.9,23.9,23.8,23.9,12,122.7,122.7,122.7,122.7,123.2,122.4,122.4,122.3,122.4,13,144.4,144.4,144.4,144.2,145.1,145.2,145.2,145.1,145.2,14,42.1,42.1,42.0,42.0,42.1,42.2,42.2,42.1,42.2,15,35.9,36.1,36.1,36.1,36.1,36.3,36.2,36.2,a),36.2,16,74.1,74.5,74.5,74.1,74.7,74.8,74.8,74.7,74.6,17,49.1,49.1,49.1,49.1,48.8,49.0,49.0,48.9,48.9,18,41.3,41.2,41.3,41.3,41.4,41.5,41.5,41.4,41.5,19,47.2,47.1,47.5,47.1,47.3,47.4,47.2,47.3,47.4,20,30.8,30.8,30.8,30.8,31.0,31.1,31.1,31.0,31.1,21,36.1,33.4,36.9,35.9,32.9,33.4,36.2,36.1,a),36.2,22,32.2,32.2,32.1,32.2,33.1,33.4,32.9,33.2,32.9,23,28.1,28.0,28.1,27.9,27.9,28.1,28.3,28.1,28.2,24,16.9,16.9,17.0,16.7,16.8,16.8,17.1,17.0,17.1,25,15.7,15.7,15.6,15.6,15.5,15.6,15.6,15.5,15.6,26,17.6,17.5,17.5,17.5,17.4,17.5,17.5,17.4,17.6,27,27.2,27.2,27.2,27.2,27.2,27.3,27.3,27.2,27.3,28,175.9,175.9,175.9,175.9,180.0,180.0,180.0,179.8,180.1,29,33.2,33.2,33.2,33.2,33.1,33.4,33.4,32.7,33.4,30,24.6,24.5,24.5,24.6,23.6,24.8,24.8,24.8,24.8,3-O-Sugar,Ara,Ara,Glc,1,107.3,107.4,106.3,104.9,104.8,105.0,106.5,106.2,106.4,2,71.8,71.9,74.5,79.3,79.0,79.4,75.4,75.4,74.8,3,84.0,83.3,88.2,88.6,88.4,88.8,76.3,89.0,88.3,4,69.2,69.4,69.8,69.9,69.9,70.1,81.7,69.9,69.8,5,66.9,67.1,78.2,77.6,77.7,77.8,77.0,77.8,78.3,6,62.2,63.3,61.9,63.5,62.5,62.5,a),62.2,Glc,1,106.2,105.7,105.8,103.8,103.6,103.9,105.0,105.9,105.8,2,75.6,74.3,74.4,76.4,76.3,76.5,74.9,74.4,74.4,3,78.3,88.3,88.4,78.6,78.3,78.7,78.3,78.2,88.5,4,71.5,69.6,69.8,71.0,71.4,71.7,71.6,71.6,69.8,5,78.6,78.2,78.1,77.9,77.7,77.7,78.5,78.6,78.3,6,62.2,62.1,62.2,62.1,63.3,62.4,62.5,62.6,a),62.6,Glc,1,105.9,105.2,104.6,104.0,104.8,105.3,2,75.6,75.6,75.4,74.1,75.5,75.7,3,78.1,78.7,78.8,87.6,78.7,78.7,4,71.5,71.6,72.5,69.8,72.6,71.7,5,79.4,77.8,78.5,77.6,78.7,77.9,6,62.2,62.5,62.2,62.4,62.6,62.6,C-28-Glc,1,95.7,95.8,95.9,95.3,105.1,2,74.4,74.0,74.3,74.3,75.1,3,78.8,78.3,78.9,79.1,77.9,4,71.1,70.9,71.1,71.0,72.1,5,79.3,78.1,79.3,79.3,78.5,6,62.6,62.5,62.5,62.5,62.5,4.,caulophyllogenin,type,saponins,R1 R2,2 -Ara3-Glc -,Glc,8 -Glc2-Glc -,Glc,3,Glc3-Glc,17 -Ara3-Glc -H,21 -Glc2-Glc -H,3,Glc3,Congmunoside,II,C,53,H,86,O,23,1,H-NMR SPECTRUM,27-H,13,C-NMR SPECTRUM,175.9,106.5,106.3,95.7,64.1,81.9,1,H-,1,H COSY SPECTRUM,6.31,5.29,4.93,TOCSY SPECTRUM,1,H-,1,H COSY SPECTRUM,6.31,5.29,4.93,TOCSY SPECTRUM,HMQC SPECTRUM,95.7,106.3,106.5,H-16,A-H-3,84.1,HMBC SPECTRUM,C-3,A-3,ESI-MS,Table 4.,13,C-NMR Spectrum Data of Compounds,2,8,17,21,in C,5,D,5,N(500MHz),C-position,2,8,17,21,1,38.8,38.7,38.9,38.7,2,26.2,25.9,26.3,25.9,3,81.9,82.9,81.9,83.2,4,43.5,43.4,43.6,4