1、L/O/G/OZhangfan Three new steroidal saponins fromFritillaria pallidifloraContentsIntroduction 1Experimental2Results3 Discussion4Fritillaria pallidiflora(贝母贝母)Fritillaria pallidiflora Schrenk is one of the well-known traditional Chinese medicines.It belongs to the Fritillaria genus of Liliaceae famil
2、y(百合科贝母百合科贝母属属)and is mainly distributed in Xinjiang Province of China.The dry bulbs of F.pallidiflora have been used as antitussive(止咳药止咳药),antiasthmatic(平喘药平喘药),and expectorant(祛痰药祛痰药)agents for hundreds of years.Introduction The alkaloids（生物碱）（生物碱）and saponins（皂苷）（皂苷）are the main effective ingred
3、ients（有效成分）（有效成分）in the Fritillaria genus of Liliaceae family.However,a literature survey concerning the secondary metabolites（次生代谢）（次生代谢）of F.pallidiflora showed that no systematic chemical work on the saponin constituents of the plant had been carried out.Therefore,a phytochemical investigation（植物
4、化（植物化学研究）学研究）on the saponin constituents of the dry bulbs of F.pallidiflora was carried out.The structures of the isolated compounds were elucidated by UV(紫外紫外),IR（红外）（红外）1H NMR and 13C NMR（核磁共振）（核磁共振）analysis.Plant material The dry bulbs of F.pallidiflora Schrenk(2.0 kg)were collected in Yining Xin
5、jiang Province of China in June 2008,and identified by Dr Yong Tan of Shihezi University.Main Equipments HPLC(高效液相色谱仪高效液相色谱仪):Waters 2695 Separations NMR(核磁共振光谱仪核磁共振光谱仪):BRUKER-APX-600 Rotary evaporation(旋转蒸发仪旋转蒸发仪):EYBLA Recirculated water(循环水式真空泵循环水式真空泵):SHZ-D9 Methods Extraction（提取）（提取）Isolation
6、(分离分离)Technology roadmap70%EtOHThe dry bulbsextractsreduced pressurebrown residue(98 g)suspendedn-BuOHsilica gel elutingHPLCUV,IR,NMR,MSNew compounds70%EtOHextractsreduced pressure70%EtOHextractsbrown residue(98 g)reduced pressureHextractssuspendedbrown residue(98 g)reduced pressureextractsn-BuOHsus
7、pendedbrown residue(98 g)reduced pressureextractssilica gel elutingn-BuOHsuspendedbrown residue(98 g)reduced pressureextractsHPLCsilica gel elutingn-BuOHsuspendedbrown residue(98 g)reduced pressureextractsUV,IR,NMR,MSHPLCsilica gel elutingn-BuOHsuspendedbrown residue(98 g)reduced pressureextractsNew
8、 compoundsUV,IR,NMR,MSHPLCsilica gel elutingn-BuOHsuspendedbrown residuereduced pressure70%EtOHextracts减压浓缩减压浓缩硅胶柱洗硅胶柱洗脱脱70%乙醇乙醇正丁正丁醇醇Results Three new steroidal saponins,pallidiflosides A(1),B(2),and C(3)were obtained by spectroscopic techniques and chemical means.Compound 126-O-D-glucopyranosyl-(2
9、5R)-furost-5,20(22)-dien-3,26-diol-3-O-D-xylopyranosyl(1 4)-L-rhamnopyranosyl(1 2)-D-glucopyranoside(1)Discussion Compound 1 was obtained as a white amorphous solid with 20D-56.1(MeOH,c0.052)and gave positive results with LiebermannBurchard and Ehrlich reagent tests.The IR spectrum showed absorption
10、 bands for hydroxyl(3364 cm-1)and a glycosidic linkage(10001100 cm-1).白色无定形固体，浓白色无定形固体，浓硫酸硫酸-醋酐反应阳性醋酐反应阳性红外光谱显示：羟基红外光谱显示：羟基在在3364有吸收带；在有吸收带；在1000-1100有配糖键有配糖键1HNMRspectrum (氢谱氢谱)three methyl singlets(3个个甲基信号甲基信号)one methyl doublet（1个个甲基双峰甲基双峰）one methyl doublet（1个甲基双峰）个甲基双峰）the aglycone moities（糖苷配基
11、（糖苷配基）four anomeric proton signals（4个异端质子信号）个异端质子信号）the sugar moities（糖部分）（糖部分）13C NMRspectrum(碳谱碳谱)the aglycone moities（糖苷配基（糖苷配基）two pairs of olefinic carbons（两对烯碳信号）（两对烯碳信号）the sugar moities（糖部分）（糖部分）four anomeric carbons(4个端个端基碳基碳)of sugars The above evidences indicated that 1 was a furostanol s
12、aponin（呋（呋喃甾烷型皂甙喃甾烷型皂甙 ）with four sugar units.Compound 1Compound 226-O-D-glucopyranosyl-3,26-dihydroxyl-20,22-seco-25(R)-furost-5-en-20,22-dione-3-O-L-rhamnopyranosyl(1 2)-D-glucopyranoside Compound 2 was obtained as a white amorphous solid with 20D-59.2(MeOH,c 0.052)and it gave positive results wit
13、h LiebermannBurchard and positive Ehrlich reagent tests.The IR spectrum showed absorption bands for hydroxyl(3363 cm-1),carbonyl of ester(1745 cm-1),and carbonyl of ketone(1713 cm-1).红外光谱显示：羟基红外光谱显示：羟基在在3363有吸收带；在有吸收带；在1745有酯基。在有酯基。在1713处有酮基处有酮基1HNMRspectrum (氢谱氢谱)the aglycone moities（糖苷配基（糖苷配基）thre
14、e methyl singletsone olefinic proton（1个烯个烯质子信号）质子信号）the sugar moities（糖部分）（糖部分）three anomeric protons（3个异个异端质子信号）端质子信号）13C NMRspectrum(碳谱碳谱)the aglycone moities（糖苷配基（糖苷配基）trisubstituted double bonds（三（三取代双键）取代双键）four methyl groupsa ketone carbonyl（酮羰基）（酮羰基）an ester carbonyl（酯羰基）（酯羰基）13C NMRspectrum(
15、碳谱碳谱)the sugar moities（糖部分）（糖部分）three anomeric carbon signals（三个异头碳信（三个异头碳信号）号）All this indicated that 2 was a furostanol saponin（呋喃甾烷型皂甙（呋喃甾烷型皂甙 ）with three sugar units.Compound 2Compound 3(25R)-spirost-5-ene-3,17-diol-3-O-D-glucopyranosyl(1 4)-D-galactopyranoside Compound 3 was obtained as a white
16、 amorphous solid with 20D-47.2(MeOH,c 0.051)and gave results positive in LiebermannBurchard and negative in Ehrlich reagent tests.The IR spectrum showed absorption bands for hydroxyl(3421 cm-1)and a glycosidic linkage (10001100 cm-1).红外光谱显示：羟基红外光谱显示：羟基在在3421有吸收带；在有吸收带；在1000-1100处糖苷键处糖苷键1HNMRspectrum
17、 (氢谱氢谱)the aglycone moities（糖苷配基（糖苷配基）two methyl singletstwo methyl doubletsone olefinic proton（1个烯个烯质子信号）质子信号）the sugar moities（糖部分）（糖部分）two anomeric proton signals（2个异端质子信号）个异端质子信号）13C NMRspectrum(碳谱碳谱)the aglycone moities（糖苷配基（糖苷配基）one quaternary carbon（4碳信碳信号）号）signaltwo olefinic carbon signals（
18、两个烯碳信号）两个烯碳信号）the sugar moities（糖部分）（糖部分）two anomeric carbons（异头异头碳）碳）of sugars The above evidences indicated that 3 was a spirostanol saponin（螺螺旋甾烷醇旋甾烷醇）with two sugar units.Compound 3 As a result,three new steroidal saponins（甾体（甾体皂苷皂苷）,pallidiflosides A(1),B(2),and C(3)were obtained.All these results were in good agreement with the literature data ConclusionAcknowledgment This work was financially supported by National Key Technologies R&D Program of China during the 11th Five-Year Plan Period(2009ZX09501-011)and National Major Special Science and Technology Project(2010ZX09401-304-104A).
