CN103772467B - A kind of saponin(e isomer separation purification process - Google Patents
A kind of saponin(e isomer separation purification process Download PDFInfo
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Abstract
The invention provides a kind of saponin(e isomer separation purification process.Efficiently separating for saponin(e isomers is realized under alcohol water elution system using hydrophilic Interaction Chromatography.Flowing phase composition is methyl alcohol, ethanol, isopropanol, n-butanol or water, without buffer salt addition, is easy to sample preparation to post-process.Using linear gradient, stepwise gradient or isocratic elution mode.Sanchi leaf water extract reverse-phase chromatography preparative separation 4 cuts of gained are chosen, 4 saponins isomers is prepared by hydrophilic chromatographic, totally 12 saponin(es, including 1 new saponin(e and 2 saponin(es got from sanchi leaf first.The method can realize the efficient preparation of saponin(e isomers, and particularly to the saponin(e isomers containing arabino-furanosyl, pyranoid form aralino or pyranoid form xylosyl, the method shows excellent separating effect.It is that the activity research of saponins compound and the new drug development of single component provide material base such that it is able to saponin(e storehouse of enriching constantly.
Description
Technical field
It is specifically a kind of to utilize hydrophilic Interaction Chromatography in alcohol water the present invention relates to isolating and purifying for saponins compound
The method that saponin(e isomers is efficiently separated in eluent system.The method is to Arabic comprising arabino-furanosyl, pyranoid form
The saponin(e isomers of glycosyl or pyranoid form xylosyl shows preferably separating effect.
Background technology
Saponin(e（Saponins）It is aglycon（Sapogenins）It is triterpene（Triterpenoids）Or spirostane class
（Spirostane）One class glucosides of compound.In the chemical constitution of saponin(e, because aglycon has different degrees of lipophilicity,
Sugar chain has stronger hydrophily, saponin(e is turned into a kind of surfactant, and firmly shaking its aqueous solution can produce lasting sex vesicle
Foam.
Saponins compound is to be permitted also with different physiological roles in addition to characteristics such as surface-active, haemolysis, malicious fishes
The main active of many Chinese medicines.The bioactivity of saponin(e with aglycon except having outside the Pass, and the structure with sugar chain is also in close relations.
S.G.Sparg, Wang Nan, Zhang Yun peak and Zhang Cunli etc. are reviewed to the bioactivity that saponins compound has respectively, this
A little activity are main to be included preventing and treating disease of cardiovascular system, anticancer, hypoglycemic, reducing blood lipid, liver protection, anti-inflammatory, antiallergy, anti-micro- life
Thing, anti-aging, antifertility etc.（S.G.Sparg,M.E.Light,J.van Staden,Biological activities and
distribution of plant saponins,Journal of Ethnopharmacology94(2004)219-243；King
The progress of nanmu saponin(e bioactivity, medical graduate students journal, 2007,20 (2):211-214；Zhang Yun peak, Wei Dong, Deng Yan
Such as, the bioactivity research new development of triterpenoid saponins, Chinese patent drug, 2006,28 (9) are waited:1349-1353；Zhang Cunli, Wu Zhanku,
Ma Huiling, waits the bioactivity research of steroid saponins to be in progress, Xibei Forest College's journal, 2003,18 (2):95-100.）.
The aglycon of saponin(e and the structure diversity of sugar chain are the architecture basics of the numerous physiologically actives of saponin(e, but this species diversity
Also the structure elucidation to saponin(e brings very big difficulty, particularly saponin(e isomers.Because the nuclear-magnetism characterizing method of routine needs
The high-purity saponin monomer of milligram level.Reverse-phase chromatography is most widely used chromatographic technique in current saponin separation, but for only
The separating power of the saponin(e isomers reverse-phase chromatography that end group sugar has differences is often not enough.For example, 3 kinds of common ginseng soaps
Glycosides（Ginsenosides）Rc,Rb2And Rb3, their difference existed only on end group pentose.Divided using reverse-phase chromatography
From when, even if also being difficult to be efficiently separated using acetonitrile, first alcohol and water Three -dimension flow phase system（X.Qiao et
al.J.Sep.Sci.2011,34,169-175）.Because reverse-phase chromatographic column is not enough to the hydrophilic selectivity of saponin(e.And it is hydrophilic
Action chromatography is but exactly separated using the hydrophilic sex differernce of saponin(e to it, its replacement chromatogram for being expected to turn into reverse-phase chromatography
Pattern separates the saponins isomers.
The content of the invention
The present invention relates to a kind of isolation and purification method of saponin(e isomers.Using hydrophilic Interaction Chromatography in alcohol water mobile phase body
Saponin(e isomers is efficiently separated under system.Flowing phase composition is methyl alcohol, ethanol, isopropanol, n-butanol or water, without buffer salt addition,
It is easy to sample preparation to post-process.Using linear gradient, stepwise gradient or isocratic elution mode.
Wherein described hydrophilic chromatographic post is amphion post（Click XIon, Beijing Hua Puxinchuan Science and Technology Ltd.s）.
Chromatographic run parameter is as follows：Chromatogram column internal diameter is 4.6-100mm；Sample concentration is 1mg/mL-1g/mL；Sample size is 1 μ L-
40mL；Flow velocity is 0.5-480mL/min；Column temperature is 4-60 DEG C.
The flowing phase composition is methanol-water, alcohol-water, isopropanol-water, isopropanol-methanol-water or n-butanol-first
Alcohol-water.
Use the hydrophilic Interaction Chromatography with alcohol water as eluant, eluent that 4 saponins isomeries are prepared from notoginseng leaf extract
Body, including 1 new saponin(e, 2 saponin(es got from sanchi leaf first, totally 12 high-purity saponin(es.The knot of the compound
Structure information is as follows：
The specific preparation method of compound is：
Sanchi leaf water extract is first separated through preparative RPLC, chromatographic condition：Chromatographic column is the post of carbon 18（C18TDE posts）；Water
（A）And acetonitrile（B）Flow visualizing；Gradient is 0-5min, 20% → 32%B of volumetric concentration;5-45min, volumetric concentration 32%
→68%B;45-50min, 68% → 100%B of volumetric concentration;50-55min,100%B；Flow velocity is 300mL/min；Detection wavelength is
203nm；Sample solution concentration is 300mg/mL, and it uses the water dissolves of 20% acetonitrile of volumetric concentration -80%；Sampling volume is 10mL；
1-55min, collection per minute is a, altogether 54 components, and each component is concentrated to dryness rear standby.Component 10 is chosen using hydrophilic
Chromatogram mode is prepared type HPLC separation, chromatographic condition：Chromatographic column is amphion post（Click XIon,150×10mm,
i.d.,10μm）；Water（A）And methyl alcohol（B）Flow visualizing；Volumetric concentration 92%B isocratic elutions；Flow velocity is 2mL/min；Detection ripple
A length of 203nm；Sample size is 50 μ L.Each chromatographic peak is collected, solvent is separately recovered, is tested through nuclear-magnetism and determined, obtain 1,2,3 three
Individual compound.
Choose component 12 and type HPLC separation, chromatographic condition are prepared using hydrophilic chromatographic pattern：Chromatographic column be both sexes from
Sub- post（Click XIon,150×10mm,i.d.,10μm）；Water（A）And methyl alcohol（B）Flow visualizing；Volumetric concentration 95%B etc.
Degree wash-out；Flow velocity is 2mL/min；Detection wavelength is 203nm；Sample size is 50 μ L.Each chromatographic peak is collected, is separately recovered molten
Agent, tests through nuclear-magnetism and determines, obtains 4,5,6 three compounds.
Choose component 17 and type HPLC separation, chromatographic condition are prepared using hydrophilic chromatographic pattern：Chromatographic column be both sexes from
Sub- post（Click XIon,150×10mm,i.d.,10μm）；100% methyl alcohol isocratic elution；Flow velocity is 2mL/min；Detection wavelength
It is 203nm；Sample size is 50 μ L.Each chromatographic peak is collected, solvent is separately recovered, is tested through nuclear-magnetism and determined, obtain 7,8,9 three
Compound.
Choose component 26 and type HPLC separation, chromatographic condition are prepared using hydrophilic chromatographic pattern：Chromatographic column be both sexes from
Sub- post（Click XIon,150×10mm,i.d.,10μm）；100% methyl alcohol isocratic elution；Flow velocity is 1mL/min；Detection wavelength
It is 203nm；Sample size is 50 μ L.Each chromatographic peak is collected, solvent is separately recovered, is tested through nuclear-magnetism and determined, obtain 10,11,12
Three compounds.
The method can realize the efficient preparation of saponin(e isomers, particularly to containing arabino-furanosyl, pyrans
The saponin(e isomers of type aralino or pyranoid form xylosyl, the method shows excellent separating effect.Such that it is able to not
Disconnected abundant saponin(e storehouse, is that the activity research of saponins compound and the new drug development of single component provide material base.
Specific embodiment
In conjunction with example, the present invention will be further described.Example is only limitted to the explanation present invention, rather than to limit of the invention
It is fixed.
1）Sanchi leaf water extract is first separated through preparative RPLC, chromatographic condition：Chromatographic column is the post of carbon 18（C18TDE posts）；
Water（A）And acetonitrile（B）Flow visualizing；Gradient is 0-5min, 20% → 32%B of volumetric concentration;5-45min, volumetric concentration
32%→68%B;45-50min, 68% → 100%B of volumetric concentration;50-55min,100%B；Flow velocity is 300mL/min；Detection wavelength
It is 203nm；Sample solution concentration is 300mg/mL, and it uses the water dissolves of 20% acetonitrile of volumetric concentration -80%；Sampling volume is
10mL；1-55min, collection per minute is a, altogether 54 components, and each component is concentrated to dryness rear standby；Soap is rich in for 4
The component of glycoside isomers（Component 10, component 12, component 17 and component 26）It is prepared using hydrophilic Interaction Chromatography, chromatographic column is
Click XIon posts；Using methanol-water flow visualizing, the chromatogram retention characteristic according to each component, the body of methyl alcohol in mobile phase
Product concentration control is in 80%-100%；Saponin(e isomers is prepared using isocratic elution mode；
Embodiment 1：The preparation of compound 1,2,3
Choose component 10 and type HPLC separation is prepared using hydrophilic chromatographic pattern（Chromatographic column is amphion post；Water（A）
And methyl alcohol（B）Flow visualizing；Volumetric concentration 92%B isocratic elutions）Flow velocity is 2mL/min；Detection wavelength is 203nm；Sample size
It is 50 μ L.Each chromatographic peak is collected, solvent is separately recovered, is tested through nuclear-magnetism and determined, obtain 1,2,3 three compounds.HPLC is examined
Survey purity and be more than 95%, through physical and chemical determination, data are as follows：1, white powder, HR-ESI-MS:[M+H]+,found:
1211.6409;Calcd:1211.6425for C58H99O26.1H NMR(600MHz,pyridine-d5)δ:0.77(1H,s,H-
19),0.92(2H,s,H-18,30),1.08(1H,s,H-29),1.25(1H,s,H-28),1.60(1H,s,H-21),1.62
(1H,s,H-27),1.64(1H,s,H-26),3.28(1H,dd,J=4.2,11.4,H-3),5.29(1H,t,J=7.2,H-24),
4.92(1H,d,J=7.8),5.50(1H,d,J=7.8),5.40(1H,d,J=6.6),5.13(1H,d,J=7.8),4.85(1H,
brs).13CNMR:It is shown in Table 1.2, white powder, HR-ESI-MS:[M+H]+,found:1211.6360;Calcd:
1211.6425forC58H99O26.1H NMR(600MHz,pyridine-d5)δ:0.78(1H,s,H-19),0.93(1H,s,H-
30),0.94(1H,s,H-18),1.09(1H,s,H-29),1.26(1H,s,H-28),1.60(1H,s,H-21),1.62(1H,
s,H-27),1.64(1H,s,H-26),3.28(1H,dd,J=4.2,11.4,H-3),5.31(1H,t,J=6.0,H-24),4.92
(1H,d,J=7.8),5.51(1H,d,J=7.8),5.40(1H,d,J=7.2),5.12(1H,d,J=7.8).13CNMR:It is shown in Table
1.3, white powder, HR-ESI-MS:[M+H]+, found:1211.6390;Calcd:1211.6425for C58H99O26.1H
NMR(600MHz,pyridine-d5)δ:0.78(1H,s,H-19),0.92(1H,s,H-30),0.94(1H,s,H-18),1.09
(1H,s,H-29),1.25(1H,s,H-28),1.59(1H,s,H-21),1.63(2H,s,H-26,27),3.28(1H,dd,J=
4.2,12,H-3),5.29(1H,t,J=7.2,H-24),4.91(1H,d,J=7.8),5.50(1H,d,J=7.8),5.40(1H,
d,J=7.2),5.12(1H,d,J=7.8),4.97(1H,d,J=7.2).13C NMR:It is shown in Table 1.
Embodiment 2：The preparation of compound 4,5,6
Choose component 12 and type HPLC separation is prepared using hydrophilic chromatographic pattern（Chromatographic column is amphion post；Water（A）
And methyl alcohol（B）Flow visualizing；Volumetric concentration 95%B isocratic elutions）Flow velocity is 2mL/min；Detection wavelength is 203nm；Sample size
It is 50 μ L.Each chromatographic peak is collected, solvent is separately recovered, is tested through nuclear-magnetism and determined, obtain 4,5,6 four compounds.HPLC is examined
Survey purity and be more than 95%, through physical and chemical determination, data are as follows：4, white powder, HR-ESI-MS:[M+H]+,found:
1079.5989;Calcd:1079.6002forC53H91O22.1HNMR(600MHz,pyridine-d5)δ:0.78(1H,s,H-
19),0.92(1H,s,H-30),0.93(1H,s,H-18),1.08(1H,s,H-29),1.26(1H,s,H-28),1.60(1H,
s,H-21),1.62(1H,s,H-27),1.64(1H,s,H-26),3.24(1H,dd,J=4.2,11.4,H-3),5.29(1H,t,
J=6.6,H-24),4.90(1H,d,J=7.8),5.35(1H,d,J=7.8),5.12(1H,d,J=7.8),4.85(1H,brs)
.13C NMR:It is shown in Table 1.5, white powder, HR-ESI-MS:[M+H]+, found:1079.6063;Calcd:1079.6002for
C53H91O22.1H NMR(600MHz,pyridine-d5)δ:0.77(1H,s,H-19),0.91(1H,s,H-30),0.93(1H,
s,H-18),1.07(1H,s,H-29),1.24(1H,s,H-28),1.59(1H,s,H-21),1.61(1H,s,H-27),1.63
(1H,s,H-26),3.25(1H,dd,J=4.2,11.4,H-3),5.29(1H,t,J=6.0,H-24),4.89(1H,d,J=
7.2),5.37(1H,d,J=7.8),5.11(1H,d,J=7.8),4.97(1H,d,J=6.0).13CNMR:It is shown in Table 1.6, white powder
End, HR-ESI-MS:[M+H]+,found:1079.5948;Calcd:1079.6002for C53H91O22.1H NMR(600MHz,
pyridine-d5)δ:0.78(1H,s,H-19),0.93(1H,s,H-30),0.95(1H,s,H-18),1.09(1H,s,H-
29),1.26(1H,s,H-28),1.59(1H,s,H-21),1.63(2H,s,H-26,27),3.26(1H,dd,J=4.2,12,H-
3),5.29(1H,t,J=6.6,H-24),4.90(1H,d,J=7.2),5.35(1H,d,J=7.8),5.11(1H,d,J=7.8),
4.97(1H,d,J=7.8).13CNMR:It is shown in Table 1.
Embodiment 3：The preparation of compound 7,8,9
Choose component 17 and type HPLC separation is prepared using hydrophilic chromatographic pattern（Chromatographic column is amphion post；100%
Methyl alcohol isocratic elution）Flow velocity is 2mL/min；Detection wavelength is 203nm；Sample size is 50 μ L.Each chromatographic peak is collected, is returned respectively
Solvent is received, is tested through nuclear-magnetism and determined, obtain 7,8,9 three compounds.HPLC detection purity is more than 95%, through physical and chemical determination, data
It is as follows：7, white powder, HR-ESI-MS:[M+H]+,found:917.5496;Calcd:917.5474for C47H81O17.1H
NMR(600MHz,pyridine-d5)δ:0.78(1H,s,H-19),0.93(1H,s,H-30),0.94(1H,s,H-18),0.97
(1H,s,H-29),1.28(1H,s,H-28),1.59(1H,s,H-21),1.62(1H,s,H-27),1.64(1H,s,H-26),
3.36(1H,dd,J=4.8,12,H-3),5.29(1H,t,J=6.0,H-24),4.92(1H,d,J=7.8),5.12(1H,d,J=
7.8),4.85(1H,brs).13C NMR:It is shown in Table 1.8, white powder, HR-ESI-MS:[M+H]+,found:917.5398;
Calcd:917.5474forC47H81O17.1H NMR(600MHz,pyridine-d5)δ:0.76(1H,s,H-19),0.91(1H,
s,H-30),0.94(1H,s,H-18),0.96(1H,s,H-29),1.27(1H,s,H-28),1.58(1H,s,H-21),1.61
(1H,s,H-27),1.63(1H,s,H-26),3.35(1H,dd,J=4.2,12,H-3),5.29(1H,t,J=6.0,H-24),
4.92(1H,d,J=7.8),5.10(1H,d,J=7.8),4.97(1H,d,J=6.0).13C NMR:It is shown in Table 1.9, white powder,
HR-ESI-MS:[M+H]+,found:917.5402;Calcd:917.5474for C47H81O17.1H NMR(600MHz,
pyridine-d5)δ:0.79(1H,s,H-19),0.93(1H,s,H-30),0.96(1H,s,H-18),0.98(1H,s,H-
29),1.29(1H,s,H-28),1.58(1H,s,H-21),1.63(2H,s,H-26,27),3.36(1H,dd,J=4.8,12,H-
3),5.29(1H,t,J=7.2,H-24),4.92(1H,d,J=7.8),5.11(1H,d,J=7.8),4.97(1H,d,J=7.2)
.13C NMR:It is shown in Table 1.
Embodiment 4：The preparation of compound 10,11,12
Choose component 26 and type HPLC separation, chromatographic condition are prepared using hydrophilic chromatographic pattern：Chromatographic column be both sexes from
Sub- post（Click XIon,150×10mm,i.d.,10μm）；100% methyl alcohol isocratic elution；Flow velocity is 1mL/min；Detection wavelength
It is 203nm；Sample size is 50 μ L.Each chromatographic peak is collected, solvent is separately recovered, is tested through nuclear-magnetism and determined, obtain 10,11,12
Three compounds.HPLC detection purity is more than 95%, and through physical and chemical determination, data are as follows：10, white powder, HR-ESI-MS:[M+
H]+,found:755.4936;Calcd:755.4946forC41H71O12.1H NMR(600MHz,pyridine-d5)δ:0.87
(1H,s,H-19),0.91(1H,s,H-30),0.97(1H,s,H-18),1.01(1H,s,H-29),1.21(1H,s,H-28),
1.60(1H,s,H-21),1.62(1H,s,H-27),1.65(1H,s,H-26),5.30(1H,t,J=7.2,H-24),5.13
(1H,d,J=7.8),4.85(1H,brs).13C NMR:It is shown in Table 1.11, white powder, HR-ESI-MS:[M+H]+,found:
755.4927;Calcd:755.4946forC41H71O12.1H NMR(600MHz,pyridine-d5)δ:0.85(1H,s,H-
19),0.91(1H,s,H-30),0.96(1H,s,H-18),1.01(1H,s,H-29),1.20(1H,s,H-28),1.60(1H,
s,H-21),1.62(1H,s,H-27),1.64(1H,s,H-26),5.30(1H,t,J=7.2,H-24),5.11(1H,d,J=
7.2),4.98(1H,d,J=6.0).13C NMR:It is shown in Table 1.12, white powder, HR-ESI-MS:[M+H]+, found:
755.4964;Calcd:755.4946for C41H71O12.1H NMR(600MHz,pyridine-d5)δ:0.87(1H,s,H-
19),0.94(1H,s,H-30),0.97(1H,s,H-18),1.02(1H,s,H-29),1.21(1H,s,H-28),1.59(1H,
s,H-21),1.64(2H,s,H-26,27),5.30(1H,t,J=7.2,H-24),5.12(1H,d,J=7.8),4.97(1H,d,J
=7.2).13C NMR:It is shown in Table 1.
The compound P1-P12's of table 113C NMR datas (150MHz, deuterated pyridine)（Table The13CNMR(150MHz)
spectral data of the compounds1-12(in pyridine-d5)）
glu:β-D- glucopyranosyls, xyl:β-D- pyranoid form xylosyls, ara:α-L- arabino-furanosyls
Or α-L- pyranoid form aralinos
（glu:β-D-glucopyranosyl,xyl:β-D-xylopyranosyl,araF:α-L-
arabinofuranosyl,araP:α-L-arabinopyranosyl）.
Claims (4)
1. a kind of saponin(e isomer separation purification process, it is characterised in that：Divided from saponin extract using hydrophilic Interaction Chromatography
From purification of high-purity saponin(e isomers；Flowing phase composition be methyl alcohol, ethanol, isopropanol or n-butanol in one or two with
Water, without buffer salt addition, is easy to sample preparation to post-process；Using linear gradient, stepwise gradient or isocratic elution mode, it is described enter
Sample volume is 10mL；1-55min, collection per minute is a, altogether 54 components, and each component is concentrated to dryness rear standby；For 4
Individual component, i.e. component 10, component 12, component 17 and component 26 are prepared using hydrophilic Interaction Chromatography, and chromatographic column is Click
XIon posts.
2. according to saponin separation purification process described in claim 1, it is characterised in that：The flowing phase composition is methanol-water, second
Alcohol-water, isopropanol-water, isopropanol-methanol-water or n-butanol-methanol-water.
3. the preparation method of saponins compound described in a kind of claim 1, it is characterised in that：
1) sanchi leaf water extract is first separated through preparative RPLC, chromatographic condition：Chromatographic column is the post C18TDE of carbon 18；Water (A) and
Acetonitrile (B) flow visualizing；Gradient is 0-5min, 20% → 32%B of volumetric concentration；5-45min, volumetric concentration 32% →
68%B；45-50min, 68% → 100%B of volumetric concentration；50-55min, 100%B；Flow velocity is 300mL/min；Detection wavelength
It is 203nm；Sample solution concentration is 300mg/mL, and it uses the water dissolves of 20% acetonitrile of volumetric concentration -80%；Sampling volume is
10mL；1-55min, collection per minute is a, altogether 54 components, and each component is concentrated to dryness rear standby；Soap is rich in for 4
The component of glycoside isomers is prepared using hydrophilic Interaction Chromatography, and chromatographic column is Click XIon posts；According to the color of each component
Spectrum retention characteristic, the volumetric concentration of methyl alcohol is controlled in 80%-100% in mobile phase；Saponin(e is prepared using isocratic elution mode different
Structure body；
2) choose component 10 and type HPLC separation, chromatographic condition are prepared using hydrophilic chromatographic pattern：Chromatographic column is amphion
Post Click XIon, 150 × 10mm, i.d., 10 μm；Water (A) and methyl alcohol (B) flow visualizing；Volumetric concentration 92%B is isocratic to be washed
It is de-；Flow velocity is 2mL/min；Detection wavelength is 203nm；Sample size is 50 μ L；Each chromatographic peak is collected, solvent is separately recovered, passed through
Nuclear-magnetism experiment determination, obtains three compounds；
Choose component 12 and type HPLC separation, chromatographic condition are prepared using hydrophilic chromatographic pattern：Chromatographic column is amphion post
Click XIon,150×10mm,i.d.,10μm；Water (A) and methyl alcohol (B) flow visualizing；Volumetric concentration 95%B is isocratic to be washed
It is de-；Flow velocity is 2mL/min；Detection wavelength is 203nm；Sample size is 50 μ L；Each chromatographic peak is collected, solvent is separately recovered, passed through
Nuclear-magnetism experiment determination, obtains three compounds；
Choose component 17 and type HPLC separation, chromatographic condition are prepared using hydrophilic chromatographic pattern：Chromatographic column is amphion post
Click XIon,150×10mm,i.d.,10μm；100% methyl alcohol isocratic elution；Flow velocity is 2mL/min；Detection wavelength is
203nm；Sample size is 50 μ L；Each chromatographic peak is collected, solvent is separately recovered, is tested through nuclear-magnetism and determined, obtain three compounds；
Choose component 26 and type HPLC separation, chromatographic condition are prepared using hydrophilic chromatographic pattern：Chromatographic column is amphion post
Click XIon,150×10mm,i.d.,10μm；100% methyl alcohol isocratic elution；Flow velocity is 1mL/min；Detection wavelength is
203nm；Sample size is 50 μ L；Each chromatographic peak is collected, solvent is separately recovered, is tested through nuclear-magnetism and determined, obtain three compounds.
4., according to the preparation method of saponin(e isomers described in claim 3,4 groups of high-purity saponin(es are prepared by hydrophilic chromatographic
Isomers, including 1 unknown saponin(e and 2 saponin(es got from sanchi leaf first, totally 12 saponin(es；
The structural information of the compound is as follows：
Compound 1 is notoginsenoside FP2 (Notoginsenoside FP2), and molecular weight is 1210, and molecular formula is C58H98O26, R1
It is O-glu (2 → 1) glu (2 → 1) xyl, R2It is O-glu (6 → 1) araF；Compound 2 be unknown saponin(e-, molecular weight is
1210, molecular formula is C59H98O26, R1It is O-glu (2 → 1) glu (2 → 1) xyl, R2It is O-glu (6 → 1) araP；Compound 3 is
Notoginsenoside Fc (Notoginsenoside Fc), molecular weight is 1210, and molecular formula is C58H98O26, R1It is O-glu (2 → 1)
Glu (2 → 1) xyl, R2It is O-glu (6 → 1) xyl；Compound 4 is Ginsenoside Rc (Ginsenoside Rc), and molecular weight is
1078, molecular formula is C53H90O22, R1It is O-glu (2 → 1) glu, R2It is O-glu (6 → 1) araF；Compound 5 is ginsenoside
Rb2(Ginsenoside Rb2), molecular weight is 1078, and molecular formula is C53H90O22, R1It is O-glu (2 → 1) glu, R2It is O-glu
(6→1)araP；Compound 6 is ginsenoside Rb3(Ginsenoside Rb3), molecular weight is 1078, and molecular formula is C53H90O22,
R1It is O-glu (2 → 1) glu, R2It is O-glu (6 → 1) xyl；Compound 7 is N-Fe (Notoginsenoside Fe),
Molecular weight is 916, and molecular formula is C47H80O17, R1It is O-glu, R2It is O-glu (6 → 1) araF；Compound 8 is American ginseng saponin
L1 (Quinquenoside-L1), molecular weight is 916, and molecular formula is C47H80O17, R1It is O-glu, R2It is O-glu (6 → 1)
araP；Compound 9 is notoginsenoside Fd (Notoginsenoside Fd), and molecular weight is 916, and molecular formula is C47H80O17, R1For
O-glu, R2It is O-glu (6 → 1) xyl；Compound 10 is ginsenoside Mc (Ginsenoside Mc), and molecular weight is 754, point
Minor is C41H70O12, R1It is OH, R2It is O-glu (6 → 1) araF；Compound 11 is ginsenoside Y (Ginsenoside Y), point
Son amount is 754, and molecular formula is C41H70O12, R1It is OH, R2It is O-glu (6 → 1) araP；Compound 12 is gypenoside XIII
(Gypenoside XIII), molecular weight is 754, and molecular formula is C41H70O12, R1It is OH, R2It is O-glu (6 → 1) xyl.
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