Embodiment
Embodiment 1:
Ginseng contains ginseng self saponin(e enzyme (Y Bao, L An, F Jin(gold phoenix is mediate for C Zhang, H Yu) in the root:
Chem. Pharm. Bull ., 2001,49 (7), 795-798.;
Process BioChem ., 2002,37,793-798.).The present invention extracts the extract (extract of ginseng self enzyme and water-soluble substances) that does not contain saponin(e, contains ginseng self saponin(e enzyme and other water-soluble substanceses from ginseng, with Rb1, Rb2, Rc or Rd monomer or its mixture reaction of protopanoxadiol class, preparation by 20 (
S)-Rg3,20 (
RThe Rg3 saponins that four kinds of saponin(es such as)-Rg3, Rk1, Rg5 form; With GF2 reaction, preparation by 20 (
S)-Rh2,20 (
RThe Rh2 saponins that four kinds of saponin(es such as)-Rh2, Rk2, Rh3 form.
1, the preparation of the extract of the ginseng of ginseng self enzyme and water-soluble substances:
Fresh ginseng 3.5 kilograms (being equivalent to 1 kilogram of trepang) is cut into the sheet of 2 mm thick, adds 8 liters 95% ethanol, extracts ginsenoside 6 hours, separation and Extraction liquid at 50-60 ℃; The extraction using alcohol that adds again 8 liters 70-75% in its slag extracts 3 times altogether; Merge ethanol extract, for the preparation of ginsenoside.Add 8 premium on currency in the slag that ginsenoside extracted, extracting 6 hours below 75 ℃, same method extracts 3 times altogether; United extraction liquid, the product temperature is being evaporated to Baume 60 degree below 60 ℃, centrifugal slagging-off, obtain the extract of 200-300 milliliter ginseng self enzyme and water-soluble substances, is used for the preparation that Rg3 group and Rh2 organize red ginseng saponin.
Also can be with sun-dried dry ginseng: be cut in 1 kilogram of sun-dried dry ginseng of 2 millimeters sheets, add the ethanol of 8 liters 70-75%, extracted ginsenosides 6 hours at 50 ℃; Same method extracts 3 times altogether; Merge ethanol extract, for the preparation of ginsenoside.Add 8 premium on currency in the slag that ginsenoside extracted, extracting 6 hours below 75 ℃, same method extracts 3 times altogether; United extraction liquid, the product temperature is being evaporated to Baume 60 degree below 60 ℃, centrifugal slagging-off, obtain the extract of 200-300 milliliter ginseng self enzyme and water-soluble substances, is used for the preparation that Rg3 group and Rh2 organize red ginseng saponin.
Several times test, raw material can be ginsengs, also can be aquatic foods ginsengs or the trepang of Radix Panacis Quinquefolii or Radix Notoginseng.Add 6-10 times 65-80% ethanol in the ginseng (calculating with dry), extract saponin(e at 50-75 ℃, can repeat 3 times; The 6-10 times of water that adds again dried slag in the slag of extraction saponin(e repeats 2-3 time 60-80 ℃ of extraction; The water-soluble substances that extracts at product temperature concentrating under reduced pressure below 65 ℃, all obtains mixture enzyme and the water-soluble substances extract of needed ginseng.
2, the preparation of Rg3 group red ginseng mixing saponin(e:
Rb1, Rb2, Rc and the Rd mixing saponin(e of the protopanoxadiol class of 50 grams are incorporated in 300 ml waters, grind, above-mentioned 1 ginseng self enzyme and the extract of water-soluble substances and 600 milliliters the water that add again 100 milliliters, 80 ℃ of reactions 6 hours, 110 ℃ went out enzyme 0.5-1 hour, Rb1, the Rb2 of nearly all protopanoxadiol class, Rc and Rd mixing saponin(e be converted into red ginseng saponin 20 (
S)-Rg3,20 (
R)-Rg3, Rk1 and Rg5 mixing saponin(e.Its reaction solution through 1 liter macroporous resin (the HP-20 macroporous resin, MIT produces; Perhaps AB-8 macroporous resin, Tianjin Nankai university produces) post absorption saponin(e, again with 8 liters sugar and the slag that does not adsorb that water-wash away; Use again 6 liters 75-80% ethanol elution saponin(e, its elutriant concentrating under reduced pressure, dry 36 grams 20 (
S)-Rg3,20 (
R)-Rg3, Rk1 and Rg5 mixing saponin(e namely get Rg3 group red ginseng mixing saponin(e.
Several times in the above-mentioned reaction, use Rb1, Rb2, Rc or the Rd monomer saponin of protopanoxadiol class, its result and use Rb1, Rb2, Rc are identical with Rd mixing saponin(e effect; In the preparation, the reaction density 0.1-10% of ginsenoside, the reaction ratio (weight) of the extract of ginsenoside and ginseng self enzyme and water-soluble substances (calculating of ginseng dry product raw material) is 1:0.1-20 times, temperature of reaction 70-120 ℃, all obtains good effect.
Protopanoxadiol class Rb1, Rb2, Rc and Rd monomer or its mix saponin(e and can buy in market.
Embodiment 1 gained Rg3 organizes red ginseng saponin, confirms with high performance liquid chromatography (HPLC) and Ultra Performance Liquid Chromatography-MS (UPLC-Q/TOF).
1) affirmation of Rg3 group red ginseng mixing saponin(e:
The high performance liquid chromatograph that Rg3 group red ginseng saponin is measured, U.S. waters 2695 instruments; Detector, Photodiode Array Detector Waters 2996; Knauer C18 chromatographic column (250mm * 3mm); Moving phase, acetonitrile (A)-water (B); 0 ~ 35min, the degree such as 19%A; 35 ~ 55min, 19%A ~ 29%A linear gradient; 55 ~ 65min, 29%A ~ 40%A linear gradient; 65 ~ 95min, 40%A ~ 100%A linear gradient; Sample size, 10 μ L; Column temperature, 35 ℃; Volumetric flow rate, 0.6mL/min; Detect wavelength, 203nm.
Get above-mentioned Rg3 group and mix 2 milligrams of saponin(es, be dissolved in 1 milliliter the methyl alcohol, through 0.45 μ m membrane filtration, high performance liquid chromatography (HPLC) detected result, with standard substance 20 (
S)-Rg3,20 (
R)-Rg3, Rk1 and Rg5 saponin(e compare, as shown in Figure 1.
As can see from Figure 1, compare with the standard substance saponin(e, Rg3 group contain 20 (
S)-Rg3,20 (
R)-Rg3, Rk1, four kinds of saponin(es of Rg5, total content is more than 90%; Each saponin content be 20 than (peak area ratio) (
S)-Rg3:20 (
R)-Rg3:Rk1:Rg5=29:26:21:24.
2) Ultra Performance Liquid Chromatography-MS (UPLC-Q/TOF) is checked four kinds of saponin(es of Rg3 group:
Ultra Performance Liquid Chromatography-mass spectrometry analyser: UPLC-Q/TOF Premier
TMMicromass (Waters) instrument; Chromatographic column: BEH shield RP18 (2.1 * 100mm, 1.7 μ m, Waters).Ultra Performance Liquid Chromatography-quadrupole rods tandem mass spectrometry coupling detects and finishes at UPLC-Q/TOF PremierTM Micromass (Waters) instrument.The ESI source under the positive and negative ion pattern, is detected under " V " pattern.
Ultra Performance Liquid Chromatography (UPLC) testing conditions: moving phase is (A) 0.1% formic acid-acetonitrile---(B) 0.1% formic acid-water; Gradient elution: 0 ~ 5min, 5%A ~ 95%A linear gradient, 5 ~ 10min, the degree such as 95%A.Sample size, 10 μ L; Flow velocity, 0.5 mL/min; Column temperature, 30 ℃; Detect wavelength, 203nm.
Level Four time flight tandem mass spectrum (Q/TOF) testing conditions: capillary voltage, 2.5kv; Sample well voltage, 35v; Ion source temperature, 100 ℃; Desolventizing temperature degree, 350 ℃; The taper hole gas velocity, 50L/hr; The desolventizing gas velocity, 1000L/hr; The mass scanning scope, 100-1000Da.
In the Rg3 group 20 (
S)-Rg3 and 20 (
R)-Rg3 saponin(e molecular formula is C
42H
72O
13Molecular weight is 784; Rg5 and Rk1 molecular formula, C
42H
70O
12Molecular weight, 766.
Above-mentioned Rg
3The group sample carries out UPLC-Q/TOF and detects, and the collection of illustrative plates that obtains is shown in Fig. 2,3,4.
From Fig. 2,3,4, can find out Rg
3The UPLC collection of illustrative plates of group sample has four obvious ultraviolet absorption peaks, and the HPLC of Fig. 1 detect 20 (
S)-Rg3,20 (
RThe ultraviolet absorption peak of)-Rg3, Rk1, Rg5 is consistent, its retention time is followed successively by 3.77,3.83,4.43,4.49(min).Under the positive and negative ion pattern, detect, obtain the total ion current figure of Rg3 group sample by tandem mass spectrum (Q/TOF), four peaks are also arranged, and it has four obvious ultraviolet absorption peaks consistent with the UPLC collection of illustrative plates, retention time is respectively 3.81,3.87,4.46,4.51(min).
Thus, can carry out the one-level mass spectroscopy at each peak of total ion current, determine the molecular weight of each material in the Rg3 sample.
(1) the one-level mass spectroscopy of the Rg3 group rare saponin(e of ginseng under ESI+ holotype
The quasi-molecular ions of at first Rg3 being organized first, second carries out the one-level mass spectroscopy under ESI+ holotype, the one-level mass spectrum that obtains is shown in Fig. 5,6.
20 (
S)-Rg3 and 20 (
RThe molecular weight of)-Rg3 is 784; From Fig. 5,6, can find out, under ESI+ holotype, make to have added a H on first, second peak material molecule, form m/z and be 785 quasi-molecular ion peak, i.e. [M+H]
+The m/z767 ion is to lose a part H for the m/z785 quasi-molecular ions
2The fragment that O produces; M/z 605 ions are that the m/z785 quasi-molecular ions loses terminal glucosyl group, lose a part H again
2The fragment that O produces; The m/z461 ion is that the m/z785 quasi-molecular ions loses the fragment that two glucosyl groups produce; M/z443, m/z425, m/z407 ion are that the m/z461 fragment loses a part H more successively
2The fragment that O produces; Therefore the molecular weight of first, second peak material is 784; Can be defined as first peak and be 20 (
S)-Rg3; The second peak be 20 (
R)-Rg3.
Again the 3rd, the 4th quasi-molecular ions of Rg3 group carried out the one-level mass spectroscopy under ESI+ holotype, the one-level mass spectrum that obtains is shown in Fig. 7,8.
The molecular weight of Rk1 and Rg5 is 766; From Fig. 7,8, can find out, under ESI+ holotype, make the molecule of the material at the 3rd, the 4th peak add a H, form m/z and be 767 quasi-molecular ion peak, i.e. [M+H]
+The m/z785 quasi-molecular ions belongs to the [M+H that forms under the ESI+ pattern
2O]
+The m/z749 ion is that the m/z767 quasi-molecular ions loses a part H
2The fragment that O produces; M/z 605 ions are that the m/z767 quasi-molecular ions loses the fragment that terminal glucose produces; The m/z587 ion is that the m/z605 quasi-molecular ions loses a part H again
2The fragment that O produces; The m/z443 ion is that the m/z767 quasi-molecular ions loses the fragment that two glucosyl groups produce; M/z425 and m/z407 ion are that the m/z443 fragment loses a part H more successively
2The fragment that O produces.The molecular weight that obtains thus the 3rd, the 4th peak material is 766; Can be defined as Rk1 and Rg5 saponin(e.
(2) Rg
3The rare saponin(e of group ginseng carries out the one-level mass spectroscopy under the ESI-negative mode
At first to Rg
3First, second quasi-molecular ions of group carries out the one-level mass spectroscopy under the ESI-negative mode, the one-level mass spectrum that obtains is shown in Fig. 9,10.
20 (
S)-Rg3,20 (
R)-Rg3 molecular weight is 784; From Fig. 9,10, can find out, under the ESI-negative mode, form [first, second peak material+HCOOH]
-So quasi-molecular ions, and reduce by a H under the negative ion mode is the quasi-molecular ions m/z 829 that obtains by Q/TOF; Its material is lost product nucleus fragment ion m/z 783 behind the monomethyl.So can judge, contain 20 in the sample (
S)-Rg3 and 20 (
R)-Rg3, its molecular weight are 784.
Again the 3rd, the 4th quasi-molecular ions of Rg3 group carried out the one-level mass spectroscopy under the ESI-negative mode, the one-level mass spectrum that obtains is shown in Figure 11,12.
The molecular weight of Rk1 and Rg5 is 766; From Figure 11,12, can find out, under the ESI-negative mode, form [the 3rd, the 4th peak material+HCOOH]
-Quasi-molecular ions, and reduce by a H under the negative ion mode, so the 3rd, the 4th peak obtains quasi-molecular ions m/z 811, it loses product nucleus fragment ion m/z765 behind the monomethyl.So can judge, contain Rk1 and Rg5 in the sample, its molecular weight is 766.
Proof again thus, contain 20 in the prepared Rg3 group (
S)-Rg3,20 (
RFour kinds of rare saponin(es of red ginseng such as)-Rg3, Rk1 and Rg5; Structural formula is respectively:
Through several times test, in the prepared Rg3 group 20 (
S)-Rg3 and 20 (
RThe scope 20-80% of the content ratio of)-Rg3 in Rg3 group total saponins.
Embodiment 2:
The preparation of Rh2 group red ginseng saponin: react with ginseng self enzyme of embodiment 1 and extract and the GF2 of water-soluble substances, preparation Rh2 group is mixed saponin(e.
The required raw material GF2 of preparation Rh2 saponins is with patent and document (European patent EP 1354944, US Patent No. 7,759,101B2, document H Yu ... F Jin:
Chem Pharm. Bull ., 2007,55,231-235.) middle method prepares, and namely uses ginsenosidases I type enzyme, Rb1, the Rb2 of conversion protopanoxadiol class, Rc and the preparation of Rd mixture.The protopanoxadiol saponins Rb1 of 100 grams, Rb2, Rc and Rd mixing saponin(e are dissolved in 1000 milliliters 0.02M, in the acetate buffer solution of pH5, with (by above-mentioned patent, the fermentation of literature method aspergillus tubigensis obtains) 1000 milliliters of mixing of ginsenosidases I type enzyme liquid, at 40 ℃ of stirring reaction 20-24 hours, add 2 liters macroporous resin (the resin HP-20 that anticipated, the Mitsubishi product), stirred gently one hour, saponin(e is adsorbed in the macroporous resin, wash the impurity such as zymoprotein and sugar off with deionized water, install in 3000 milliliters the post, wash impurity in the post with 10 premium on currency again, use again 12 liters 75% ethanol elution saponin(e; Its elutriant 1% decolorizing with activated carbon filters, and concentrating under reduced pressure, drying obtain content and be the F2 saponin(e crude products of 75% 80 grams; Obtaining content through the silicagel column separation again is the gram of 50 more than 90% F2 saponin(e, is used for the preparation of Rh2 group red ginseng saponin.
The GF2 of 25 grams is incorporated in 150 ml waters, grind, 100 milliliters of ginseng self enzymes and the extract of water-soluble substances and 300 milliliters the water that add again embodiment 1,80 ℃ of stirring reactions 8 hours, at 110 ℃ of enzymes 1 hour of going out, nearly all GF2 be converted into by red ginseng saponin 20 (
S)-Rh2,20 (
RThe Rh2 saponins that)-Rh2, Rk2 and Rh3 form.Its reaction solution through 0.5 liter macroporous resin (the HP-20 macroporous resin, MIT produces; Perhaps AB-8 macroporous resin, Tianjin Nankai university produces) post absorption saponin(e, with 4 liters sugar and the slag that does not adsorb that water-wash away; Use again 6 liters 80-84% ethanol elution saponin(e, its elutriant concentrating under reduced pressure, dry 17 grams 20 (
S)-Rh2,20 (
R)-Rh2, Rk2 and Rh3 mixing saponin(e, namely the Rh2 group is mixed saponin(e.
In the above-mentioned reaction, the reaction density 0.1-10% of GF2, the reaction ratio (weight) of the extract of GF2 and ginseng self enzyme and water-soluble substances (calculating of ginseng dry product raw material) is 1:0.1-20 times, temperature 70-120 ℃, all obtains good effect.
1, the affirmation of Rh2 group red ginseng saponin composition:
The high performance liquid chromatograph that Rh2 group red ginseng saponin is measured, U.S. waters 2695 instruments; Detector, Photodiode Array Detector Waters 2996; Knauer C18 chromatographic column (250mm * 3mm); Moving phase, acetonitrile (A)-water (B); 0 ~ 35min, the degree such as 19%A; 35 ~ 55min, 19%A ~ 29%A linear gradient; 55 ~ 65min, 29%A ~ 40%A linear gradient; 65 ~ 95min, 40%A ~ 100%A linear gradient; Sample size, 10 μ L; Column temperature, 35 ℃; Volumetric flow rate, 0.6mL/min; Detect wavelength, 203nm.
Get in the methyl alcohol that 2 milligrams of above-mentioned Rh2 group red ginseng saponins are dissolved in 1 milliliter, through 0.45 μ m membrane filtration, high performance liquid chromatography (HPLC) detects, with standard substance 20 (
S)-Rh2,20 (
R)-Rh2, Rk2 and Rh3 saponin(e compare, as shown in figure 13.
As can see from Figure 13, reaction obtain the Rh2 saponins contain 20 (
S)-Rh2,20 (
RFour kinds of rare saponin(es of red ginseng such as)-Rh2, Rk2 and Rh3, its content ratio (peak area ratio) is followed successively by 29:24:21:26.
2, Ultra Performance Liquid Chromatography-MS (UPLC-Q/TOF) is checked four kinds of saponin(es of Rh2 group:
Ultra Performance Liquid Chromatography-mass spectrometry analyser: UPLC-Q/TOF Premier
TMMicromass (Waters) instrument; Chromatographic column: BEH shield RP18 (2.1 * 100mm, 1.7 μ m, Waters).Ultra Performance Liquid Chromatography-quadrupole rods tandem mass spectrometry coupling detects and finishes at UPLC-Q/TOF PremierTM Micromass (Waters) instrument.The ESI source under the positive and negative ion pattern, is detected under " V " pattern.
Ultra Performance Liquid Chromatography (UPLC) testing conditions: moving phase is (A) 0.1% formic acid-acetonitrile---(B) 0.1% formic acid-water; Gradient elution: 0 ~ 5min, 5%A ~ 95%A linear gradient, 5 ~ 10min, the degree such as 95%A.Sample size, 10 μ L; Flow velocity, 0.5 mL/min; Column temperature, 30 ℃; Detect wavelength, 203nm.
Level Four time flight tandem mass spectrum (Q/TOF) testing conditions: capillary voltage, 2.5kv; Sample well voltage, 35v; Ion source temperature, 100 ℃; Desolventizing temperature degree, 350 ℃; The taper hole gas velocity, 50L/hr; The desolventizing gas velocity, 1000L/hr; The mass scanning scope, 100-1000Da.
The molecular formula of the rare saponin(e 20 of red ginseng (S)-Rh2 and 20 (R)-Rh2 is C
36H
62O
8, molecular weight is 622; The molecular formula C of Rk2 and Rh3
36H
60O
7, molecular weight is 604:
1) liquid-matter coupling method (UPLC-Q/TOF) detects the rare saponin(e of Rh2 group red ginseng as shown in figure 14.
As can be seen from Figure 14, Rh2 group detects in the collection of illustrative plates at UPLC four obvious ultraviolet absorption peaks, and Rh2 group HPLC 20 (
S)-Rh2,20 (
RIt is consistent that the ultraviolet absorption peak of)-Rh2, Rk2, Rh3 detects collection of illustrative plates, its retention time is followed successively by 4.62,4.66,5.33,5.38(min); Then under the negative ions pattern, detecting among the total ion current figure (province's sketch map) obtain by tandem mass spectrum (Q/TOF) has four obvious peaks, its retention time is followed successively by 4.62,4.71,5.36,5.41(min), consistent with the UPLC collection of illustrative plates, can carry out the total ion current peak and carry out the one-level mass spectroscopy, determine each material in the Rh2 group sample.
2) first, second quasi-molecular ions in the Rh2 saponins is carried out the one-level mass spectroscopy under the ESI-negative mode, shown in Figure 15,16.
20 (
S)-Rh2,20 (
R)-Rh2 molecular weight is 622; Can find out that from Figure 15,16 first, second peak of Rh2 saponins forms [first, second peak material molecule+HCOOH] under the ESI-negative mode
-Quasi-molecular ions, and reduce by a H under the negative ion mode, so the quasi-molecular ions m/z 667 that sample Rh2 obtains by Q/TOF.So can judge, contain 20 in the sample (
S)-Rh2 and 20 (
R)-Rh2, its molecular weight are 622.
Again the 3rd, the 4th quasi-molecular ions in the rare saponin(e of Rh2 group ginseng is carried out the one-level mass spectroscopy under the ESI-negative mode, shown in Figure 17,18.
The molecular weight of Rk2 and Rh3 is 604; From Figure 17,18, can find out, under the ESI-negative mode, form [the 3rd, the 4th peak material molecule+HCOOH]
-Quasi-molecular ions, and reduce by a H under the negative ion mode, so the quasi-molecular ions m/z 649 that Rk2 and Rh3 obtain.So can judge, contain Rk2 and Rh3 in the sample, its molecular weight is 604.Structural formula is respectively:
Proof again thus, contain 20 in the prepared Rh2 group (
S)-Rh2,20 (
RFour kinds of rare saponin(es of red ginseng such as)-Rh2, Rk2 and Rh2; Through several times test, in the prepared Rh2 group 20 (
S)-Rh2 and 20 (
RThe scope 20-80% of the content ratio of)-Rh2 in Rh2 group total saponins.
Embodiment 3:
Add organic acid in the extract of ginseng self enzyme and water-soluble substances, can improve and transform protopanoxadiol class Rb1, Rb2, Rc or Rd monomer or its mixing, preparation by 20 (
S)-Rg3,20 (
RThe effect that the Rg3 group that)-Rg3, Rk1 and Rg5 form is mixed the rare saponin(e of red ginseng.
5 gram ginsenoside Rb1s are incorporated in 22 ml waters, grind, add again ginseng self enzyme of 3 milliliters of embodiment 1 and the extract of water-soluble substances, add 25 ml waters and 10 gram citric acids, 70 ℃ of reactions 2 hours, dilute with 300 ml waters, on 100 milliliters macroporous resin column, repeatedly adsorb saponin(e, the water washery slag is used 400 milliliters 70% ethanol elution again, concentrating under reduced pressure, drying obtain 2.7 grams by 20 (
S)-Rg3,20 (
RThe Rg3 group that)-Rg3, Rk1 and Rg5 form is mixed saponin(e.
Several times evidence, in the above-mentioned reaction, the raw material ginsenoside can be used Rb1, Rb2, Rc, Rd monomer saponin, and also available its mixes saponin(e; Ginsenoside reaction density 0.1-10%, the reaction ratio (weight) of the extract of ginsenoside and ginseng self enzyme and water-soluble substances (calculating of ginseng dry product raw material) are 1:0.1-20 times; Used organic acid is lactic acid, acetic acid or citric acid, and the organic acid dosage is the 0.01-50% of reaction volume; Under the temperature of reaction 60-90 ℃ of condition, all obtain good effect.
Above-mentioned Rg3 group is mixed saponin(e and is detected through HPLC, its 20 (
S)-Rg3,20 (
R)-Rg3, Rk1 mix saponin(e with Rg5 saponin(e composition with the Rg3 group of embodiment 1 and form similar.
Embodiment 4:
Add organic acid in the extract of ginseng self enzyme and water-soluble substances, can improve conversion F2 and prepare Rh2 saponins effect.
5 gram GF2s were incorporated in 25 ml waters, grind, and add ginseng self enzyme of 5 milliliters of embodiment 1 and the extract of water-soluble substances again, add 15 milliliters of lactic acid, 80 ℃ of reactions 3 hours; Then with 300 ml waters dilutions, repeatedly adsorb saponin(e on 100 milliliters macroporous resin column, the water washery slag is used 400 milliliters 84% ethanol elution again, concentrating under reduced pressure, drying obtain 2.9 grams by 20 (
S)-Rh2,20 (
RThe Rh2 group that)-Rh2, Rk2 and Rh3 form is mixed saponin(e.
Several times evidence, in the above-mentioned reaction, GF2 reaction density 0.1-10%, the reaction ratio (weight) of the extract of GF2 and ginseng self enzyme and water-soluble substances (calculating of ginseng dry product raw material) they are 1:0.1-20 times; Used organic acid is lactic acid, acetic acid or citric acid, and the organic acid dosage is the 0.01-50% of reaction volume; Under the temperature of reaction 60-90 ℃ of condition, all obtain good effect.
Above-mentioned Rh2 group is mixed saponin(e and is detected through HPLC, its 20 (
S)-Rh2,20 (
R)-Rh2, Rk2 mix saponin(e with Rh3 red ginseng saponin composition with the Rh2 group of embodiment 2 and form similar.