CN112121071B - A product prepared from Ginseng radix - Google Patents
A product prepared from Ginseng radix Download PDFInfo
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- CN112121071B CN112121071B CN202011047127.0A CN202011047127A CN112121071B CN 112121071 B CN112121071 B CN 112121071B CN 202011047127 A CN202011047127 A CN 202011047127A CN 112121071 B CN112121071 B CN 112121071B
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Abstract
The invention relates to a product rich in rare ginsenoside, a preparation method and application. The product has high content of rare ginsenoside, and is substantially free of common ginsenoside. The preparation method is simple, does not need to use organic solvent and has no solvent residue. Also provides a rare ginsenoside composition with excellent anti-tumor effect.
Description
Technical Field
The invention relates to a product prepared by taking ginseng as a raw material, in particular to a ginseng product rich in rare ginsenoside.
Background
The ginseng is the dry root and rhizome of the ginseng of the panax of the Araliaceae, and the traditional commonly used rare Chinese medicinal materials in China are known as 'Baicaowang', have the advantages of mild nature, sweet taste, slight bitterness and slight temperature, have the efficacies of invigorating primordial qi, recovering pulse, relieving depletion, promoting the production of body fluid, quenching thirst, tonifying spleen, benefiting lung, calming the nerves, benefiting intelligence and the like, always occupy an extremely important position in the scientific research field and the traditional Chinese medicine market, and have considerable value in the aspects of medicine and health preservation.
The ginsenoside is a tetracyclic triterpene saponin extracted from plants (radix Panacis Quinquefolii, ginseng radix, notoginseng radix, etc.) of Panax of Araliaceae, rhizoma Panacis Majoris, rhizoma Panacis Japonici, and Vietnam radix. In the 60 s of the 20 th century, researches show that ginsenoside has anticancer activity, and the ginsenoside becomes a popular field for researching anticancer natural medicines. Later, the secondary metabolic derivatives of the transformed ginsenosides have stronger biological activity (Liurong: the research progress of preparing rare ginsenosides by the biotransformation technology). The secondary metabolic derivative is named as "rare ginsenoside", and is extracted directly from Panax plant of AraliaceaeThe ginsenoside is named as prototype ginsenoside, and the ginsenoside with high content is named as common ginsenoside, including ginsenoside Rg 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 Rd, etc. At present, more than 60 rare ginsenosides, including Rk, have been discovered 2 、Rg 3 、Rh 2 、Rg 5 、Rh 1 、Rh 3 、Rk 1 And various rare ginsenosides with different anticancer activities. Research shows that the rare ginsenoside has stronger antitumor activity, and also has pharmacological activities of reducing blood pressure, improving immunity, resisting inflammation and the like. Compared with common ginsenoside, the rare ginsenoside has better pharmacological activity.
In 2000, ginsenoside Rg has been approved in China 3 Ginsenoside Rh developed in 2006 for prescription medicine 2 The medicines are approved to be on the market, but the industrialization of the rare ginsenoside is still in the initial stage, and at present, no enterprise which can specially produce the high-purity rare ginsenoside monomer exists in China, and no enterprise which has the industrial production capacity of the rare ginsenoside exists. In the future, enterprises with the capacity of industrial production of rare ginsenoside must have the right of market speaking.
Since rare ginsenosides are secondary metabolites of ginsenosides, and the contents of them in panax plants are very low, and the artificial synthesis is difficult, so far, these rare ginsenosides can only be obtained by means of conversion of common ginsenosides, etc.
The conversion method of ginsenoside mainly comprises a chemical method, a microbiological method and an enzymatic method.
The chemical method comprises the following steps: ginsenoside-linked glycosyl groups can be cleaved when exposed to acids or bases. Under milder conditions, the glycosidic bond of ginsenoside is broken sequentially to obtain corresponding secondary ginsenosides, but if the conditions of hydrolysis reaction are too severe, for example, when strong acid such as sulfuric acid or hydrochloric acid is used as a reaction reagent, the sugar chain part of ginsenoside is completely hydrolyzed, and the glycosidic bond is completely broken to obtain ginsengenin, and even the structure of aglycon is changed. Dissolving ginsenoside Rd in Guo Yingying, etcHeating in formic acid solution with pH of 2.0 in water bath at 60 deg.C for 5h, separating and identifying ginsenoside Rd acid hydrolysate by liquid chromatography-quadrupole-time-of-flight mass spectrometry, and inferring chemical conversion mode of ginsenoside Rd by identifying the hydrolysate. The total saponins of caulis et folium Panacis Quinquefolii are subjected to alkaline hydrolysis with sodium hydroxide, and are subjected to ethyl acetate extraction, silica gel column chromatography and recrystallization to obtain ginsenoside Rh with high purity 1 And Rh 2 。
The microbiological method comprises the following steps: microbial conversion of ginsenosides refers to the process of converting a substrate into a target product through a special metabolic pathway by using microorganisms under appropriate conditions. In recent years, the improvement of the pharmacological activity of ginsenoside by microbial transformation has been receiving more and more attention. The microbial conversion method has the advantages of simple operation, mild conditions, high conversion efficiency of the ginsenoside, no need of purifying crude enzyme, no secondary pollution and the like. The glycosyl side chain of the ginsenoside is degraded by a microbiological method, so that various rare ginsenoside monomers can be obtained. Since 1987, the microbial transformation of 89 ginsenoside monomers of natural origin has been studied. The content of rare ginsenoside in radix Panacis Quinquefolii total saponin is increased by 20-150%, and various ginsenosides F are obtained by fermenting Chua Xiaoyu etc. with medicinal fungi 1 C-K and Rg 2 The content of rare ginsenoside is also obviously increased. Screening a strain of mold from Ming Yong shan et al, hydrolyzing ginsenoside Re to generate Rg 1 Separating and purifying with preparative column to obtain ginsenoside Rg with purity of 81.98% 1 The yield was 71.83%. Screening out TH-20 strain from soil such as coral, and simultaneously converting diol type ginsenoside and triol type ginsenoside to respectively obtain Rb 1 Re and Rg 1 Conversion to Rd, rg 2 And PPT. The strain CZ2 separated from orange juice such as Jinyan can convert ginsenoside Rb 1 Generating F 2 And Gyp-XVII. Zhangina et al use radix Panacis Quinquefolii extract as substrate to convert ginsenoside into ginsenoside, and analyze ginsenoside Re and its fermentation product by High Performance Liquid Chromatography (HPLC). The results show that the high-efficiency strainS329 can convert ginsenoside Re into ginsenoside Rh 1 The conversion was 27.65%. Cui Lei et al utilize endophyte JG09 to convert ginsenoside Rb 1 、Rb 2 、Rc、Rd、Rg 1 Respectively converted into corresponding rare ginsenoside F 2 C-K and Rh 1 Ginsenoside F 2 And the maximum conversion rate of C-K can reach 94.53 percent and 66.34 percent respectively.
An enzymatic conversion method: the enzyme method has the advantages of strong specificity, mild reaction conditions, high conversion efficiency, no pollution and the like, and is considered to be the most effective tool in the structural modification and metabolic research of the ginsenoside. The glycosidase is used for carrying out structural modification on the sugar chain to improve the biological activity of the compound so as to meet the development of the pharmaceutical industry, and is also the focus of the current ginsenoside research. Shin et al found that glucoside hydrolase in cellulolytic bacteria can hydrolyze D-glucose and L-arabinose outside the C-20 position of ginsenoside Rc, and thus ginsenoside Rc containing L-arabinose can be converted to C-K by this enzyme. The method comprises converting panaxadiol group ginsenoside with snailase to obtain rare ginsenoside C-K, and separating by silica gel column chromatography to obtain ginsenoside C-K with purity higher than 98%. Converting protopanaxatriol saponin with glycosidase to obtain rare ginsenoside F 1 And Rh 1 . Ginsenoside is used as a substrate and is subjected to enzymatic structure modification, so that the method is an effective way for preparing rare ginsenoside. Is prepared from ginsenoside Rb 2 、Rb 3 And Rc is used as a substrate, and C-3 site and C-20 site are modified to obtain rare ginsenoside Rg 3 、Rh 2 、F 2 And PPD and the like. The rare ginsenoside can be prepared in large scale, and the pharmacological activity of the ginsenoside is clear.
Chinese patent CN107468732A discloses a preparation method for increasing trace ginsenoside in traditional Chinese medicine ginseng leaf extract, which comprises the following steps: rubbing or crushing traditional Chinese medicine ginseng leaves by hands, mixing with diatomite =5, placing the mixture in an extraction kettle, placing the extraction kettle in a rapid solvent extraction instrument, taking an ethanol water solution as an extraction solution, wherein the volume ratio of the mixture of the ginseng leaves and the diatomite to ethanol is 30% -80%, the volume of the extraction solution is 60% -100% of the volume of the extraction kettle, the extraction temperature is 110 ℃ -200 ℃, extracting for 1-4 times, the extraction time is 1-20 minutes each time, combining the extraction solutions, shaking up, standing for 10-24 hours, taking supernate, concentrating and drying to obtain the ginseng leaf extract in a dry extract state. The method uses solvents such as diatomite, ethanol water and the like, and brings great troubles to subsequent purification and impurity removal.
The chemical method in the three ginsenoside conversion methods has the defects of difficult control of reaction conditions, complex product, environmental pollution and the like; the microbial transformation method has the defects of poor selectivity, difficult acquisition of microorganisms, incapability of applying some microorganisms to the food industry and the like; the enzyme method has the advantages of strong specificity, mild reaction conditions, high conversion efficiency, no pollution and the like, but the enzyme is very sensitive to temperature, often leads to inactivation, has higher requirements on the reaction conditions, and is difficult to realize industrial production.
Because of the difficulties in preparing rare ginsenosides, no ginseng product with high content of rare ginsenosides which can be produced in large scale has been available so far.
Therefore, there is a need in the art for a simple, easy to implement, low in pollution, low in organic solvent usage, low in residue, and purely natural ginseng product preparation method that can increase the content of rare ginsenoside and improve the bioactivity of ginseng products.
Disclosure of Invention
There are many methods for processing ginseng in the prior art, and accordingly, different ginseng products can be obtained.
A method comprises collecting Ginseng radix in autumn, cleaning, and sun drying or oven drying to obtain Ginseng radix divided into garden Ginseng radix and wild Ginseng radix; another method comprises collecting Ginseng radix in autumn, cleaning, steaming, and drying to obtain Ginseng radix Rubri. These two methods are the most traditional processing methods for ginseng. As a modern processing method of ginseng, a lactobacillus fermentation method, a high-temperature steaming method and the like appear, and products such as lactobacillus fermented ginseng or 'black ginseng' and the like can be obtained correspondingly. Another kind of ginseng products is various ginseng extracts obtained by extracting fresh ginseng or the ginseng products with water, ethanol or ethanol water, recovering the solvent from the extract under reduced pressure, and drying under vacuum. The other ginseng product is ginsenoside extract obtained by extracting the ginseng product with water, filtering, adsorbing the extract with macroporous adsorption resin, eluting with ethanol, recovering solvent from the eluate under reduced pressure, and vacuum drying.
CN108079033A extracting ginsenoside with 65-80% ethanol under reflux, filtering, discarding residue, and recovering solvent under reduced pressure to obtain total ginsenoside; CN106727806A is prepared by pulverizing folium Notoginseng, extracting with solvent, filtering, discarding residue, recovering solvent from the extractive solution under reduced pressure to obtain folium Notoginseng total saponin, wherein the solvent is selected from water and 70% ethanol. Weighing radix Panacis Quinquefolii coarse powder in CN102772462A, adding 70% ethanol solution 6 times the amount of the coarse powder each time, heating and reflux extracting for 3 times, filtering, discarding residue, and recovering solvent from the extractive solution under reduced pressure to obtain radix Panacis Quinquefolii total saponin.
Decocting with water or extracting under reflux with organic solvent, which is the most common method for extracting and separating effective components from Chinese medicinal materials or natural medicinal materials including ginseng to obtain Chinese medicinal material or natural medicinal extract. The specific operation process is that water is decocted and extracted (or organic solvent is refluxed and extracted), filtration is carried out, medicine dregs are discarded, extracting solution is decompressed and the solvent is recovered, and then vacuum drying is carried out to obtain the extract, which is the basic principle and the method for preparing the traditional Chinese medicine extract or the Chinese patent medicine. In order to obtain the best extraction conditions, factors such as an extraction solvent (water or an organic solvent), an extraction mode (soaking, heating reflux, percolation, ultrasound, supercritical), extraction times and the like are generally considered, so that the chemical components in the medicinal materials are completely extracted in the simplest and most economical mode. That is, in the research of chemical components of traditional Chinese medicine and the pharmaceutical engineering of traditional Chinese medicine, decocting the medicinal materials with water (or reflux extraction with organic solvent), centrifuging, discarding the extract, and taking the residue (dregs) after the decoction (or reflux extraction with organic solvent) as a new medicinal substance (product) is a matter contrary to the conventional wisdom, while finding that more chemical components which should enter the extract are retained in the residue (dregs) is not a matter which can be imagined by the conventional thinking.
The researchers of the invention have engaged in the research works such as the extraction of ginsenoside, the preparation of ginsenoside standard substance and effective part, the content measurement and the bioactivity, etc. for a long time. In the research process of using ginseng as raw material and water as solvent, reflux extracting ginsenoside in the ginseng and screening the optimal extraction process parameters, the influence of the decoction time on the transfer rate of the ginsenoside is repeatedly researched. During the initial extraction process, the content of the common ginsenoside in the water liquid (extract) is gradually increased along with the increase of the decoction time, and the content of the common ginsenoside in the dregs is gradually reduced, so that the result conforming to the general rule of extraction of chemical components of traditional Chinese medicines is obtained. However, in the following research process, it is unexpectedly found that, as the heating reflux extraction time is further prolonged, the content of the common ginsenoside in the water solution (extract) does not increase or decrease, and as the decoction time is further prolonged, the content of the common ginsenoside in the water solution (extract) continuously decreases, and after the water solution (extract) is heated for a certain time, especially 48 hours, the common ginsenoside is basically not contained in the water solution (extract); meanwhile, the rare ginsenoside begins to appear in the water solution and the decoction dregs and gradually rises along with the increase of the extraction time. It was then surprisingly found that after heating for a certain period of time, in particular 48 hours, the rare ginsenoside content in the aqueous liquid began to decrease and finally disappeared, while the rare ginsenoside content in the residue continued to increase and finally reached a maximum, and then the decoction time continued to be extended, the content did not substantially increase and did not decrease, with small fluctuations around the maximum. Based on the above important findings, through a great deal of elaborate researches, the invention finds a method for preparing a new ginseng product which contains more rare ginsenosides and basically does not contain common ginsenosides by taking ginseng as a raw material. Of course, this method can be used to obtain new ginseng products (dregs) containing different proportions of common ginsenosides and rare ginsenosides. The new ginseng product can be used for producing food, health food or medicinal product with health or therapeutic effect.
The ginseng product is preferably herb residue.
The rare ginsenoside comprises 20(S)-Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 、Rh 3 The common ginsenoside comprises Rg 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 And Rd. The product contains one or more of 15 kinds of rare ginsenoside, specifically 20 (S) -Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 、Rh 3 The product is substantially free of common ginsenoside including Rg 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 、Rd。
Preferably the total amount of rare ginsenosides is not less than 5%, 7%, 8%, 9%, 10%, 11% or 12%, and the total content of common ginsenosides is not more than 3%, 2%, 1%, 0.8%, 0.7%, 0.5%, 0.4%, 0.2% or 0.1%;
preferred among rare ginsenosides Rg 3 The content is not less than 3%, more preferably not less than 4% or 6%, most preferably not less than 6%.
Preferred rare ginsenosides are Rk 1 And Rg 5 The sum of the amounts is not less than 1%, more preferably not less than 2%, 3%, 4%, or 5%.
Preferred rare ginsenosides are Rk 6 And F 4 The sum of the contents is not less than 0.5%, more preferably not less than 0.6% or 0.7%.
Preferably the total content of common ginsenosides is not higher than 1%, more preferably not higher than 0.8% or 0.6%, most preferably not higher than 0.4% and 0.3%.
Preferred common ginsenoside Rc contents are not higher than 3%, 2%, 1%, 0.8%, 0.7%, 0.5%, 0.4%, 0.2% or 0.1%.
Preferred common ginsengSaponin Rg 1 、Re、Rb 1 、Rb 2 、Rb 3 Rd, more preferably not more than 0.4% or 0.3%, most preferably not more than 0.2%, 0.1% or 0.05%.
Rg described in the present invention 3 、Rh 1 Or Rh 2 Including the 20R configuration and/or the 20S configuration, the ratio of the 20R configuration to the 20S configuration is 1.
Rg described in the present invention 3 Comprises 20 (S) -Rg 3 And/or 20 (R) -Rg 3 ；Rh 2 Comprising 20 (S) -Rh 2 And/or 20 (R) -Rh 2 。
The application also relates to a preparation method for preparing the high-content rare ginsenoside product, which comprises any one of the following steps:
a: pulverizing Ginseng radix, adding appropriate amount of water, heating and refluxing under normal pressure for more than 48 hr, preferably 60 hr, 72 hr, 84 hr, 96 hr 108 hr or more than 120 hr, filtering, and drying the residue to obtain the desired product;
b: pulverizing Ginseng radix, adding appropriate amount of water, decocting at high temperature and high pressure for 4 hr or more, preferably 6 hr or more, 8 hr or more, 10 hr or more, 12 hr or more, 14 hr or more, 16 hr or more, 18 hr or more, 20 hr or more, 22 hr or more, 24 hr or more, 28 hr or more, 30 hr or more, 32 hr or more, 34 hr or more, 36 hr or more, 38 hr or more, 40 hr or more, 42 hr or more, 44 hr or more, 48 hr or more, or 50 hr or more, filtering, and drying the residue to obtain the desired product.
The high temperature is 100 deg.C or higher, preferably not lower than 110 deg.C, 115 deg.C, 120 deg.C, 125 deg.C, 130 deg.C, 135 deg.C, 140 deg.C, 145 deg.C or 150 deg.C.
In preparation method A, the liquid-to-material ratio can be (30-5): 1, preferably (25-8): 1, more preferably (20-9): 1 or (15-10): 1, preferably 10.
In the preparation method B, the liquid-to-material ratio can be (30-5): 1, preferably (25-6): 1, more preferably (20-7): 1 or (10-8): 1, preferably 7.5).
In the preparation method A, preferably, the ginseng is first soaked in water; more preferably, ginseng is soaked for 6 hours or more, 8 hours or more, 10 hours or more, 12 hours or more, or 24 hours or more, and then heated under reflux; while filtering while hot after the completion of the decoction, it is most preferable to filter while hot (near 100 ℃ C.) immediately after the completion of the decoction.
In the preparation method B, the water is preferably heated and then fed, the water is preferably heated to above 60 ℃ and then fed, and the water is more preferably heated to above 90 ℃ and then fed; most preferred is to heat the water to 100 ℃ before adding it. And filtering while the decoction is hot, and most preferably filtering while the decoction is hot (nearly 100 ℃) immediately after the decoction.
20 (S) -Rg in the medicine dregs 2 The content range of (A) is 0.2% -0.35%;20 (S) -Rh 1 The content range is 0-0.45%, preferably 0.2% -0.45%;20 (R) -Rh 1 The content range is 0-0.25%, preferably 0.05% -0.25%; rg (Rg) 6 The content range is 0-0.15%, preferably 0.1% -0.15%; f 4 The content range is 0-0.8%, preferably 0.1% -0.7%; rk 3 The content range is 0-0.2%, preferably 0.02% -0.2%; rh 4 The content range is 0-0.1%, preferably 0.05% -0.35%;20 (S) -Rg 3 The content range is 0.2% -4.0%, preferably 0.4% -3.8%;20 (R) -Rg 3 The content range is 0.2% -3.0%, preferably 0.3% -2.8%; rk 1 The content range is 0.1% -2.0%, preferably 0.2% -2.0%; rg (Rg) 5 The content range is 0.2% -3.0%, preferably 0.3% -2.8%;20 (S) -Rh 2 The content range is 0-0.07%;20 (R) -Rh 2 The content range is 0-0.05%; rk 2 The content range is 0-0.04%; rh 3 The content range is 0-0.06%.
Optionally further preparing the residue into powder, decoction pieces, syrup, oral liquid, suspension, emulsion, tablet, capsule, delayed release agent, quick release agent, controlled release agent, injection, injectable powder, patch, suppository, or drop.
Optionally further extracting the residue with organic solvent or aqueous organic solvent to obtain extract with high content of rare ginsenoside.
The preferred extract contains one or more of 15 rare ginsenosides, specifically 20 (S) -Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 、Rh 3 The total content of rare ginsenosides is more than 15%, 20%, 25%, 30%, 35%, 40%, 50%, 60%, 70%, 80% or 90%. Preferably, the extract contains Rg 3 (20S + 20R) content is not less than 15%, not less than 18%, not less than 20% or not less than 22%; and/or Rg 5 The content of (A) is not less than 5%, 6%, 8%, or 9%; and/or Rk 1 The content of (A) is not less than 5% or 6%; and/or F 4 The content of (A) is not less than 1%; rg (Rg) 6 And F 4 The sum of the contents of (A) and (B) is not less than 1.5% or 2%.
More preferably, the extract is substantially free of common ginsenosides, including Rg 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 And Rd. The content of common ginsenoside is not higher than 10%, preferably not higher than 8%, 6%, 5%, 4%, 3%, 2% or 1%. More preferably, less than 0.8%, 0.6%, 0.5% or 0.1%.
20 (S) -Rg in the extract 2 The content range of (A) is 0.2% -1.5%;20 (S) -Rh 1 The content range is 0.5% -1.6%;20 (R) -Rh 1 The content range is 0.5% -0.8%; rg (Rg) 6 The content range is 0.3% -0.8%; f 4 The content range is 1% -2.5%; rk 3 The content range is 0.5% -1.2%; rh 4 The content range is 0.5% -1.6%;20 (S) -Rg 3 The content range is 5% -13%;20 (R) -Rg 3 The content range is 5% -12%; rk 1 The content range is 5% -10%; rg (Rg) 5 The content range is 6% -12%;20 (S) -Rh 2 The content range is 0.1% -0.3%;20 (R) -Rh 2 The content range is 0.1% -0.3%; rk 2 The content range is 0.05％-0.2％；Rh 3 The content range is 0.1% -0.4%.
The organic solvent may be methanol, ethanol, ethyl acetate, chloroform, n-butanol, acetone or isopropanol.
The present application also provides a composition comprising rare ginsenosides, preferably the composition has synergistic anti-tumor activity.
The rare ginsenoside comprises Rg 3 And Rg 5 The composition of (1) to (20): (1-20), preferably (2-20): (2-20); (4-6) and (1-3); or (7-11.5) to (3.5-5).
The rare ginsenoside comprises F 4 And Rh 2 The composition of (1) to (100): (1-100), preferably (1-10): (10-100), (10-100): (1-10), (1-10): (10-1), (1-100): (1-10); most preferred is 10.
The rare ginsenoside contains Rh 4 And Rh 2 The composition of (1) to (100): (1-100), preferably (1-90): (1-90), (10-100): (1-10), (1-10): (10-1), (1-20): (1-20).
The rare ginsenoside in the composition optionally further comprises S-Rg 2 、S-Rh 1 、R-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、S-Rg 3 、R-Rg 3 、Rk 1 、Rg 5 、S-Rh 2 、R-Rh 2 、Rk 2 、Rh 3 One or more rare ginsenosides in (1).
Preferably, the composition also comprises 20 (S) -Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 And/or Rh 3 。
Preferably, the composition also comprises 20 (S) -Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 、Rh 3 。
Preferably, the composition is substantially free of common ginsenosides, including Rg 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 And Rd. The content of common ginsenoside is less than 10%, preferably less than 8%, 6%, 5%, 4%, 3%, 2% or 1%. More preferably, less than 0.8%, 0.6%, 0.5% or 0.1%.
The application also provides a ginseng product containing any one of the compositions, preferably a product prepared by taking natural ginseng or common ginsenoside as raw materials, preferably herb residue or extract.
Preferably, the properties and parameters of the product are characterized as any one of the above.
The ginseng comprises one or more of dried or fresh ginseng root, ginseng rootlets and ginseng reed head.
The application also provides the application of the product or the composition in treating and/or preventing tumors, wherein the tumors can be lung cancer, colon cancer and glioma.
The application of any one of the above products or compositions in the preparation of medicines, health foods or foods, in particular to the application in foods, health foods or medicines which are known in the prior art and used for reducing blood sugar, resisting fatigue and viruses, and treating diabetes, cardiovascular diseases, central nervous diseases, biological metabolism, reproductive function, immune function, renal function and other diseases; which may contain pharmaceutically or food acceptable carriers or additives.
Any of the products or compositions described above of the present application, which may contain a pharmaceutically acceptable carrier.
Optionally, the product or composition is administered orally, by injection or transdermally.
Optionally, the product or composition is in the form of powder, decoction pieces, syrup, oral liquid, suspension, emulsion, tablet, capsule, sustained release preparation, quick release preparation, controlled release preparation, injection, injectable powder, patch, suppository, or drop.
Drawings
FIG. 1: the ginseng rootlets dregs react at 100 ℃ and Rg in water 3 The content is changed;
FIG. 2: the ginseng rootlets dregs react at 100 ℃ and Rg in water 3 And Rh 2 A change in total amount;
FIG. 3: reacting ginseng rootlets dregs and Rk in water at 100 DEG C 1 And Rg 5 A change in total amount;
FIG. 4: reacting Leptoradix Ginseng residue at 100 deg.C and Rg in water 6 And F 4 A change in total amount;
FIG. 5: reacting ginseng rootlets dregs at 100 ℃ to neutralize the total amount change of 15 rare ginsenosides in water;
FIG. 6: reacting ginseng rootlets dregs and 7 common ginsenosides in water at 100 ℃;
FIGS. 7A and 7B show the total amount of 15 rare ginsenosides in the residue at 115 deg.C and 120 deg.C, respectively;
fig. 8A and 8B are respectively: the total amount of 15 rare ginsenosides in the residue at 125 deg.C and 130 deg.C is changed;
fig. 9A and 9B are respectively: the total amount of 15 rare ginsenosides in the residue at 135 deg.C and 140 deg.C is changed;
FIG. 10 is a diagram: the total amount of 15 rare ginsenosides in the residue is changed at 145 ℃;
FIG. 11: 20 (S) Rg in medicine dregs at 115 DEG C 3 +20(R)Rg 3 And Rg 5 +Rk 1 The content is changed;
FIG. 12: 20 (S) Rg in the medicine dregs at 120 DEG C 3 +20(R)Rg 3 And Rg 5 +Rk 1 The content is changed;
FIG. 13: 20 (S) Rg in medicine dregs at 125 DEG C 3 +20(R)Rg 3 And Rg 5 +Rk 1 The content is changed;
FIG. 14: 20 (S) Rg in dregs of a decoction at 130 DEG C 3 +20(R)Rg 3 And Rg 5 +Rk 1 The content is changed;
FIG. 15: 20 (S) Rg in dregs of a decoction at 135 DEG C 3 +20(R)Rg 3 And Rg 5 +Rk 1 The content is changed;
FIG. 16: 20 (S) Rg in dregs of a decoction at 140 DEG C 3 +20(R)Rg 3 And Rg 5 +Rk 1 The content is changed;
FIG. 17: 20 (S) Rg in medicine dregs at 145 DEG C 3 +20(R)Rg 3 And Rg 5 +Rk 1 The content was varied.
Detailed Description
The present invention will be further described with reference to the following examples.
Determining the content of common ginsenoside by high performance liquid chromatography:
1. ginsenoside Rg 1 、Re、Rb 2 、Rb 3 Determination of the content of Rd
1.1 chromatographic conditions
Octadecylsilane chemically bonded silica gel as filler (column length 250mm, inner diameter 4.6mm, particle size 5 μm), acetonitrile as mobile phase A, water as mobile phase B, gradient elution according to the following table, column temperature 25 deg.C,
the flow rate is 1ml/min, the detection wavelength is 203nm, and the sample injection amount is 20 mu L.
| Time (minutes) | Mobile phase A (%) | Mobile phase B (%) |
| 0～35 | 19.5 | 80.5 |
| 35～35.1 | 19.5→20 | 80.5→80 |
| 35.1～55 | 20 | 80 |
| 55～55.1 | 20→29 | 80→71 |
| 55.1～84 | 29→36 | 27→64 |
| 84～90 | 36 | 64 |
1.2 preparation of control solutions:
collecting ginsenoside Rg 1 Ginsenoside Re and ginsenoside Rb 2 Ginsenoside Rb 3 And a proper amount of ginsenoside Rd reference substance, precisely weighing, and adding methanol to obtain a mixed standard solution with the following concentration.
Ginsenoside Rg 1 The concentration is 0.201mg/ml
The ginsenoside Re concentration is 0.205mg/ml
Ginsenoside Rb 2 The concentration is 0.174mg/ml
Ginsenoside Rb 3 The concentration is 0.111mg/ml
The concentration of ginsenoside Rd is 0.201mg/ml
1.3 preparation of test solution:
1.3.1, herb residue
Taking 2.0g of sample (dried ginseng dregs), precisely weighing, adding methanol for reflux extraction twice, wherein the amount of the methanol is 250ml, each time lasts for 2 hours, filtering, recovering the methanol from the filtrate under reduced pressure, and metering the volume of the residue to 100ml by using the methanol.
1.3.2, water solution
Taking 25ml of water solution, adding methanol to a volume of 50ml volumetric flask.
2. Ginsenoside Rb 1 Measurement of Rc content
2.1 chromatographic conditions:
a chromatographic column: XAmide column (250 mm column length, 4.6mm inner diameter, 5 μm particle size), acetonitrile as mobile phase A, water as mobile phase B, according to the following table. The column temperature was 25 ℃, the flow rate was 1ml/min, the detection wavelength was 203nm, and the sample volume was 20. Mu.L.
| Time (minutes) | Mobile phase A (%) | Mobile phase B (%) |
| 0～30 | 90→84 | 10→16 |
| 30～65 | 84 | 16 |
2.2 preparation of control solutions:
collecting ginsenoside Rb 1 And a proper amount of ginsenoside Rc reference substance is precisely weighed and added with methanol to prepare a mixed standard solution with the following concentration. Ginsenoside Rb 1 The concentration is 0.121mg/ml
Ginsenoside Rc concentration is 0.156mg/ml
2.3 preparation of test solution:
2.3.1, herb residue
Taking 2.0g of sample (dried ginseng residue), precisely weighing, extracting with methanol under reflux twice each for 2 hr, filtering, discarding residue, recovering methanol from filtrate under reduced pressure, and diluting residue with methanol to 100ml.
2.3.2, water solution
Taking 25ml of water solution, adding methanol to a volume of 50ml volumetric flask.
High performance liquid chromatography for determining rare ginsenoside content
1. Chromatographic conditions are as follows:
octadecylsilane chemically bonded silica (ODS) was used as a filler (column length 250mm, inner diameter 4.6mm, particle size 5 μm), acetonitrile was used as a mobile phase A, water was used as a mobile phase B, and gradient elution was performed according to the following table, with a column temperature of 25 ℃, a flow rate of 0.8ml/min, a detection wavelength of 203nm, and a sample introduction amount of 20 μ L.
| Time (minutes) | Mobile phase A (%) | Mobile phase B (%) |
| 0～10 | 29 | 71 |
| 10～25 | 29→40 | 71→60 |
| 25～50 | 40→60 | 60→40 |
| 50～60 | 60→73 | 40→27 |
| 60～75 | 73 | 27 |
2. Preparation of control solutions:
collecting ginsenoside 20 (S) -Rg 2 Ginsenoside 20 (S) -Rh 1 And (20) ginsenoside R-Rh 1 Ginsenoside Rg 6 Ginsenoside F 4 Ginsenoside Rk 3 Ginsenoside Rh, ginsenoside Rh 4 Ginsenoside 20 (S) -Rg 3 Ginsenoside 20 (R) -Rg 3 Ginsenoside Rk 1 Ginsenoside Rg 5 Ginsenoside 20 (S) -Rh 2 Ginsenoside 20 (R) -Rh 2 Ginsenoside Rk 2 Ginsenoside Rh, ginsenoside Rh 3 The appropriate amount of reference substance is precisely weighed and added with methanol to prepare mixed standard solution with the following concentration.
Ginsenoside 20 (S) -Rg 2 The concentration is 0.2268mg/ml
Ginsenoside 20 (S) -Rh 1 The concentration is 0.1792mg/ml
Ginsenoside 20 (R) -Rh 1 The concentration is 0.1692mg/ml
Ginsenoside Rg 6 The concentration is 0.1632mg/ml
The concentration of ginsenoside F4 is 0.1240mg/ml
Ginsenoside Rk 3 The concentration is 0.1248mg/ml
Ginsenoside Rh 4 The concentration is 0.1256mg/ml
Ginsenoside 20 (S) -Rg 3 The concentration is 0.1296mg/ml
Ginsenoside 20 (R) -Rg 3 The concentration is 0.0596mg/ml
Ginsenoside Rk 1 The concentration is 0.1580mg/ml
Ginsenoside Rg 5 The concentration is 0.1560mg/ml
Ginsenoside 20 (S) -Rh 2 The concentration is 0.1388mg/ml
Ginsenoside 20 (R) -Rh 2 The concentration is 0.1324mg/ml
Ginsenoside Rk 2 The concentration is 0.1324mg/ml
Ginsenoside Rh 3 The concentration is 0.0688mg/ml
3. Preparing a test solution:
3.1, herb residue
Taking 3.0g of dried ginseng dregs, precisely weighing, carrying out reflux extraction twice with methanol for 2 hours each time, wherein the amount of the methanol is 250ml, filtering, discarding the dregs, recycling the methanol from the filtrate under reduced pressure, and using the methanol to fix the volume of the residue to a volumetric flask of 250 ml.
3.2, water solution
Taking 25ml of water solution, adding methanol to a volume of 50ml volumetric flask.
Example 1: decocting Ginseng radix at 100 deg.C under normal pressure-distribution of ginsenoside in water solution and residue
Weighing 1500g of ginseng rootlets as raw materials, crushing, adding 10 times of water for soaking overnight, heating and refluxing at 100 ℃ (normal pressure), and sampling and filtering respectively to obtain a water solution (liquid) and dregs, wherein the water solution (liquid) and dregs are dried to obtain dry dregs after 0 hour (before the soaking overnight begins to heat and reflux), 60min after the beginning of heating, A (90 min, just begins to boil), 1 hour, 2 hours, 3 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 24 hours, 36 hours, 48 hours, 60 hours, 72 hours, 84 hours, 96 hours, 108 hours, 120 hours, 132 hours, 144 hours, 156 hours, 168 hours, 180 hours, 192 hours, 204 hours, 216 hours, 228 hours, 240 hours and 252 hours. And analyzing the contents of the rare ginsenoside and the common ginsenoside in the water solution and the medicine residue by using 15 rare ginsenosides and 7 common ginsenosides as standard substances through an HPLC method. 600g ginseng rootlets are heated and refluxed for 84 hours to obtain 201g of dry medicine dregs, and 5750ml of centrifugal water is obtained.
I. Common ginsenoside content change in water
TABLE 1 content change of ginsenoside in 252 hr water solution of ginseng rootlets refluxed at 100 deg.C under normal pressure (mg/ml, content is 0 after 96 hr)
TABLE 2 content of ginsenoside in ginseng rootlets after refluxing for 252 hours at 100 deg.C under normal pressure (%, the content is 0 after refluxing for 132 hours)
From the above table, it can be seen that, after ginseng is soaked in 10 times of water for 12 hours, the ginseng begins to be heated and decocted, and is heated to be within 4.5 hours (boiling begins for 3.0 hours), the content of the common ginsenoside in the water solution keeps rising, the content of the common ginsenoside in the water solution reaches the maximum (4.399 mg/ml) after boiling for 3 hours, then the content of the common ginsenoside in the water solution begins to fall, only one common ginsenoside is detected in 84 hours, the content is only 0.05%, and no common ginsenoside is detected in 96 hours. Correspondingly, the content of the common ginsenoside in the medicine residue is gradually reduced, only 2 common ginsenosides are detected in 120 hours, only 1 common ginsenosides is detected in 132 hours, the content of the common ginsenosides can be basically ignored, and the content of the common ginsenosides in 144 hours is detected to be 0.
TABLE 3 content (mg/ml) of 15 rare ginsenosides in ginseng rootlets reflux water solution at 100 deg.C under normal pressure for 252 hours
TABLE 4 content (%) of 15 rare ginsenosides in the residue after refluxing Leptoradix Ginseng at 100 deg.C under normal pressure for 252 hr
Rare ginsenoside can not be detected in the water liquid and the dregs of a decoction of the ginseng rootlets after being soaked for one night, which indicates that the ginseng rootlets originally do not contain rare ginsenoside.
When the water solution is heated to boiling, rare ginsenoside begins to appear in the water solution, and the content of rare ginsenoside in the water solution gradually increases with the increase of the heating time until the content reaches the maximum (2.897 mg/ml) after 36 hours. Then the content of rare ginsenoside in water solution begins to gradually decrease to 0.127mg/ml after 252h, and decreases to less than 5% of the peak.
Correspondingly, rare ginsenoside begins to appear in the medicine residue when the medicine residue is heated to be boiled, the content of the rare ginsenoside gradually increases to 12.431% when the medicine residue is heated to be boiled for 84h, and then the content of the rare ginsenoside shows a small fluctuation trend around the value.
Rare ginsenoside Rg 3 (20(S)Rg 3 +20(R)Rg 3 ) And Rh 2 (20(S)Rh 2 +20(R)Rh 2 ) Before refluxing for 36 hours, the content of the water solution is increased and gradually decreased after 36 hours, and after 120 hours, the content is negligibly low.
Rg in herb residues 3 And Rh 2 The content of (A) gradually rises, and the content can reach more than 5 percent; rg (Rg) 3 And Rh 2 The total content of the two components can reach more than 5 percent, and particularly after 84 hours, the sum of the two components can reach 6.3 percent at most.
In addition, rk in rare ginsenoside 1 And Rg 5 The total amount can reach more than 4 percent. Rare personRg in ginsenoside 6 And F 4 The content sum is not less than 0.5 percent and can reach more than 0.6 percent or 0.7 percent.
In order to maximally enrich the rare ginsenoside in the medicine residues, reduce the loss of the rare ginsenoside in water liquid and shorten the preparation time, the ginseng fibrous root decoction takes ginseng rootlets as raw materials and carries out a high-temperature high-pressure decoction experiment.
Example 2: high-temperature high-pressure decoction of Leptoradix Ginseng, distribution of ginsenoside in water solution and residue
Taking Leptoradix Ginseng as raw material, weighing 1000 g Leptoradix Ginseng, pulverizing, adding 7.5 times of water, decocting at high temperature and high pressure, sampling at a certain time interval from 0 hr (the temperature in the kettle reaches a set temperature), filtering to obtain residue and water solution, and oven drying the residue to obtain dry residue. Analyzing the contents of rare ginsenoside and common ginsenoside in water solution and residue by HPLC.
The temperature is respectively designed to be 115 ℃,120 ℃, 125 ℃, 130 ℃, 135 ℃,140 ℃ and 145 ℃, the liquid-material ratio is designed to be 5.
Compared with the high-temperature high-pressure decoction and the normal-pressure reflux at 100 ℃, the change rule of common ginsenoside and rare ginsenoside in the water solution and the medicine residue is basically consistent, and the content of the rare ginsenoside in the prepared medicine residue is basically the same, but the high-temperature high-pressure decoction has the advantage of greatly shortening the preparation time compared with the normal-pressure reflux.
Experiments show that Rg in medicine dregs under the conditions of high temperature and high pressure 3 The content of Rg in the medicine residue reaches more than 5 percent, especially when the temperature is 125-140 ℃, the liquid-material ratio is (6-9): 1 3 The content of (A) can reach more than 6%. After a period of decoction, the contents of common ginsenoside in the dregs of a decoction and water are both 0.
The following were performed for experimental data at 115 ℃,120 ℃, 125 ℃, 130 ℃, 135 ℃,140 ℃ and 145 ℃ (all liquid-to-material ratios are 7.5).
Table 5: the contents of 7 common ginsenosides in ginseng rootlet dregs decocted at the high temperature and the high pressure of 115%
| 2h | 4h | 6h | 8h | 12h | 16h | 20h | 24h | 28h | 32h | 36h | 40h | 44h | 48h | 52h | |
| Rg 1 | 0.061 | 0.042 | 0.028 | 0.015 | 0.003 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Re | 0.273 | 0.181 | 0.112 | 0.058 | 0.017 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rb 1 | 1.168 | 0.972 | 0.915 | 0.738 | 0.470 | 0.170 | 0.085 | 0.049 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rc | 0.955 | 0.802 | 0.734 | 0.588 | 0.346 | 0.147 | 0.065 | 0.027 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rb 2 | 0.618 | 0.511 | 0.477 | 0.397 | 0.290 | 0.097 | 0.051 | 0.031 | 0.010 | 0.004 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rb 3 | 0.093 | 0.076 | 0.070 | 0.057 | 0.043 | 0.014 | 0.007 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rd | 0.547 | 0.498 | 0.492 | 0.448 | 0.402 | 0.212 | 0.174 | 0.161 | 0.105 | 0.068 | 0.036 | 0.027 | 0.000 | 0.000 | 0.000 |
| Total amount (%) | 3.715 | 3.082 | 2.828 | 2.301 | 1.571 | 0.640 | 0.382 | 0.268 | 0.115 | 0.072 | 0.036 | 0.027 | 0.000 | 0.000 | 0.000 |
Table 6: decocting Leptoradix Ginseng at 115 deg.C under high temperature and high pressure to obtain 7 common ginsenoside contents (mg/ml, 4 hr, 8 hr and 48 hr for three time points)
| 4h | 8h | 48h | |
| Rg 1 | 0.138 | 0.048 | 0.000 |
| Re | 0.508 | 0.175 | 0.000 |
| Rb 1 | 1.543 | 1.031 | 0.000 |
| Rc | 1.276 | 0.807 | 0.000 |
| Rb 2 | 0.829 | 0.540 | 0.000 |
| Rb 3 | 0.122 | 0.084 | 0.000 |
| Rd | 0.694 | 0.530 | 0.000 |
| Total amount (mg/ml) | 5.110 | 3.215 | 0.000 |
TABLE 7 15 rare ginsenoside contents (%) -in Leptoradix Ginseng residue decocted at 115 deg.C under high temperature and high pressure
| 2h | 4h | 6h | 8h | 12h | 16h | 20h | 24h | 28h | 32h | 36h | 40h | 44h | 48h | 52h | |
| S-Rg 2 | 0.183 | 0.233 | 0.282 | 0.307 | 0.338 | 0.281 | 0.298 | 0.314 | 0.320 | 0.324 | 0.298 | 0.357 | 0.219 | 0.235 | 0.169 |
| S-Rh 1 | 0.405 | 0.407 | 0.432 | 0.421 | 0.404 | 0.279 | 0.280 | 0.304 | 0.321 | 0.345 | 0.342 | 0.445 | 0.316 | 0.360 | 0.288 |
| R-Rh 1 | 0.025 | 0.034 | 0.038 | 0.048 | 0.068 | 0.063 | 0.081 | 0.096 | 0.112 | 0.131 | 0.140 | 0.203 | 0.184 | 0.225 | 0.199 |
| Rg 6 | 0.029 | 0.047 | 0.063 | 0.074 | 0.091 | 0.074 | 0.089 | 0.105 | 0.123 | 0.138 | 0.132 | 0.166 | 0.124 | 0.136 | 0.118 |
| F 4 | 0.063 | 0.099 | 0.134 | 0.157 | 0.204 | 0.175 | 0.213 | 0.262 | 0.324 | 0.373 | 0.393 | 0.561 | 0.417 | 0.514 | 0.428 |
| Rk 3 | 0.016 | 0.022 | 0.029 | 0.034 | 0.042 | 0.040 | 0.050 | 0.062 | 0.079 | 0.095 | 0.105 | 0.159 | 0.149 | 0.179 | 0.158 |
| Rh 4 | 0.025 | 0.033 | 0.044 | 0.050 | 0.067 | 0.063 | 0.079 | 0.096 | 0.127 | 0.155 | 0.172 | 0.268 | 0.235 | 0.308 | 0.279 |
| S-Rg 3 | 0.196 | 0.328 | 0.482 | 0.637 | 1.041 | 0.990 | 1.335 | 1.724 | 2.077 | 2.408 | 2.550 | 3.560 | 2.781 | 3.348 | 2.730 |
| R-Rg 3 | 0.116 | 0.186 | 0.300 | 0.452 | 0.926 | 0.605 | 0.946 | 1.426 | 1.057 | 1.460 | 1.711 | 2.324 | 2.419 | 2.713 | 1.695 |
| Rk 1 | 0.096 | 0.162 | 0.238 | 0.315 | 0.523 | 0.470 | 0.639 | 0.864 | 1.081 | 1.254 | 1.345 | 1.912 | 1.487 | 1.839 | 1.535 |
| Rg 5 | 0.141 | 0.231 | 0.337 | 0.444 | 0.734 | 0.658 | 0.899 | 1.233 | 1.551 | 1.805 | 1.953 | 2.784 | 2.167 | 2.712 | 2.296 |
| S-Rh 2 | 0.020 | 0.021 | 0.022 | 0.024 | 0.026 | 0.025 | 0.031 | 0.037 | 0.045 | 0.052 | 0.058 | 0.076 | 0.051 | 0.071 | 0.068 |
| R-Rh 2 | 0.012 | 0.014 | 0.013 | 0.015 | 0.019 | 0.020 | 0.025 | 0.028 | 0.028 | 0.033 | 0.031 | 0.048 | 0.030 | 0.032 | 0.025 |
| Rk 2 | 0.000 | 0.000 | 0.000 | 0.000 | 0.002 | 0.001 | 0.003 | 0.006 | 0.009 | 0.010 | 0.013 | 0.029 | 0.029 | 0.038 | 0.024 |
| Rh 3 | 0.000 | 0.000 | 0.004 | 0.005 | 0.007 | 0.008 | 0.011 | 0.015 | 0.022 | 0.026 | 0.029 | 0.048 | 0.027 | 0.048 | 0.052 |
| Rg 3 Total amount of | 0.313 | 0.513 | 0.783 | 1.089 | 1.967 | 1.595 | 2.281 | 3.150 | 3.134 | 3.867 | 4.260 | 5.884 | 5.200 | 6.061 | 4.425 |
| Rh 2 Total amount of | 0.032 | 0.034 | 0.036 | 0.038 | 0.046 | 0.045 | 0.056 | 0.065 | 0.073 | 0.085 | 0.090 | 0.124 | 0.081 | 0.103 | 0.093 |
| Rg 3 +Rh 2 | 0.345 | 0.548 | 0.818 | 1.127 | 2.012 | 1.640 | 2.337 | 3.216 | 3.207 | 3.952 | 4.350 | 6.008 | 5.280 | 6.163 | 4.518 |
| Rk 1 +Rg 5 | 0.237 | 0.393 | 0.575 | 0.759 | 1.257 | 1.128 | 1.538 | 2.097 | 2.632 | 3.059 | 3.298 | 4.696 | 3.654 | 4.551 | 3.831 |
| Rg 6 +F 4 | 0.092 | 0.146 | 0.197 | 0.231 | 0.295 | 0.249 | 0.302 | 0.367 | 0.447 | 0.511 | 0.525 | 0.727 | 0.541 | 0.650 | 0.546 |
| Total (%) | 1.327 | 1.817 | 2.418 | 2.983 | 4.492 | 3.752 | 4.979 | 6.572 | 7.276 | 8.609 | 9.272 | 12.940 | 10.635 | 12.758 | 10.064 |
Table 8: decocting Leptoradix Ginseng at 115 deg.C under high temperature and high pressure to obtain 15 kinds of rare ginsenoside contents (mg/ml, 4 hr, 8 hr and 48 hr for three time points)
Table 9: 15 rare ginsenoside contents in ginseng rootlets dregs decocted at high temperature and high pressure of 125 (%)
| 1h | 6h | 8h | 12h | 16h | 20h | 24h | 28h | 32h | 36h | 40h | 44h | 48h | |
| S-Rg 2 | 0.209 | 0.288 | 0.284 | 0.359 | 0.292 | 0.245 | 0.155 | 0.135 | 0.079 | 0.065 | 0.061 | 0.045 | 0.033 |
| S-Rh 1 | 0.302 | 0.270 | 0.272 | 0.404 | 0.370 | 0.383 | 0.323 | 0.282 | 0.199 | 0.178 | 0.182 | 0.160 | 0.129 |
| R-Rh 1 | 0.029 | 0.072 | 0.082 | 0.151 | 0.180 | 0.222 | 0.211 | 0.231 | 0.186 | 0.191 | 0.204 | 0.194 | 0.158 |
| Rg 6 | 0.042 | 0.086 | 0.097 | 0.167 | 0.157 | 0.157 | 0.128 | 0.113 | 0.087 | 0.078 | 0.082 | 0.060 | 0.050 |
| F 4 | 0.086 | 0.188 | 0.224 | 0.454 | 0.494 | 0.583 | 0.541 | 0.539 | 0.409 | 0.398 | 0.401 | 0.352 | 0.269 |
| Rk 3 | 0.024 | 0.047 | 0.058 | 0.125 | 0.157 | 0.211 | 0.227 | 0.264 | 0.232 | 0.265 | 0.320 | 0.327 | 0.298 |
| Rh 4 | 0.034 | 0.068 | 0.086 | 0.194 | 0.255 | 0.358 | 0.398 | 0.474 | 0.438 | 0.513 | 0.633 | 0.661 | 0.608 |
| S-Rg 3 | 0.318 | 1.090 | 1.350 | 2.690 | 2.801 | 3.260 | 2.963 | 3.080 | 2.423 | 2.467 | 2.681 | 2.486 | 2.065 |
| R-Rg 3 | 0.214 | 0.653 | 1.097 | 2.212 | 2.275 | 2.890 | 2.296 | 2.833 | 2.180 | 2.172 | 3.145 | 2.938 | 2.594 |
| Rk 1 | 0.160 | 0.545 | 0.689 | 1.495 | 1.615 | 1.966 | 1.898 | 2.091 | 1.745 | 1.875 | 2.164 | 2.147 | 1.894 |
| Rg 5 | 0.216 | 0.740 | 0.938 | 2.082 | 2.276 | 2.812 | 2.752 | 3.089 | 2.658 | 2.913 | 3.380 | 3.446 | 3.039 |
| S-Rh 2 | 0.018 | 0.029 | 0.036 | 0.064 | 0.072 | 0.088 | 0.088 | 0.097 | 0.084 | 0.094 | 0.114 | 0.119 | 0.111 |
| R-Rh 2 | 0.017 | 0.022 | 0.030 | 0.048 | 0.052 | 0.069 | 0.060 | 0.066 | 0.052 | 0.054 | 0.070 | 0.068 | 0.062 |
| Rk 2 | 0.000 | 0.000 | 0.000 | 0.018 | 0.028 | 0.029 | 0.027 | 0.033 | 0.030 | 0.037 | 0.043 | 0.051 | 0.051 |
| Rh 3 | 0.000 | 0.007 | 0.010 | 0.036 | 0.047 | 0.064 | 0.065 | 0.080 | 0.078 | 0.093 | 0.120 | 0.135 | 0.136 |
| Rg 3 Total amount of | 0.532 | 1.743 | 2.447 | 4.902 | 5.076 | 6.150 | 5.259 | 5.913 | 4.603 | 4.639 | 5.826 | 5.424 | 4.659 |
| Rh 2 Total amount of | 0.035 | 0.051 | 0.066 | 0.112 | 0.124 | 0.157 | 0.148 | 0.163 | 0.136 | 0.148 | 0.184 | 0.187 | 0.173 |
| Rg 3 +Rh 2 | 0.567 | 1.794 | 2.513 | 5.014 | 5.200 | 6.307 | 5.407 | 6.076 | 4.739 | 4.787 | 6.010 | 5.611 | 4.832 |
| Rk 1 +Rg 5 | 0.376 | 1.285 | 1.627 | 3.577 | 3.891 | 4.778 | 4.650 | 5.180 | 4.403 | 4.788 | 5.544 | 5.593 | 4.933 |
| Rg 6 +F 4 | 0.128 | 0.274 | 0.321 | 0.621 | 0.651 | 0.740 | 0.669 | 0.652 | 0.496 | 0.476 | 0.483 | 0.412 | 0.319 |
| Total amount (%) | 1.669 | 4.106 | 5.252 | 10.500 | 11.070 | 13.335 | 12.131 | 13.406 | 10.879 | 11.393 | 13.599 | 13.189 | 11.497 |
Table 10 contents (%)% of 7 kinds of common ginsenosides in ginseng rootlet residue decocted at 125 ℃ under high temperature and high pressure
Table 11: content of 15 rare ginsenosides in ginseng rootlet dregs decocted at 130 ℃ under high temperature and high pressure (%)
| 20min | 40min | 1h | 1.5h | 2h | 2.5h | 10h | 12h | 14h | 16h | 20h | |
| S-Rg 2 | 0.162 | 0.212 | 0.243 | 0.267 | 0.273 | 0.290 | 0.183 | 0.250 | 0.197 | 0.173 | 0.097 |
| S-Rh 1 | 0.284 | 0.311 | 0.293 | 0.306 | 0.284 | 0.288 | 0.238 | 0.352 | 0.335 | 0.326 | 0.249 |
| R-Rh 1 | 0.028 | 0.026 | 0.035 | 0.047 | 0.054 | 0.061 | 0.107 | 0.191 | 0.198 | 0.227 | 0.201 |
| Rg 6 | 0.032 | 0.046 | 0.056 | 0.065 | 0.070 | 0.082 | 0.100 | 0.168 | 0.164 | 0.147 | 0.113 |
| F 4 | 0.058 | 0.089 | 0.109 | 0.130 | 0.139 | 0.161 | 0.291 | 0.524 | 0.564 | 0.599 | 0.529 |
| Rk 3 | 0.023 | 0.023 | 0.028 | 0.032 | 0.035 | 0.046 | 0.094 | 0.177 | 0.213 | 0.249 | 0.269 |
| Rh 4 | 0.032 | 0.034 | 0.042 | 0.049 | 0.053 | 0.068 | 0.146 | 0.292 | 0.359 | 0.430 | 0.490 |
| S-Rg 3 | 0.244 | 0.384 | 0.491 | 0.636 | 0.715 | 0.880 | 1.714 | 2.879 | 3.028 | 3.189 | 2.807 |
| R-Rg 3 | 0.172 | 0.247 | 0.438 | 0.592 | 0.671 | 0.687 | 2.186 | 2.684 | 2.812 | 3.193 | 2.832 |
| Rk 1 | 0.116 | 0.182 | 0.233 | 0.302 | 0.339 | 0.424 | 0.922 | 1.698 | 1.876 | 2.075 | 2.017 |
| Rg 5 | 0.166 | 0.248 | 0.307 | 0.411 | 0.453 | 0.575 | 1.304 | 2.416 | 2.699 | 2.999 | 3.002 |
| S-Rh 2 | 0.019 | 0.016 | 0.017 | 0.020 | 0.021 | 0.024 | 0.041 | 0.071 | 0.079 | 0.089 | 0.091 |
| R-Rh 2 | 0.005 | 0.016 | 0.020 | 0.021 | 0.021 | 0.022 | 0.041 | 0.060 | 0.063 | 0.069 | 0.063 |
| Rk 2 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.006 | 0.018 | 0.024 | 0.031 | 0.036 |
| Rh 3 | 0.000 | 0.000 | 0.003 | 0.003 | 0.004 | 0.006 | 0.022 | 0.042 | 0.052 | 0.064 | 0.076 |
| Rg 3 Total amount of | 0.416 | 0.631 | 0.929 | 1.228 | 1.386 | 1.567 | 3.900 | 5.563 | 5.840 | 6.382 | 5.639 |
| Rh 2 Total amount of | 0.024 | 0.032 | 0.037 | 0.041 | 0.042 | 0.046 | 0.082 | 0.131 | 0.142 | 0.158 | 0.154 |
| Rg 3 +Rh 2 | 0.440 | 0.663 | 0.966 | 1.269 | 1.428 | 1.613 | 3.982 | 5.694 | 5.982 | 6.540 | 5.793 |
| Rk 1 +Rg 5 | 0.282 | 0.430 | 0.540 | 0.713 | 0.792 | 0.999 | 2.226 | 4.114 | 4.575 | 5.074 | 5.019 |
| Rg 6 +F 4 | 0.090 | 0.135 | 0.165 | 0.195 | 0.209 | 0.243 | 0.391 | 0.692 | 0.728 | 0.746 | 0.642 |
| Total amount (%) | 1.341 | 1.834 | 2.315 | 2.881 | 3.132 | 3.614 | 7.395 | 11.822 | 12.663 | 13.860 | 12.872 |
Table 12 high temperature high pressure reaction of 7 kinds of common ginsenoside content in Leptoradix Ginseng residue at 130%
| 20min | 40min | 1h | 1.5h | 2h | 2.5h | 10h | 12h | 14h | 16h | 20h | |
| Rg 1 | 0.043 | 0.033 | 0.021 | 0.013 | 0.006 | 0.004 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Re | 0.166 | 0.127 | 0.080 | 0.050 | 0.027 | 0.016 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rb 1 | 0.687 | 0.646 | 0.465 | 0.440 | 0.273 | 0.212 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rc | 0.593 | 0.555 | 0.419 | 0.366 | 0.269 | 0.202 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rb 2 | 0.374 | 0.333 | 0.237 | 0.216 | 0.160 | 0.118 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rb 3 | 0.052 | 0.043 | 0.029 | 0.025 | 0.019 | 0.014 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 |
| Rd | 0.381 | 0.369 | 0.304 | 0.294 | 0.236 | 0.223 | 0.014 | 0.000 | 0.000 | 0.000 | 0.000 |
| Total amount (%) | 2.296 | 2.106 | 1.555 | 1.404 | 0.990 | 0.789 | 0.014 | 0.000 | 0.000 | 0.000 | 0.000 |
As can be seen from tables 5 to 8, the change rule of the common ginsenoside is similar to that of the common ginsenoside decocted at 100 ℃ and normal pressure when the common ginsenoside is decocted at high temperature and high pressure. With the progress of time, the content of the common ginsenoside in the water and the decoction dregs is gradually reduced, and the common ginsenoside can not be detected in the decoction dregs (44 hours) and the water liquid (48 hours). The rare ginsenoside changes in the way that the content of the rare ginsenoside in the water solution and the herb residue gradually increases from the beginning to the end. But the content of rare ginsenoside in the water solution begins to gradually decrease after the rare ginsenoside in the water solution increases to a certain degree, and the content of ginsenoside decreases to 0.703 percent after 48 hours; the content of rare ginsenoside in the medicine residue is continuously increased along with the prolonging of the decoction time, and reaches 12.94% after 40 hours, and then a rule of small fluctuation around the value is presented.
The same trend was observed at 120 deg.C, 125 deg.C, 130 deg.C, 135 deg.C, 140 deg.C and 145 deg.C thereafter, except that the time was shortened as the temperature was increased.
For example, as can be seen from tables 9-12, the contents of rare ginsenosides in the residue are gradually increased to a certain level and then basically remain unchanged, while the contents of common ginsenosides in the residue are gradually decreased to a final content of 0.
When the temperature is designed to be 150 ℃, the change rule of various ginsenosides is not changed, but the total amount of rare ginsenosides generated in the decoction dregs is obviously reduced, so the decoction temperature is not suitable to exceed 150 ℃.
As can be seen from FIGS. 7A to 10, the total amount of rare ginsenosides in the residue reaches a maximum value of 13% or more at 125 deg.C and 130 deg.C, and the optimal decoction time is 20 hours or more and 16 hours or so, respectively.
As can be seen from FIGS. 11 to 17, for rare ginsenoside Rg 3 The optimum temperature is 125 ℃ and 130 ℃, and the content of the slag can reach 6.15 percent and 6.38 percent; the optimal decoction time is 20 hr or more and 16 hr or so.
For rare ginsenoside Rk 1 And Rg 5 The optimum temperature is 125 deg.C and 145 deg.C, the content can respectively reach 5.59% and 5.6%, and the optimum decocting time is 40 hr or more and 5 hr or more.
Example 3: reducing Rg in water 3 Experimental condition screening of content
Under the conditions that the decoction time is 3,4,5 and hours at 120 ℃, 130 ℃, 135 ℃ and the liquid-material ratio is 7.5, the centrifugation immediately after the decoction is finished and the centrifugation of the next day are compared, and Rg in water is added after normal temperature feeding and heating to a higher temperature (60-100 ℃) 3 Influence of the content.
The experimental result shows that the Rg in the hot water is immediately centrifuged 3 The content is lower than that of the next day of centrifugation, and Rg in the decoction dregs 3 The content was increased with a difference of 0.3%. Heating to a higher temperature (60-100 deg.C) for feeding, and adding Rg in the residue at a temperature higher than normal temperature 3 The content of the sodium chloride is higher than that of the sodium chloride,the difference reaches about 0.1%.
Example 4: preparing extract from medicinal residue
Taking Leptoradix Ginseng, decocting at 115-145 deg.C with water temperature of 7.5:1 for 4-48 hr (corresponding to different decoction temperatures), centrifuging while hot, and discarding centrifugation water to obtain residue. Drying the residue, extracting with ethanol under reflux, recovering ethanol from the extractive solution under reduced pressure, and drying to obtain the desired extract. The HPLC method detects that the main components comprise 15 rare ginsenosides: 20 (S) -Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F4、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 、Rh 3 The total content of the ginsenoside reaches over 36 percent, and the ginsenoside Rg is basically not contained 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 、Rd。
The yield of the medicine residue is that the weight of the medicine residue/the weight of the American ginseng leaves is multiplied by 100 percent
The extract yield = extract weight/medicine residue weight × 100%
Table 13: decocting Leptoradix Ginseng to obtain residue and extract
Table 14: content (%) of rare ginsenoside in the extract:
| 145℃ | 140 |
135℃ | 130℃ | 125 |
120℃ | 115℃ | |
| S-Rg 2 | 0.238 | 0.600 | 0.853 | 0.361 | 0.747 | 1.193 | 0.894 |
| S-Rh 1 | 0.619 | 1.068 | 1.201 | 0.869 | 1.248 | 1.577 | 1.370 |
| R-Rh 1 | 0.576 | 0.637 | 0.587 | 0.741 | 0.736 | 0.727 | 0.665 |
| Rg 6 | 0.356 | 0.592 | 0.716 | 0.464 | 0.535 | 0.679 | 0.605 |
| F 4 | 1.541 | 1.889 | 1.752 | 1.662 | 1.771 | 1.816 | 1.767 |
| Rk 3 | 0.950 | 0.877 | 0.565 | 0.897 | 0.674 | 0.577 | 0.606 |
| Rh 4 | 1.424 | 1.167 | 0.826 | 1.471 | 1.092 | 0.906 | 1.070 |
| S-Rg 3 | 6.742 | 9.100 | 8.689 | 9.883 | 11.198 | 12.406 | 12.032 |
| R-Rg 3 | 7.955 | 8.567 | 7.402 | 9.882 | 9.739 | 10.167 | 9.220 |
| Rk 1 | 6.098 | 6.487 | 5.583 | 6.730 | 6.419 | 6.489 | 6.488 |
| Rg 5 | 8.786 | 8.985 | 7.600 | 9.772 | 9.103 | 9.201 | 9.573 |
| S-Rh 2 | 0.228 | 0.214 | 0.215 | 0.247 | 0.197 | 0.172 | 0.165 |
| R-Rh 2 | 0.210 | 0.186 | 0.133 | 0.154 | 0.132 | 0.121 | 0.075 |
| Rk 2 | 0.116 | 0.096 | 0.104 | 0.120 | 0.105 | 0.084 | 0.072 |
| Rh 3 | 0.232 | 0.149 | 0.177 | 0.245 | 0.183 | 0.151 | 0.133 |
| Rg 3 total amount of | 14.697 | 17.667 | 16.100 | 19.765 | 20.938 | 22.572 | 21.252 |
| Rh 2 Total amount of | 0.437 | 0.400 | 0.348 | 0.401 | 0.329 | 0.292 | 0.240 |
| Rg 3 +Rh 2 | 15.134 | 18.067 | 16.447 | 20.166 | 21.267 | 22.865 | 21.492 |
| Rk 1 +Rg 5 | 14.884 | 15.472 | 13.138 | 16.502 | 15.522 | 15.690 | 16.060 |
| Rg 6 +F 4 | 1.897 | 2.482 | 2.462 | 2.126 | 2.306 | 2.495 | 2.372 |
| Total amount (%) | 36.070 | 40.614 | 36.413 | 43.499 | 43.881 | 46.265 | 44.734 |
It can be seen that the total content of rare ginsenosides in the extract obtained by high temperature and high pressure decoction is between 30% and 50%, which is surprising. And at the temperature of 115-130 ℃, the total content of rare ginsenoside can reach more than 40%, and the total content of Rg3 can reach more than 14%, even more than 20%; the total content of Rk1 and Rg5 reaches more than 13 percent, even more than 15 percent. The extract with higher purity, the content of which reaches more than 50 percent and even more than 90 percent, is obtained by treating the extract with an organic solvent or by column chromatography.
Example 5: comparison of fresh Ginseng radix and dried Ginseng radix under normal pressure reflux and high temperature and high pressure
Heating dried Ginseng radix at 120 deg.C, 125 deg.C, 130 deg.C, and 135 deg.C for 5-16 hr at liquid-to-material ratio of 7.5. At this time, the water is rareThe ginsenoside content is very low, and Rg enriched in the medicine residue 3 F is 16 times of that in water 4 、Rk 3 、Rh 4 The content is also higher, and is more than 10 times of that in water liquid.
The trend of the obtained experimental results is the same as that of the above experiments by using fresh ginseng with 70% of water content as a raw material and decocting at high temperature and high pressure under the same experimental conditions. The content of common ginsenoside in water solution and residue is gradually reduced, and the content of rare ginsenoside in residue is gradually increased.
Only because of the fresh ginseng, the obtained medicine dregs have much lower content of rare ginsenoside than the dried ginseng.
Example 6: inhibiting effect of rare ginsenoside components with different ratios on cancer cells
100 mu L of A549 human lung cancer cell suspension is inoculated in a 96-well plate, and the inoculation density is 8000 cells/well. The 96-well cell culture plate was placed in a cell incubator (culture conditions 37 ℃,5% CO) 2 ) And culturing for 24h. Rare ginsenoside components with different proportions are prepared into high-concentration stock solutions by DMSO. Under the condition that the cells are in logarithmic growth, preparing a proper concentration gradient sample by using the stock solution, and co-culturing 24 groups of samples to be detected with different concentrations and the cells for 24 hours. For measuring the cell activity, cell supernatants of the respective groups of the 96-well plate were discarded, a medium containing 10% of the CCK-8 measuring reagent was prepared, 100. Mu.L of the medium was added to each reaction well, and the plate was placed in an incubator and incubated for about 1 hour. Absorbance at 450nm was measured with a microplate reader. Sample pair A549 tumor cell IC 50 The values are shown in the following table.
DLD1 human colon cancer cell suspension 100. Mu.L was seeded in 96-well plates at 8000 cells/well. The 96-well cell culture plate was placed in a cell incubator (culture conditions 37 ℃,5% CO) 2 ) And culturing for 24h. The rare ginsenoside components with different proportions are prepared into high-concentration stock solutions by DMSO. Under the condition that the cells are in logarithmic growth, preparing a proper concentration gradient sample by using the stock solution, and co-culturing 24 groups of samples to be detected with different concentrations and the cells for 24 hours. For measuring cell activity, cell supernatants of each group of 96-well plates were discarded, a medium containing 10% of CCK-8 detection reagent was prepared, 100. Mu.L of the medium was added to each reaction well, and the plate was placed in an incubator to incubateAbout 1 h. Absorbance at 450nm was measured with a microplate reader. Sample versus DLD1 tumor cell IC 50 The values are shown in the following table.
U251 human glioma cell suspension was seeded at 100 μ L in 96-well plates at 8000 cells/well. The 96-well cell culture plate was placed in a cell incubator (culture conditions 37 ℃,5% CO) 2 ) And culturing for 24h. The rare ginsenoside components with different proportions are prepared into high-concentration stock solutions by DMSO. Under the condition that the cells are in logarithmic growth, preparing a proper concentration gradient sample by using the stock solution, and co-culturing 24 groups of samples to be detected with different concentrations and the cells for 24 hours. For the measurement of cell activity, cell supernatants of each group of 96-well plates were discarded, a medium containing 10% of CCK-8 detection reagent was prepared, 100. Mu.L of the medium was added to each reaction well, and the plate was placed in an incubator and incubated for about 1 hour. Absorbance at 450nm was measured with a microplate reader. 24 groups of samples for U251 tumor cells IC 50 The values are shown in the following table.
TABLE 15 inhibition of cancer cells by different proportions of rare ginsenoside components
Through screening tests, a combination with better cancer cell inhibition activity in different rare ginsenoside combinations is screened out, and the combination shows synergistic antitumor activity on three kinds of cancer cells.
The applicant finds that the extract is also applicable to other active parts of ginseng and plants containing other ginsenosides, such as American ginseng, pseudo-ginseng and the like, and researches on ginseng leaves, pseudo-ginseng leaves, american ginseng and American ginseng leaves respectively through a large number of experiments, so that the same results are obtained, and a better extraction method is obtained, and the specific steps are as follows:
a method for preparing a product with high content of rare ginsenoside comprises any one of the following steps:
a: pulverizing folium Ginseng, adding appropriate amount of water, heating and refluxing under normal pressure for 36 hr or more, preferably 40 hr, 60 hr, 72 hr, 84 hr, 96 hr 108 hr or 120 hr or more, filtering, and drying the residue to obtain the desired product;
b: pulverizing folium Ginseng, adding appropriate amount of water, decocting at high temperature and under high pressure for 1 hr or more, 2 hr or more, 3 hr or more, 4 hr or more, 5 hr or more, preferably 6 hr or more, 8 hr or more, 10 hr or more, 12 hr or more, 14 hr or more, 16 hr or more, 18 hr or more, 20 hr or more, 22 hr or more, 24 hr or more, 28 hr or more, 30 hr or more, 32 hr or more, 34 hr or more, 36 hr or more, 38 hr or more, 40 hr or more, 42 hr or more, 44 hr or more, 48 hr or more, or 50 hr or more, filtering, and drying the residue to obtain the desired product.
A product for preparing high-content rare ginsenoside is prepared by any one of the following methods:
a: pulverizing Notoginseng radix, adding appropriate amount of water, heating and refluxing under normal pressure for more than 48 hr, preferably 60 hr, 72 hr, 84 hr, 96 hr 108 hr or more than 120 hr, filtering, and drying the residue to obtain the desired product;
b: pulverizing Notoginseng radix, adding appropriate amount of water, decocting at high temperature and under high pressure for 4 hr or more, preferably 6 hr or more, 8 hr or more, 10 hr or more, 12 hr or more, 14 hr or more, 16 hr or more, 18 hr or more, 20 hr or more, 22 hr or more, 24 hr or more, 28 hr or more, 30 hr or more, 32 hr or more, 34 hr or more, 36 hr or more, 38 hr or more, 40 hr or more, 42 hr or more, 44 hr or more, 48 hr or more, or 50 hr or more, filtering, and drying the residue to obtain the desired product.
A method for preparing a product with high content of rare ginsenoside comprises any one of the following steps:
a: drying and pulverizing folium Notoginseng, adding appropriate amount of water, heating and refluxing under normal pressure for more than 48 hr, preferably 60 hr, 72 hr, 84 hr, 96 hr 108 hr or more than 120 hr, filtering, and drying the residue to obtain the desired product;
b: drying and pulverizing folium Notoginseng, adding appropriate amount of water, decocting at high temperature and under high pressure for more than 2 hr, preferably more than 4 hr, preferably more than 6 hr, more than 8 hr, more than 10 hr, more than 12 hr, more than 14 hr, more than 16 hr, more than 18 hr, more than 20 hr, more than 22 hr, more than 24 hr, more than 28 hr, more than 30 hr, more than 32 hr, more than 34 hr, more than 36 hr, more than 38 hr, more than 40 hr, more than 42 hr, more than 44 hr, more than 48 hr or more than 50 hr, filtering, and drying the residue to obtain the desired product.
A product for preparing high-content rare ginsenoside is prepared by any one of the following methods:
a: pulverizing radix Panacis Quinquefolii, adding appropriate amount of water, heating and refluxing under normal pressure for more than 48 hr, preferably 60 hr, 72 hr, 84 hr, 96 hr or 108 hr, filtering, and drying the residue to obtain the desired product;
b: drying and pulverizing radix Panacis Quinquefolii, adding appropriate amount of water, decocting at high temperature and high pressure for more than 4 hr, preferably more than 6 hr, more than 8 hr, more than 10 hr, more than 12 hr, more than 14 hr, more than 16 hr, more than 18 hr, more than 20 hr, more than 22 hr, more than 24 hr, more than 28 hr, more than 30 hr, more than 32 hr, more than 34 hr, more than 36 hr, more than 38 hr, more than 40 hr, more than 42 hr, more than 44 hr, more than 48 hr or more than 50 hr, filtering, and drying the residue to obtain the desired product.
A method for preparing a product with high content of rare ginsenoside comprises any one of the following steps:
a: drying and pulverizing folium Panacis Quinquefolii, adding appropriate amount of water, and heating and refluxing under normal pressure for more than 48 hr, preferably 60 hr, 72 hr, 84 hr, 96 hr or more than 108 hr; more preferably 90 hours; filtering, and drying the residues to obtain the required product;
b: drying and pulverizing leaves of Panax quinquefolium, adding appropriate amount of water, decocting at high temperature and under high pressure for 2 hr or more than 4 hr, preferably 6 hr or more, 8 hr or more, 10 hr or more, 12 hr or more, 14 hr or more, 16 hr or more, 18 hr or more, 20 hr or more, 22 hr or more, 24 hr or more, 28 hr or more, 30 hr or more, 32 hr or more, 34 hr or more, 36 hr or more, 38 hr or more, 40 hr or more, 42 hr or more, 44 hr or more, 48 hr or more, or 50 hr or more, filtering, and drying the residue to obtain the desired product.
Because the experimental data are too huge, the applicant carries out divisional application on different parts and different plants, and detailed data are shown in patent applications on the same day.
The above description is a general description of the invention. Changes in form and substitution of equivalents may be made as circumstances or practical, and although specific terms are employed herein, they are intended in a descriptive sense and not for purposes of limitation. Various changes or modifications may be effected by one skilled in the art and these equivalents are intended to be within the scope of the invention as defined in the claims appended hereto.
Claims (12)
1. A product prepared from Ginseng radix contains high content of rare ginsenoside, and does not contain common ginsenoside; wherein the rare ginsenoside comprises: 20 (S) -Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 、Rh 3 And the common ginsenoside comprises: rg (Rg) 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 、Rd；
The preparation method comprises the following steps:
a: pulverizing Ginseng radix, adding appropriate amount of water, heating and refluxing under normal pressure for more than 48 hr, filtering, and drying the residue to obtain the desired product;
b: pulverizing Ginseng radix, adding appropriate amount of water, decocting at high temperature under high pressure for more than 4 hr, filtering, and drying the residue to obtain the desired product; the high temperature and high pressure are not lower than 110 ℃;
in the preparation method A, the liquid-material ratio is (20-9): 1;
in the preparation method B, the liquid-material ratio is (25-6): 1.
2. The product of claim 1, wherein the total content of rare ginsenosides is not less than 5%, and the total content of common ginsenosides is less than 1%.
3. A product prepared from ginseng as claimed in claim 1 or 2, wherein:
in preparation method A, heating and refluxing are carried out for 60 hours, 72 hours, 84 hours, 96 hours, 108 hours or more than 120 hours under normal pressure;
in preparation method B, decocting at high temperature and high pressure for 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 28 hours, 30 hours, 32 hours, 34 hours, 36 hours, 38 hours, 40 hours, 42 hours, 44 hours, 48 hours, or more than 50 hours; the high temperature and high pressure is not lower than 115 ℃,120 ℃, 125 ℃, 130 ℃, 135 ℃,140 ℃, 145 ℃ or 150 ℃;
in the preparation method A, the liquid-material ratio is (15-10) 1 or 10;
in preparation method B, the liquid-material ratio is (20-7): 1, (10-8): 1, or 7.5.
4. A method for preparing a high-content rare ginsenoside product is characterized in that the preparation method comprises the following steps:
a: pulverizing Ginseng radix, adding appropriate amount of water, heating and refluxing under normal pressure for more than 48 hr, filtering, and drying the residue to obtain the desired product;
b: pulverizing Ginseng radix, adding appropriate amount of water, decocting at high temperature under high pressure for more than 4 hr, filtering, and drying the residue to obtain the desired product; the high temperature and high pressure is not lower than 110 ℃;
in the preparation method A, the liquid-material ratio is (20-9) to 1;
in the preparation method B, the liquid-material ratio is (25-6): 1.
5. The process according to claim 4, wherein:
in the preparation method A, the heating reflux is carried out for 60 hours, 72 hours, 84 hours, 96 hours, 108 hours or more than 120 hours under normal pressure;
in preparation method B, decocting at high temperature and high pressure for 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, 24 hours, 28 hours, 30 hours, 32 hours, 34 hours, 36 hours, 38 hours, 40 hours, 42 hours, 44 hours, 48 hours, or more than 50 hours; the high temperature and high pressure is not lower than 115 ℃,120 ℃, 125 ℃, 130 ℃, 135 ℃,140 ℃, 145 ℃ or 150 ℃;
in the preparation method A, the liquid-material ratio is (15-10) 1 or 10;
in preparation method B, the liquid-material ratio is (20-7): 1, (10-8): 1, or 7.5.
6. The process according to claim 4 or 5, wherein: further extracting the residue with organic solvent or aqueous organic solvent to obtain extract with high content of rare ginsenoside.
7. The process according to claim 4 or 5, wherein:
the medicinal residue contains one or more of 15 rare ginsenosides, specifically 20 (S) -Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 、Rh 3 Substantially free of common ginsenoside including Rg 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 、Rd；
The total content of rare ginsenoside in the residue is not less than 5%, and the total content of common ginsenoside is not more than 1%.
8. The process according to claim 6, wherein:
the extract contains one or more of 15 rare ginsenosides, specifically 20 (S) -Rg 2 、20(S)-Rh 1 、20(R)-Rh 1 、Rg 6 、F 4 、Rk 3 、Rh 4 、20(S)-Rg 3 、20(R)-Rg 3 、Rk 1 、Rg 5 、20(S)-Rh 2 、20(R)-Rh 2 、Rk 2 、Rh 3 Substantially free of common ginsenoside including Rg 1 、Re、Rb 1 、Rc、Rb 2 、Rb 3 、Rd；
The total content of rare ginsenoside in the extract exceeds 20%, and the total content of common ginsenoside is less than 5%.
9. An anti-tumor composition, wherein the active component of the composition is the composition of any one of the following components:
from Rg 3 And Rg 5 The weight ratio of the composition is (4-6) to (1-3); or (7-11.5) to (3.5-5);
from F 4 And Rh 2 A composition consisting of, by weight, 10;
from Rh 4 And Rh 2 The composition comprises (1-10) by weight: (10-1).
10. The composition of claim 9, wherein the tumor is lung cancer, colon cancer or glioma.
11. Use of a product according to any one of claims 1 to 3 or a composition according to claim 9 for the preparation of an anti-tumour product.
12. The use according to claim 11, wherein the tumor is lung cancer, colon cancer or glioma.
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