Preparation method and application of American ginseng extract rich in rare ginsenosides

Abstract

The present invention discloses a preparation method and application of an American ginseng extract rich in rare ginsenosides, belonging to the technical field of rare ginsenoside preparation. The method solves the problem that the content of Rg3, Rg5, and Rk1 in existing extracts needs to be improved. First, the root of American ginseng is sliced ​​or crushed and mixed with water, and boiled for 2-5 hours; (2) a 50%-70% ethanol solution is added at 40-80°C for soaking and filtering, and the filter cake is further added with a 50%-70% ethanol solution at 40-80°C for soaking and filtering, the filtrate is combined, heated at 120-140°C for concentration, and ethanol is recovered; (3) the concentrate is applied to a macroporous resin column, first rinsed with 3 column volumes of water, then eluted with 60-95% ethanol, the alcohol phase eluate is combined, heated at 120-135°C for concentration, and ethanol is recovered to obtain an American ginseng extract rich in rare ginsenosides Rg3, Rg5, and Rk1, which can be used in functional foods, health products, and biopharmaceutical products.

Description

Technical Field

[0001] The present invention relates to the technical field of rare ginsenoside preparation, and in particular to a method for preparing an American ginseng extract rich in rare ginsenosides Rg3, Rg5 and Rk1. Background Art

[0002] American ginseng contains ginsenosides, amino acids, polysaccharides, volatile oils, vitamins, and organic acids. Ginsenosides are considered the most representative active ingredients in American ginseng. Numerous studies have shown that rare ginsenosides such as Rg3, Rg5, Rk1, and Rh2, obtained through structural modification of ginsenosides, exhibit enhanced pharmacological activity. Steaming is a traditional method for preparing rare ginsenosides Rg5 and Rk1. This involves steaming and drying American ginseng (Araliaceae) plants such as Panax quinquefolium at a constant temperature for extended periods of time, transforming the primary protopanaxadiol group of ginsenosides, Rb1, Rb2, Rb3, Rc, and Rd, into the rare ginsenosides Rg5 and Rk1. For example, patent application number CN201810333188 discloses a method for preparing low-polarity rare ginsenosides Rg5 / Rk1 and Rh3 / Rk2. It studies a method for preparing ginsenosides Rg5 / Rk1, etc. from the stem and leaf powder of Araliaceae plants through high-temperature steaming, soaking extraction, adding sodium hydroxide, etc., and reacting with oxygen. This method uses various chemical alkaline reagents as catalysts. The preparation efficiency of ginsenoside Rg5 is high, but there is certain environmental pollution. Moreover, due to the addition of chemical agents such as sodium hydroxide, consumers are worried that the residues of chemical agents will affect food safety. At the same time, the addition of chemical agents will also affect the taste of the product. Most importantly, relevant laws and regulations have clear provisions on the use of sodium hydroxide in food, and the sodium hydroxide content must be completely removed, which further increases the difficulty of subsequent processing. Summary of the Invention

[0003] The purpose of the present invention is to provide a method for preparing an American ginseng extract rich in rare ginsenosides, which solves the problem of the use of chemical agents in the existing methods for preparing rare ginsenosides, resulting in increased difficulty in post-processing and difficulty in consumer acceptance, and at the same time solves the problem that the content of Rg3, Rg5, and Rk1 in the existing extracts needs to be improved. The present invention utilizes the components contained in American ginseng itself as structural modifiers and uses American ginseng roots as starting materials to develop a method for preparing an American ginseng extract rich in rare ginsenosides Rg3, Rg5, and Rk1 based on traditional methods. This method does not require the addition of other acidic and alkaline chemicals or other plant acids, avoids the need for high-temperature and high-pressure steaming, has high extraction efficiency, retains the nutritional components of American ginseng, is low-cost, green and environmentally friendly, and can be eaten directly.

[0004] In order to achieve the above-mentioned purpose, the present invention adopts the following technical scheme: A preparation method of American ginseng extract rich in rare ginsenosides, comprising the following steps: (1) slicing or crushing American ginseng roots, mixing with water, and boiling for 2-5 hours; (2) adding 50%-70% Vol raw liquor at 40-80℃ and filtering, adding 50%-70% Vol raw liquor to the filter cake, soaking it at 40-80℃ and filtering, combining the filtrate, and heating and concentrating at 120-140℃; (3) applying the concentrate to a macroporous resin column, first washing with water, and then eluting with 60-95% ethanol, combining the alcohol phase eluate, and heating and concentrating at 120-135℃ to obtain an American ginseng extract rich in rare ginsenosides Rg3, Rg5, and Rk1.

[0005] In step (1), the mass volume ratio of American ginseng root to water is 1:2-1:5 kg / L, and the boiling temperature is 80-100°C.

[0006] In step (2), the mass volume ratio of American ginseng root to 50%-70% Vol puree wine is 1:10-1:30 kg / L, and the soaking time is 3-8 hours.

[0007] In the American ginseng extract extracted by the above method, the total ginsenoside content is greater than 50 wt.%, the Rg3 content is greater than 12 wt.%, the Rk1 content is greater than 6 wt.%, and the Rg5 content is greater than 9 wt.%.

[0008] Compared with the prior art, the present invention has the following beneficial effects: (1) Based on the characteristics of American ginseng as a plant with medicinal and edible properties, a mild and natural traditional food processing method is used to process and extract an extract rich in rare ginsenosides. According to the processing requirements for the consumption of American ginseng in the medicinal and edible properties, the extract can be directly used in food production; (2) The extraction process uses raw wine instead of conventional ethanol reagent, which not only reduces the use of ethanol reagent but also increases the content of rare ginsenosides Rg3, Rg5, and Rk1 in the extract; (3) The concentration temperature is increased during the concentration process, which accelerates the removal of ethanol, further promotes the conversion of proto-ginsenosides of the American ginseng into rare ginsenosides, and increases the content of Rg3, Rg5, and Rk1. BRIEF DESCRIPTION OF THE DRAWINGS

[0009] Figure 1 This is the liquid chromatogram of the American ginseng extract in Example 1.

[0010] Figure 2 This is the liquid chromatogram of the American ginseng extract in Example 2.

[0011] Figure 3 This is the liquid chromatogram of the American ginseng extract in Comparative Example 3. DETAILED DESCRIPTION

[0012] The specific implementation of the present invention is further described below in conjunction with the accompanying drawings and specific technical solutions.

[0013] The present invention discloses a method for preparing rare ginsenosides Rg3, Rg5 and Rk1 by using a traditional method. The method can prepare rare ginsenosides efficiently and greenly, and solves the problem that strong acid, strong base or other plant acids are additionally introduced into the existing preparation process, resulting in that the American ginseng extract contains substances other than American ginseng, does not meet the national preparation requirements for the medicinal and edible properties of American ginseng, and cannot be directly used in ordinary food.

[0014] The method for preparing a ginseng extract rich in rare ginsenosides of the present invention comprises the following steps: (1) slicing or crushing the root of ginseng and mixing it with water in a mass-to-volume ratio of 1:2-1:5 (kg / L), and boiling it at 80-100°C for 2-5 hours; (2) adding 50%-70% Vol raw liquor in a mass-to-volume ratio of 1:10-1:30 (kg / L), soaking it at 40-80°C for 3-8 hours, filtering it, and continuing to add mass volume to the filter cake. The concentrate is loaded onto a macroporous resin column, first rinsed with 3 column volumes of water, then eluted with 60-95% ethanol, the alcohol phase eluates are combined, heated and concentrated at 120-135°C, and the ethanol is recovered to obtain an American ginseng extract rich in rare ginsenosides Rg3, Rg5, and Rk1.

[0015] The American ginseng extract extracted by the above method contains over 50% total saponins, primarily including Rb1, Rg3, Rk1, and Rg5. Rb1 is less than 16.5% by weight, Rg3 is greater than 12% by weight, Rk1 is greater than 6% by weight, and Rg5 is greater than 9% by weight. Rb1 is the prototype ginsenoside in American ginseng and is edible directly. Rg3, Rk1, and Rg5 are valuable rare ginsenosides, and numerous studies have demonstrated their efficacy in treating conditions such as chronic heart failure, stable angina pectoris due to coronary heart disease, arrhythmias, diabetes and its complications, hyperlipidemia, fatty liver disease, Alzheimer's disease, dysmenorrhea, metabolic syndrome, and tumors. These extracts possess medicinal and health benefits, and therefore can be used in functional foods, health supplements, and biopharmaceuticals.

[0016] The masses in steps (1) and (2) refer to the mass of American ginseng root in step (1).

[0017] In step (1), the boiling temperature is preferably 90-95° C. In one or more embodiments, the boiling temperature in step (1) is 80° C., 85° C., 90° C., 95° C., 100° C., or a range defined by any two of the above values.

[0018] In step (2), the raw liquor refers to solid-state liquor. The alcohol content of the raw liquor is 50%-70%. In one or more embodiments, the alcohol content of the raw liquor is 50%, 60%, 70%, or a range defined by any two of the above values.

[0019] Existing rare ginsenosides are extracted with alcohol and then heated and concentrated to remove ethanol. The heating temperature is lower than the boiling point of ethanol (78°C). For example, the rotary evaporation concentration temperature is as low as 40°C, which only plays a role in concentration. In steps (2) and (3) of the present invention, heating and concentrating at 120-140°C can quickly remove the solvent. More importantly, it further enhances the conversion of the prototype ginsenoside Rb1 contained in American ginseng to Rg3, and the conversion of Rg3 to Rg5 and Rk1, thereby increasing the content of rare ginsenosides. In one or more embodiments, the heating and concentration temperature in step (2) is 120°C, 125°C, 130°C, 135°C, 140°C, or a range defined by any two of the above values. The heating and concentration temperature in step (3) is 120°C, 125°C, 130°C, 135°C, or a range defined by any two of the above values.

[0020] The purpose of applying the concentrate to a macroporous resin column in step (3) is to remove substances such as polysaccharides and further purify the ginsenosides in the extract.

[0021] Example 1

[0022] (1) Add 20g of American ginseng root slices to 80ml of water and boil at 90-95℃ for 4 hours.

[0023] (2) Add 400 ml of 60% Vol original liquor, soak at 75-80°C for 5 hours and filter, add 400 ml of 60% Vol original liquor to the filter cake, soak at 75-80°C for 3 hours and filter, combine the filtrate, heat at 125°C and concentrate, the 60% Vol original liquor is Yufang Kaoliang original liquor (produced by Xinqiang Liquor Co., Ltd., Qionglai City, Sichuan Province).

[0024] (3) The concentrate was loaded onto a macroporous resin column HP20, first rinsed with 3 column volumes of water, then eluted with 80% ethanol, the alcohol phase eluates were combined, heated at 125°C for concentration, and the ethanol was recovered to obtain the American ginseng extract.

[0025] Comparative Example 1

[0026] This comparative example is the same as Example 1 except that 60% Vol of raw liquor in step (2) is replaced by 60% ethanol.

[0027] Comparative Example 2

[0028] This comparative example is the same as Example 1 except that the concentration is heated at 60°C in steps (2) and (3).

[0029] The HPLC external standard method was used to detect the American ginseng extract in Example 1-3. The liquid chromatograms are shown in FIG. Figure 1-3 The specific composition and content are shown in Table 1. It can be seen from Table 1 that compared with the use of ethanol, the use of raw wine in step (2) can increase the content of Rg3, Rg5 and Rk1. This may be because the addition of raw wine promotes the conversion of protopanaxadiol-type ginsenosides Rb1 and Rb2 to Rg3(S) and Rg3(R), and also promotes the conversion of Rg3(S) and Rg3(R) to Rg5 and Rk1.

[0030] Table 1: Ginsenoside content in American ginseng extract (wt.%)

[0031] name Example 1 Comparative Example 1 Comparative Example 2 Re 0 0.224 0.837 Rg1 0.136 0 8.49 Rb1 16.437 20.969 33.925 Rg6 0.749 0 0.144 F4 3.547 3.632 0 Rg3 (S) 6.451 6.12 0 Rg3 (R) 6.4 5.98 0.795 Rk3 0.172 0 0.047 Rk1 6.923 5.884 0.249 Rg5 9.804 8.714 0.792 Total ginsenosides detected 50.619 51.523 45.279 (Rk1+Rg3+Rg5) / total ginsenosides 58.4 51.8 4

[0032] HPLC detection conditions are as follows: chromatographic column: octadecyl bonded silica gel column or similar type chromatographic column (250×4.6mm, 5m); mobile phase: phase A: acetonitrile; phase B: water; flow rate: 1.0mL / min; column temperature: 30℃; detection wavelength: 202nm; injection volume: 10μL; gradient: gradient conditions are shown in Table 2.

[0033] Table 2

[0034] time Mobile phase A (%, volume ratio) Mobile phase B (% by volume) 0～0.1 25% 75% 0.1～5 25%~30% 75%~70% 5～14 30%~38% 70%~62% 14～28 38%~38% 62%~62% 28～30 38%~46% 62%~54% 30～50 46% ~74% 54%~26% 50～60 74%~100% 26% ~ 0% 60～65 100%~25% 0% ~75% .

Claims

1. A method for preparing an American ginseng extract rich in rare ginsenosides, characterized in that: The following steps are involved: (1) Slice or crush the root of American ginseng, mix it with water, and boil it; (2) add 50%-70% Vol of raw liquor, soak it at 40-80℃ and filter it, add 50%-70% Vol of raw liquor to the filter cake, soak it at 40-80℃ and filter it, combine the filtrate, and heat and concentrate it at 120-140℃; (3) Put the concentrate on a macroporous resin column HP20, first rinse it with water, then elute it with 60-95% ethanol, combine the alcohol phase eluate, and heat and concentrate it at 120-135℃ to obtain an American ginseng extract rich in rare ginsenosides Rg3, Rg5, and Rk1, wherein the content of Rg3 in the total saponins is greater than 12 wt.%, the content of Rk1 in the total saponins is greater than 6 wt.%, and the content of Rg5 in the total saponins is greater than 9 wt.%.

2. The method for preparing the American ginseng extract rich in rare ginsenosides according to claim 1, characterized in that: In step (1), the mass volume ratio of American ginseng root to water is 1:2-1:5 kg / L, the boiling temperature is 80-100° C., and the boiling time is 2-5 hours.

3. The method for preparing the American ginseng extract rich in rare ginsenosides according to claim 1, characterized in that: In step (2), the mass volume ratio of American ginseng root to 50%-70% Vol puree wine is 1:10-1:30 kg / L, and the soaking time is 3-8 hours.

4. The method for preparing the American ginseng extract rich in rare ginsenosides according to claim 1, characterized in that: In step (2), the raw liquor refers to liquor prepared by solid-state method.

5. An American ginseng extract rich in rare ginsenosides prepared by the method according to any one of claims 1 to 4.

6. Use of the American ginseng extract rich in rare ginsenosides according to claim 5 in the preparation of health products and biopharmaceutical products.

Images