阅读说明：本技术 抗衰老的中药提取物及其提取方法和用途 (Anti-aging traditional Chinese medicine extract and extraction method and application thereof ) 是由 陈谨 何云燕 苏俊 李艾 于 2020-08-04 设计创作，主要内容包括：本发明公开了抗衰老的中药提取物及其提取方法和用途,本发明人参黄芪提取物中含有黄芪甲苷20～30％、人参皂苷Rg1 1～5％、人参皂苷Re 2～6％、人参皂苷Rb1 0.2～0.5％,高效液相色谱指纹图谱中包括17个特征峰,可有效提高端粒酶的活性,延缓细胞衰老,可应用于制备抗衰老产品。(The invention discloses an anti-aging traditional Chinese medicine extract, an extraction method and application thereof, wherein the ginseng astragalus extract contains 20-30% of astragaloside IV, 78-5% of ginsenoside Rg 11, 2-6% of ginsenoside Re and 10.2-0.5% of ginsenoside Rb, and a high performance liquid chromatography fingerprint comprises 17 characteristic peaks, so that the activity of telomerase can be effectively improved, the cell aging can be delayed, and the extract can be applied to preparation of anti-aging products.)

1. The composition is characterized by comprising the following components in parts by weight: 20-30 parts of astragaloside IV, 5-5 parts of ginsenoside Rg 11, 2-6 parts of ginsenoside Re and 10.2-0.5 part of ginsenoside Rb; further, the composition comprises the following components in parts by weight: 22-28 parts of astragaloside IV, 11.8-4 parts of ginsenoside Rg, 3-5 parts of ginsenoside Re and 10.2-0.3 part of ginsenoside Rb.

2. The extract is characterized by comprising the following raw materials in parts by weight: 1 part of ginseng and/or American ginseng, 4-8 parts of astragalus, and further 4-6 parts of astragalus; the extract contains 20-30% of astragaloside IV, 20-5% of ginsenoside Rg 11, 2-6% of ginsenoside Re and 10.1-0.5% of ginsenoside Rb; further, the extract contains 22-28% of astragaloside IV, 11.8-4% of ginsenoside Rg, 3-5% of ginsenoside Re and 10.2-0.3% of ginsenoside Rb;

further, the total saponin content in the extract is 75-95%, and the total polysaccharide content is 3-10%; furthermore, the content of the total saponins is 80-90%, and the content of the total polysaccharides is 5-8%.

3. The extract according to claim 2, wherein the ginseng is ginseng and/or a ginseng processed product; further, the processed ginseng product is selected from one or more of sun-dried ginseng, red ginseng and active ginseng.

4. An extract comprising ginsenoside Re; the high performance liquid chromatography fingerprint spectrum of the extract at least comprises 6 characteristic peaks, the characteristic peak of the ginsenoside Re is taken as a reference peak, and the relative retention time of the 6 characteristic peaks and the reference peak is as follows: 0.95, 1.00, 1.80, 2.13, 2.36, 3.20; further, the method also comprises 11 characteristic peaks, and the relative retention time of the 11 characteristic peaks and a reference peak is as follows: 1.66, 1.69, 1.73, 1.86, 1.88, 1.94, 1.99, 2.02, 2.28, 2.40, 2.45;

further, the raw materials comprise the following raw materials in parts by weight: 1 part of ginseng and/or American ginseng, 4-8 parts of astragalus, and further 4-6 parts of astragalus.

5. The extract according to claim 4, wherein the ginseng is ginseng and/or a ginseng processed product; further, the processed ginseng product is selected from one or more of sun-dried ginseng, red ginseng and active ginseng.

6. Extract according to claim 4, characterized in that the relative peak areas of the characteristic peaks with relative retention times of 0.95, 1.00, 1.80, 2.13, 2.36 and 3.20 are, in order: 48.41%, 100.00%, 107.04%, 48.70%, 340.51% and 54.83%; further, the relative peak areas of the characteristic peaks with relative retention times of 1.66, 1.69, 1.73, 1.86, 1.88, 1.94, 1.99, 2.02, 2.28, 2.40 and 2.45 are, in order: 9.52%, 17.21%, 2.61%, 4.16%, 22.88%, 10.92%, 16.34%, 8.33%, 3.37%, 5.14% and 14.59%.

7. The method for detecting the high performance liquid fingerprint of the extract as claimed in any one of claims 2, 4 and 6, wherein the qualitative and/or quantitative detection is carried out by high performance liquid chromatography, and the chromatographic conditions comprise:

a chromatographic column: c18 silica gel column;

mobile phase: water and acetonitrile, using the following gradient elution procedure:

time (min) Water (V%) Acetonitrile (V%) 0 80～90 10～20 30～35 60～70 30～40 70～85 10～30 70～90

Further, the following elution procedure was employed:

time (min) Water (V%) Acetonitrile (V%) 0 85 16 33 66 34 78.5 20 80

。

8. The detection method according to claim 7, characterized by further comprising the condition of:

specification of chromatographic column: 150 x 3.9mm, 5 μm;

column temperature: 25-35 ℃, preferably 30 ℃;

flow velocity: 0.8-1.2 mL/min, preferably 1.00 mL/min;

detecting wavelength: 200-205 nm, preferably 203 nm.

9. A preparation method of a natural plant extract is characterized by comprising the following steps: (1) extracting the raw materials with water; (2) enriching by adopting macroporous adsorption resin; (3) carrying out enzymolysis; (4) performing membrane filtration, wherein the molecular weight is intercepted by 200-1000 Da; (5) solid-liquid separation;

the plant is selected from one or two of ginseng and American ginseng, and/or astragalus; further, 1 part of ginseng and/or American ginseng, 4-8 parts of astragalus, and further 4-6 parts of astragalus.

10. The method according to claim 9, wherein the ginseng is ginseng and/or a ginseng processed product; further, the processed ginseng product is selected from one or more of sun-dried ginseng, red ginseng and active ginseng.

11. The method for preparing a compound of claim 9, wherein in the water extraction step, the ratio of water: the liquid-material ratio of the raw materials is 5-15: 1 mL/g;

further, the number of times of water extraction is 1-4; further, it is 2 times.

12. The preparation method according to claim 9, wherein the macroporous adsorption resin enrichment is to adsorb an extracting solution obtained by water extraction by using macroporous adsorption resin, sequentially wash the extracting solution by using water, 10-40% ethanol and 60-80% ethanol, and collect a part obtained by washing the extracting solution by using 60-80% ethanol;

further, washing with water, 25-30% ethanol and 70-80% ethanol respectively, and collecting the parts obtained by washing with 70-80% ethanol;

further, the macroporous adsorption resin is nonpolar or weak polar macroporous adsorption resin; preferably, the macroporous adsorption resin is selected from D-101, HPD-10, X-5 and AB-8.

13. The extraction method according to claim 9, wherein the enzymolysis is carried out on the substance obtained after the macroporous adsorption resin is enriched; the enzyme is a complex enzyme consisting of beta-glucosidase and xylosidase in a weight ratio of 2-4: 1;

further, the enzymolysis conditions are as follows: performing enzymolysis on the raw materials at the conditions of a complex enzyme with the mass of 0.2-1.0% and the pH of 5-6 and the temperature of 40-55 ℃;

further, the using amount of the complex enzyme is 0.4-0.6%;

further, the enzymolysis time is 3-6 h; further, the enzymolysis time is 4 h.

14. The extract prepared by the method of any one of claims 9 and 11 to 13, wherein the extract comprises one of the following extracts:

(1) a ginseng extract; (2) american ginseng extract; (3) an extract of Astragalus membranaceus; (4) a mixed extract of ginseng and American ginseng; (5) a mixed extract of ginseng and astragalus; (6) a mixed extract of American ginseng and astragalus; (7) mixed extracts of Ginseng radix, radix Panacis Quinquefolii and radix astragali.

15. The composition is characterized by comprising the following raw materials in parts by weight: 1 part of the ginseng extract and/or the American ginseng extract as claimed in claim 14, and 5-14 parts of the astragalus extract as claimed in claim 14; further, the composition comprises 1 part of the ginseng extract and/or the American ginseng extract of claim 14 and 5.5-11 parts of the astragalus extract of claim 11.

16. Use of one or more of the extract of claims 2, 4, 6, the extraction process of claims 9, 11-13, the extract of claim 14, the composition of claim 1 or 15, in the preparation of a product for increasing telomerase activity and/or increasing telomere length;

further, the product is telomerase activator;

further, the product is an anti-aging product;

further, the product is a product for delaying cell aging.

17. A product comprising one or more of the extract of claims 2, 4, 6, the extract of claim 14, the composition of claim 1 or 15.

Technical Field

The invention relates to the field of traditional Chinese medicine extracts and extraction methods, in particular to an anti-aging traditional Chinese medicine extract and an extraction method and application thereof.

Background

Cells are the basic unit of organism structure and function, and are also the basic unit of organism aging. Cell senescence (aging) refers to a process in which the proliferation and differentiation ability and physiological functions of cells gradually decline with the passage of time during the course of performing life activities. The life cycle of a cell goes through several stages of undifferentiated, differentiated, growth, maturation, senescence and death.

Although death from aging is an inevitable natural rule, delaying aging, especially in an effort to avoid pathological aging, is possible. The population of the elderly, over 60 years of age, reported to reach 20 billion in 2050 worldwide, approximately 20-30% of the general population. The method faces the trend of accelerating the aging process of population and generally improving the service life of population, ensures that the old people enjoy good health and higher living quality and becomes a major problem of common attention of social science and life science, deeply researches more effective anti-aging products and is certainly beneficial to realizing the aim of 'healthy aging' in China. Therefore, the research on delaying senility and the development of products for delaying senility have important scientific significance and social value.

Disclosure of Invention

For a long time, people are constantly exploring methods for delaying senescence and prolonging life, the research on prolonging life by traditional Chinese medicine has a long history, precious literature records and abundant practical experience, and also records that many traditional Chinese medicinal materials have anti-aging effects, and in the research and development process of anti-aging products, researchers find that although many traditional Chinese medicinal materials have anti-aging effects, the anti-aging effects are not very ideal or remarkable, and even if an extract with higher active substance content is adopted, the anti-aging activity of the traditional Chinese medicinal materials is still difficult to further improve.

Therefore, if the anti-aging activity of the traditional Chinese medicinal materials is further improved, an extract capable of effectively delaying aging is obtained, which is a problem to be solved in the field.

The invention aims to provide an anti-aging traditional Chinese medicine extract and an extraction method thereof, and a product with better anti-aging effect can be obtained.

In order to solve the problems, the invention provides a composition which comprises the following components in parts by weight: 20-30 parts of astragaloside IV, 5-5 parts of ginsenoside Rg 11, 2-6 parts of ginsenoside Re and 10.2-0.5 part of ginsenoside Rb.

Further, the composition comprises the following components in parts by weight: 22-28 parts of astragaloside IV, 11.8-4 parts of ginsenoside Rg, 3-5 parts of ginsenoside Re and 10.2-0.3 part of ginsenoside Rb.

The invention also provides an extract which comprises the following raw materials in parts by weight: 1 part of ginseng and/or American ginseng, 4-8 parts of astragalus, and further 4-6 parts of astragalus; the extract contains 20-30% of astragaloside IV, 20-5% of ginsenoside Rg 11, 2-6% of ginsenoside Re and 10.1-0.5% of ginsenoside Rb.

Furthermore, the extract contains 22-28% of astragaloside IV, 11.8-4% of ginsenoside Rg, 3-5% of ginsenoside Re and 10.2-0.3% of ginsenoside Rb.

Further, the total saponin content in the extract is 75-95%, and the total polysaccharide content is 3-10%.

Furthermore, the content of the total saponins is 80-90%, and the content of the total polysaccharides is 5-8%.

Further, the ginseng is ginseng and/or a ginseng processed product.

Further, the processed ginseng product is selected from one or more of sun-dried ginseng, red ginseng and active ginseng.

The invention also provides an extract comprising ginsenoside Re; the high performance liquid chromatography fingerprint spectrum of the extract at least comprises 6 characteristic peaks, the characteristic peak of the ginsenoside Re is taken as a reference peak, and the relative retention time of the 6 characteristic peaks and the reference peak is as follows: 0.95, 1.00, 1.80, 2.13, 2.36 and 3.20.

Further, the method also comprises 11 characteristic peaks, and the relative retention time of the 11 characteristic peaks and a reference peak is as follows: 1.66, 1.69, 1.73, 1.86, 1.88, 1.94, 1.99, 2.02, 2.28, 2.40, 2.45.

Further, the raw materials comprise the following raw materials in parts by weight: 1 part of ginseng and/or American ginseng, 4-8 parts of astragalus, and further 4-6 parts of astragalus.

Further, the ginseng is ginseng and/or a ginseng processed product.

Further, the processed ginseng product is selected from one or more of sun-dried ginseng, red ginseng and active ginseng.

Further, the relative peak areas of the characteristic peaks with relative retention times of 0.95, 1.00, 1.80, 2.13, 2.36 and 3.20 are, in order: 48.41%, 100.00%, 107.04%, 48.70%, 340.51% and 54.83%.

Further, the relative peak areas of the characteristic peaks with relative retention times of 1.66, 1.69, 1.73, 1.86, 1.88, 1.94, 1.99, 2.02, 2.28, 2.40 and 2.45 are, in order: 9.52%, 17.21%, 2.61%, 4.16%, 22.88%, 10.92%, 16.34%, 8.33%, 3.37%, 5.14% and 14.59%.

The relative retention time, relative peak area, and the like may all vary within reasonable limits, such as within 5% of the value, and are considered to fall within the scope of the claimed invention.

The invention also provides a detection method of the high performance liquid chromatography fingerprint spectrum of the extract, which adopts high performance liquid chromatography to carry out qualitative and/or quantitative detection, and the chromatographic conditions comprise:

a chromatographic column: c18 silica gel column;

mobile phase: water and acetonitrile, using the following gradient elution procedure:

time (min)	Water (V%)	Acetonitrile (V%)
			0	80～90	10～20
30～35	60～70	30～40
			70～85	10～30	70～90

；

Further, the following elution procedure was employed:

time (min)	Water (V%)	Acetonitrile (V%)
			0	85	16
33	66	34
			78.5	20	80

。

Further, the detection method further comprises the following conditions:

specification of chromatographic column: 150 x 3.9mm, 5 μm;

column temperature: 25-35 ℃, preferably 30 ℃;

flow velocity: 0.8-1.2 mL/min, preferably 1.00 mL/min;

detecting wavelength: 200-205 nm, preferably 203 nm.

The invention also provides a preparation method of the natural plant extract, which comprises the following steps: (1) extracting the raw materials with water; (2) enriching by adopting macroporous adsorption resin; (3) carrying out enzymolysis; (4) performing membrane filtration, wherein the molecular weight cutoff is 200-1000; (5) solid-liquid separation;

the plant is selected from one or two of ginseng and American ginseng, and/or astragalus; that is, the plant may be one of Ginseng radix, radix Panacis Quinquefolii, and radix astragali; it can also be: mixing and extracting ginseng and astragalus, American ginseng and astragalus, and ginseng and American ginseng; or extracting Ginseng radix, radix Panacis Quinquefolii, and radix astragali.

Further, the ginseng is ginseng and/or a ginseng processed product.

Further, the processed ginseng product is selected from one or more of sun-dried ginseng, red ginseng and active ginseng.

The sun-dried ginseng is a commercial ginseng, also called white ginseng, which is a product obtained by digging out roots of ginseng, cleaning and drying in the sun.

Ginseng radix Rubri is prepared by steaming Ginseng radix of Araliaceae, collecting and digging in autumn, cleaning, steaming, and drying. The present invention uses general red ginseng.

The active ginseng is low-temperature freeze-dried ginseng. The product can quickly absorb water when placed in water, and the appearance can be restored to the state similar to that of fresh ginseng, so the product is called as 'active ginseng'.

The sun-dried ginseng, the red ginseng and the active ginseng are prepared by common ginseng processed products in the field, can be purchased in the market and can be prepared by the prior art, and the obtained sun-dried ginseng, the red ginseng and the active ginseng can be applied to the invention, and are preferably the sun-dried ginseng, the red ginseng and the active ginseng which meet the national standard (the national standard document: the quality of the sun-dried ginseng, the red ginseng and the active ginseng, the GB/T22536 and 2018, the quality of the red ginseng and the GB/T22538 and 2018, the quality of the active ginseng and the like).

In an embodiment of the present invention, in the water extraction step, the water: the liquid-material ratio of the raw materials is 5-15: 1 mL/g;

further, the number of times of water extraction is 1-4; further, it is 2 times.

In the embodiment of the invention, the macroporous adsorption resin is used for enriching, an extracting solution obtained by water extraction is adsorbed by using macroporous adsorption resin, then is washed by water, 10-40% ethanol and 60-80% ethanol in sequence, and a part obtained by washing 60-80% ethanol is collected;

further, washing with water, 25-30% ethanol and 70-80% ethanol respectively, and collecting the parts obtained by washing with 70-80% ethanol;

further, the macroporous adsorption resin is nonpolar or weak polar macroporous adsorption resin; preferably, the macroporous adsorption resin is selected from D-101, HPD-10, X-5 and AB-8.

In the embodiment of the invention, the enzymolysis is to perform enzymolysis on the substance obtained after the macroporous adsorption resin is enriched;

the enzymolysis can be carried out by one or more of glycosidase, pectinase and amylase.

Further, the enzymolysis may be performed by using one or more of β -glucosidase, xylosidase, lactase, arabinofuranosidase, pectin lyase, and amylase.

In a specific embodiment of the invention, the enzyme is a complex enzyme consisting of beta-glucosidase and xylosidase in a weight ratio of 2-4: 1.

Further, the enzymolysis conditions are as follows: performing enzymolysis on the raw materials at the conditions of a complex enzyme with the mass of 0.2-1.0% and the pH of 5-6 and the temperature of 40-55 ℃;

further, the using amount of the complex enzyme is 0.4-0.6%;

further, the enzymolysis time is 3-8 h; furthermore, the enzymolysis time is 4-6 h.

The invention also provides one of the following extracts prepared by the method:

(1) a ginseng extract; (2) american ginseng extract; (3) an extract of Astragalus membranaceus; (4) a mixed extract of ginseng and American ginseng; (5) a mixed extract of ginseng and astragalus; (6) a mixed extract of American ginseng and astragalus; (7) mixed extracts of Ginseng radix, radix Panacis Quinquefolii and radix astragali.

The invention also provides a composition, which comprises the following raw materials in parts by weight: 1 part of the ginseng extract and/or the American ginseng extract and 5-14 parts of the astragalus extract; further, the traditional Chinese medicine composition comprises 1 part of the ginseng extract and/or the American ginseng extract and 5.5-11 parts of the astragalus extract.

The invention also provides the application of one or more of the extract, the extraction method and the composition in preparing products capable of improving telomerase activity and/or increasing telomere length; the extract described herein may be selected from all of the extracts mentioned above, not limited to extracts defined by composition or fingerprint.

Further, the product is telomerase activator;

further, the product is an anti-aging product;

further, the product is a product for delaying cell aging.

The invention also provides a product, which comprises one or more of the extract and the composition; the extract described herein may be selected from all of the extracts mentioned above, not limited to extracts defined by composition or fingerprint.

Products mentioned in the present invention include, but are not limited to, food, pharmaceutical, health care products.

The product can also comprise adjuvants and auxiliary components in addition to the above extract or composition.

The pharmaceutically acceptable auxiliary materials are general names of all the additional materials except the main medicine in the medicine, and the auxiliary materials have the following properties: (1) no toxic effect on human body and few side effects; (2) the chemical property is stable and is not easily influenced by temperature, pH, storage time and the like; (3) has no incompatibility with the main drug, and does not influence the curative effect and quality inspection of the main drug; (4) does not interact with the packaging material. The auxiliary materials in the invention include, but are not limited to, a filler (diluent), a lubricant (glidant or anti-adhesion agent), a dispersing agent, a wetting agent, an adhesive, a regulator, a solubilizer, an antioxidant, a bacteriostatic agent, an emulsifier, a disintegrating agent and the like. The binder comprises syrup, acacia, gelatin, sorbitol, tragacanth, cellulose and its derivatives (such as microcrystalline cellulose, sodium carboxymethylcellulose, ethyl cellulose or hydroxypropyl methylcellulose), gelatin slurry, syrup, starch slurry or polyvinylpyrrolidone; the filler comprises lactose, sugar powder, dextrin, starch and its derivatives, cellulose and its derivatives, inorganic calcium salt (such as calcium sulfate, calcium phosphate, calcium hydrogen phosphate, precipitated calcium carbonate, etc.), sorbitol or glycine, etc.; the lubricant comprises superfine silica gel powder, magnesium stearate, talcum powder, aluminum hydroxide, boric acid, hydrogenated vegetable oil, polyethylene glycol and the like; the disintegrating agent comprises starch and its derivatives (such as sodium carboxymethyl starch, sodium starch glycolate, pregelatinized starch, modified starch, hydroxypropyl starch, corn starch, etc.), polyvinylpyrrolidone or microcrystalline cellulose, etc.; the wetting agent comprises sodium lauryl sulfate, water or alcohol, etc.; the antioxidant comprises sodium sulfite, sodium bisulfite, sodium pyrosulfite, dibutylbenzoic acid, etc.; the bacteriostatic agent comprises 0.5% of phenol, 0.3% of cresol, 0.5% of chlorobutanol and the like; the regulator comprises hydrochloric acid, citric acid, potassium (sodium) hydroxide, sodium citrate, and buffer (including sodium dihydrogen phosphate and disodium hydrogen phosphate); the emulsifier comprises polysorbate-80, sorbitan fatty acid, pluronic F-68, lecithin, soybean lecithin, etc.; the solubilizer comprises Tween-80, bile, glycerol, etc.

The auxiliary components can be traditional Chinese medicinal materials with the same function or function as the extract or the composition of the invention or substances such as extracts, chemicals and the like of the traditional Chinese medicinal materials, and can also be active substances with other functions or efficacies.

The invention has the beneficial effects that:

(1) the ginseng extract, the astragalus extract and the extract obtained by mixing and extracting the ginseng and the astragalus can effectively improve the activity of telomerase, have high anti-aging activity and can be used for preparing anti-aging products.

(2) The anti-aging activity of the ginseng and astragalus extract is better than that of any one extract under the same concentration, which shows that the ginseng and astragalus extract of the invention plays a synergistic effect after being combined, so that the anti-aging performance of the ginseng and astragalus extract is obviously improved.

Drawings

FIG. 1 shows the fingerprint of the extract of ginseng and astragalus membranaceus of example 1 of the present invention.

Detailed Description

The present invention is further illustrated by the following specific examples and specific tests.