Ginseng and ganoderma lucidum preparation with anti-oxidation and anti-aging effects and application of ginseng and ganoderma lucidum preparation

Abstract

The invention relates to a ginseng and ganoderma lucidum preparation with anti-oxidation and anti-aging effects and application of the ginseng and ganoderma lucidum preparation with the anti-oxidation and anti-aging effects, ginseng and ganoderma lucidum are used as main components, and the ginseng and ganoderma lucidum preparation with the anti-oxidation and anti-aging effects is obtained through a specific preparation technology.

Description

Technical Field

[0001] The invention relates to the field of health food, and in particular to a ginseng and ganoderma lucidum preparation with anti-oxidation and anti-aging effects and an application thereof. Background Art

[0002] With the deepening of modern scientific research, the active ingredients and mechanisms of action of the two medicinal materials, Panax ginseng and Ganoderma lucidum, have been gradually revealed. Both have rich pharmacological activities and health functions. Ginseng is well-known for its effects of enhancing immunity, anti-fatigue, and anti-aging, while Ganoderma lucidum is valued for its multiple biological activities such as immunomodulation, anti-tumor, and anti-oxidation, and has been widely used in the field of medicine and health care.

[0003] Ginseng contains a variety of active ingredients, such as ginsenosides, polysaccharides, volatile oils, amino acids, etc. Ginsenosides are the main active ingredients in ginseng, which have the effects of enhancing immunity, resisting fatigue, and improving cardiovascular and cerebrovascular functions. Ginseng polysaccharides also have the effects of enhancing immunity, anti-tumor, and anti-oxidation.

[0004] Ganoderma lucidum polysaccharides (GLPs) are one of the earliest and most intensively studied active ingredients in Ganoderma lucidum. Ganoderma lucidum polysaccharides have biological activities such as immunomodulation, anti-tumor, antioxidant, and anti-aging. Triterpenoid compounds in Ganoderma lucidum also have significant pharmacological activities.

[0005] The combination of ginseng and ganoderma can achieve the synergistic effect of the two medicinal materials. The ginseng and ganoderma combination has shown significant effects in improving the body's immunity, anti-fatigue, anti-aging, etc. In addition, the ginseng and ganoderma combination also shows potential application prospects in anti-tumor, improving cardiovascular and cerebrovascular function, regulating blood sugar, etc.

[0006] However, traditional extraction methods such as hot water extraction and ultrasonic-assisted extraction often have problems such as low extraction efficiency and loss of active ingredients. In addition, the active ingredients in ginseng and ganoderma are complex, and traditional extraction methods are difficult to achieve efficient separation and purification of active ingredients.

[0007] Ganoderma lucidum is a medicinal fungus with rich pharmacological activities. Its polysaccharide components (GLPs) have attracted widespread attention due to their multiple biological activities such as immunomodulation, anti-tumor, and antioxidant.

[0008] The existing Ganoderma lucidum polysaccharide extraction technologies have their own advantages and disadvantages. Hot water extraction is low-cost but inefficient; ultrasonic and microwave-assisted extraction can improve efficiency but may affect heat-sensitive components; supercritical fluid extraction is efficient but expensive.

[0009] High-voltage pulse electric field extraction technology is a new type of non-thermal processing technology. Through instantaneous high-voltage electric field treatment, the cell membrane undergoes electroporation, thereby increasing the diffusion of intracellular substances and improving the extraction efficiency. This technology has the advantages of high extraction efficiency, simple operation, and little damage to active ingredients.

[0010] The Sevage method is a classic protein removal method. It effectively removes proteins from aqueous solutions and improves the purity of polysaccharides through mixed solvent treatment. Summary of the invention

[0011] In view of the shortcomings of the prior art, the present invention provides a method for extracting polysaccharides from Ganoderma lucidum by using a high-voltage pulsed electric field combined with the Sevage method. The method can improve the extraction efficiency and purity while maintaining the biological activity of Ganoderma lucidum polysaccharides, providing technical support for the further research and application of ginseng and Ganoderma lucidum compositions.

[0012] One of the technical problems to be solved by the present invention is: to provide a method for extracting Ganoderma lucidum polysaccharides by using a high-voltage pulsed electric field combined with a Sevage method. The high-voltage pulsed electric field can increase the permeability of the cell membrane in a short period of time, which is conducive to the release of polysaccharides; the Sevage method can effectively remove proteins and improve the purity of polysaccharides.

[0013] The second technical problem to be solved by the present invention is to provide a ginseng and ganoderma lucidum preparation with antioxidant and anti-aging effects. The ginseng and ganoderma lucidum oral liquid obtained by the method can effectively remove free radicals and show excellent antioxidant and anti-aging effects.

[0014] The beneficial effects of the present invention are as follows: 1. The method can improve the extraction efficiency and purity while maintaining the biological activity of ganoderma lucidum polysaccharide, has the advantages of short time, low cost, simple operation, etc., and is suitable for industrial production. 2. The prepared ginseng ganoderma oral liquid preparation has excellent antioxidant and anti-aging effects.

[0015] The extraction method of the ganoderma lucidum polysaccharide extract of the present invention is as follows: S1. High-pressure pulse method for extracting Ganoderma lucidum: (1) Grind the fruiting bodies of Ganoderma lucidum and pass them through a 20-200 mesh sieve to obtain Ganoderma lucidum powder. Weigh an appropriate amount of Ganoderma lucidum powder, add 10-20 times the volume of deionized water, and swell at 20-40° C. for 30 minutes; (2) treating the swollen mixture in a high-voltage pulse electric field under the following conditions: electric field strength of 20-60 kV / cm, frequency of 100-1000 Hz, number of pulses of 10-60, treatment time of 10-30 min, centrifuging the treated mixture and collecting the supernatant; S2. Sevage method to remove protein: Add an appropriate amount of Sevage reagent to the collected supernatant, mix according to a volume ratio of 1:2-1:10, stand at 4-20°C for 0.5-2 hours, take the supernatant after centrifugation, repeat the operation 3-6 times until the protein is removed, use a rotary evaporator to concentrate the supernatant to 1 / 5 of the original volume, add 95% ethanol to precipitate polysaccharides, and stand at 4°C overnight; The precipitate is collected, washed with anhydrous ethanol and acetone, and dried in vacuum. The dried polysaccharide is dissolved in deionized water, dialyzed to remove small molecular impurities, and the dialyzed polysaccharide solution is freeze-dried to obtain a refined Ganoderma lucidum polysaccharide extract.

[0016] Furthermore, the mesh size of the sieve in the above step (1) is 60 mesh, and 10 times the volume of deionized water is added, and the mixture is swelled at 30° C. for 30 minutes.

[0017] Furthermore, in the above step (2), the electric field strength is 40 kV / cm, the frequency is 500 Hz, the number of pulses is 40, and the processing time is 20 min.

[0018] Furthermore, in step S2, the mixture is mixed in a volume ratio of 1:3 (i.e., the volume ratio of Sevage reagent to supernatant is 1:3), and allowed to stand at 4° C. for 1 hour. In step S2, the volume ratio of chloroform to n-butanol in the Sevage reagent is 4:1.

[0019] Further, step S2 is repeated 4 times until no large amount of white precipitate is generated in the middle layer.

[0020] The preparation method of the ginseng extract of the present invention is as follows: take ginseng, cut it into thick slices, add water and boil it twice, the first time for 2 hours and the second time for 1.5 hours, filter the decoction, combine the filtrate, pass it through a D101 macroporous adsorption resin column, elute with water until it is colorless, then elute with 60% ethanol, collect the 60% ethanol eluate, concentrate the eluate to a clear paste with a relative density of 1.06 to 1.08 (80°C), dry it, and crush it to obtain the ginseng extract.

[0021] The preparation method of the preparation of the invention is as follows: weigh 5-10 parts of ginseng extract and 10-30 parts of ganoderma lucidum polysaccharide extract, add 100-300 parts of water, stir and mix evenly, filter to remove impurities, fill, and sterilize at high temperature to obtain ginseng and ganoderma lucidum oral liquid.

[0022] Furthermore, 8 parts of ginseng extract and 20 parts of ganoderma lucidum extract are added with 200 parts of water, stirred and mixed evenly, filtered to remove impurities, filled, and sterilized at high temperature to obtain the ginseng and ganoderma lucidum oral liquid.

[0023] The present invention adopts phenol-sulfuric acid method to detect polysaccharide content and adopts free radical scavenging experiment to determine the antioxidant effect of ginseng and ganoderma oral liquid.

[0024] The invention uses ganoderma lucidum as raw material, and extracts the main component ganoderma lucidum polysaccharide through high-voltage pulse electric field combined with Sevage method. The method is simple in process, can improve the yield of ganoderma lucidum polysaccharide, has the advantages of short time and good effect, can also maintain its physiological activity to the maximum extent, has high polysaccharide extraction rate and low cost, and meets the needs of large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 These are the polysaccharide extraction results of each group.

[0026] Figure 2 is the DPPH free radical scavenging rate of each group. DETAILED DESCRIPTION

[0027] The present invention is further described below by specific examples and comparative examples. However, it should be understood that these examples and comparative examples are only used for more detailed and specific description and should not be construed as limiting the present invention in any form.

[0028] Example 1

[0029] The ganoderma lucidum fruiting bodies were crushed and passed through a 60-mesh sieve to obtain ganoderma lucidum powder. 200 g of the ganoderma lucidum powder was weighed and defatted with ether using a Soxhlet extractor to remove fat-soluble impurities and color.

[0030] The defatted Ganoderma lucidum residue was dried and extracted with boiling water to obtain a Ganoderma lucidum water extract. The extract was then concentrated under reduced pressure at 60°C and centrifuged to obtain a supernatant. Anhydrous ethanol was added to the supernatant for precipitation. After standing at 4°C overnight, the precipitated polysaccharides were collected by centrifugation and the polysaccharide content was detected by the phenol-sulfuric acid method. The final Ganoderma lucidum polysaccharide yield was 1.2% and the purity was 60.7%.

[0031] Take ginseng, cut it into thick slices, add water and boil it twice, the first time for 2 hours, the second time for 1.5 hours, filter the decoction, combine the filtrate, pass it through a D101 macroporous adsorption resin column, elute with water until it is colorless, then elute with 60% ethanol, collect the 60% ethanol eluate. The filtrate is concentrated to a clear paste with a relative density of 1.06-1.08 (80°C), dried, and crushed to obtain ginseng extract.

[0032] Weigh 8 parts of ginseng extract and 20 parts of ganoderma lucidum extract respectively, add 200 parts of water, stir and mix evenly, filter to remove impurities, fill, and sterilize at high temperature to prepare the ginseng and ganoderma lucidum oral liquid.

[0033] Example 2

[0034] The difference between this embodiment and embodiment 1 is that the extraction and purification methods of Ganoderma lucidum polysaccharide are different.

[0035] S1. High-pressure pulse method for extracting Ganoderma lucidum: (1) Ganoderma lucidum fruiting bodies were crushed and passed through a 60-mesh sieve to obtain Ganoderma lucidum powder. 200 g of Ganoderma lucidum powder was weighed, 10 times the volume of deionized water was added, and the mixture was swollen at 30° C. for 30 minutes; (2) The swollen mixture is treated in a high-voltage pulse electric field, and the electric field strength, frequency, number of pulses, and treatment time are adjusted. The adjustment parameters are shown in Table 1. The treated mixture is centrifuged and the supernatant is collected.

[0036] The electric field intensity, frequency, number of pulses, and processing time of the high-voltage pulse extraction parameters were adjusted to evaluate the yield of Ganoderma lucidum polysaccharides obtained with different parameters. The experiment was repeated three times and the average value was taken. The polysaccharide extraction results are shown in Figure 1 .

[0037] Table 1: Groups and parameters of Ganoderma lucidum extraction by high pressure pulse method

[0038] The results are as follows Figure 1 As shown, Example 1 adopts a conventional method for extracting ganoderma lucidum polysaccharide, and the polysaccharide yield is 1.2% and the purity is 60.7%. This is also the current general extraction level of ganoderma lucidum polysaccharide. When using the high-voltage pulse method to extract ganoderma lucidum polysaccharide, it is not as expected that any parameters can reach the above-mentioned extraction level. For example, the extraction rates of groups such as 1-1, 1-2, 1-3, and 1-4 are all lower than the above-mentioned conventional extraction method. Parameters such as electric field strength and frequency are too low, resulting in insufficient extraction and low polysaccharide extraction yield. If the processing time is too long, it is easy to damage ganoderma lucidum polysaccharide, resulting in a decrease in yield. Wherein the electric field strength is 40kV / cm, the frequency is 500Hz, the number of pulses is 40, and the polysaccharide yield is the highest when the processing time is 20min.

[0039] S2. Sevage method to remove protein: Select the parameters of groups 1-8 in step S1 to perform the Sevage method for protein removal.

[0040] Add an appropriate amount of Sevage reagent to the collected supernatant, mix according to a certain volume ratio, stand at 4°C, take the supernatant after centrifugation, and repeat the operation 3-6 times (adjust according to experimental needs, 4 times is sufficient for most groups) until the protein is removed. Use a rotary evaporator to concentrate the supernatant to 1 / 5 of the original volume, add 95% ethanol to precipitate polysaccharides, and stand at 4°C overnight.

[0041] The precipitate is collected, washed with anhydrous ethanol and acetone, and dried in vacuum. The dried polysaccharide is dissolved in deionized water and dialyzed to remove small molecular impurities. The dialyzed polysaccharide solution is freeze-dried to obtain refined Ganoderma lucidum polysaccharide.

[0042] The volume ratio of chloroform to n-butanol, the volume ratio of Sevage reagent to supernatant (polysaccharide solution), and the standing time involved in the above operation process were adjusted. The adjustment parameters are shown in Table 2. The polysaccharide content was detected by phenol-sulfuric acid method, and the purity of polysaccharides was detected by gel chromatography. The total polysaccharide yield and the purity of polysaccharides after steps S1 and S2 were evaluated. The experiment was repeated three times and the average value was taken.

[0043] Table 2: Sevage protein removal groups and parameters

[0044] The results are shown in Table 2. After deproteinization by the Sevage method, the total yield of polysaccharides is somewhat lost compared to the polysaccharides extracted from Ganoderma lucidum by the high-pressure pulse method in step S1. This may be due to the partial polysaccharide encapsulated in the protein, but the overall loss is not large. However, the purity of Ganoderma lucidum polysaccharides has been significantly improved, and its purity is much higher than the 60.7% polysaccharide purity in Example 1. The above-mentioned groups 2-4, that is, the Sevage reagent is configured, the solution is configured according to the volume ratio of chloroform and n-butanol = 4:1, and the volume ratio of Sevage reagent: supernatant (polysaccharide solution) = 1:3, and it is left to stand for 1 hour. The purity and total polysaccharide yield of the Ganoderma lucidum polysaccharide obtained reach the maximum value.

[0045] Example 3

[0046] Prepare the refined Ganoderma lucidum polysaccharide extracts obtained from Groups 2-1, 2-2, 2-3, 2-4, 2-5, and 2-6 in Example 2; Prepare ginseng extract as follows: Take ginseng, cut into thick slices, add water and boil twice, the first time for 2 hours, the second time for 1.5 hours, filter the decoction, combine the filtrate, pass it through a D101 macroporous adsorption resin column, elute with water until colorless, then elute with 60% ethanol, collect the 60% ethanol eluate. Concentrate the filtrate to a clear paste with a relative density of 1.06 to 1.08 (80°C), dry it, and crush it to obtain ginseng extract.

[0047] The preparation method of the preparation of the present invention is as follows: weigh 8 parts of ginseng extract and 20 parts of ganoderma lucidum polysaccharide extract, add 200 parts of water, stir and mix evenly, filter to remove impurities, fill, and sterilize at high temperature to obtain ginseng and ganoderma oral liquid 2-1, 2-2, 2-3, 2-4, 2-5, and 2-6.

[0048] Effect experiment (antioxidant activity determination) DPPH free radical scavenging experiment Take 2 mL of the test solution and 2 mL of DPPH ethanol solution, mix them evenly, react in the dark at room temperature (30 min), centrifuge at 8 000 rpm for 10 min at room temperature, and measure the absorbance of the supernatant at 517 nm. The measurement was repeated for three groups in parallel.

[0049] Experimental method: Take 2mL of ginseng and ganoderma oral liquid and add it to 2mL of 0.1mmol / L freshly prepared DPPH-anhydrous ethanol solution, mix well, react at room temperature in the dark for 30min, centrifuge at 10000r / min for 2min, take the supernatant and measure the absorbance at 517nm wavelength, record it as Ai. For the blank group, replace the DPPH solution with an equal volume of anhydrous ethanol and measure the absorbance, record it as Aj; DPPH free radical scavenging rate (%) = [1-(Ai-Aj) / Ac] × 100%, where Ai is the absorbance of the sample group; Ac is the absorbance of the control group (distilled water replaces the sample); Aj is the absorbance of the blank group (anhydrous ethanol replaces DPPH). Figure 2 .

[0050] It can be seen that the ginseng and ganoderma oral liquid of the present invention has an obvious effect on scavenging free radicals, and the ginseng and ganoderma oral liquid obtained by the specific preparation method of the present invention has a significantly better free radical scavenging effect than the ginseng and ganoderma oral liquid prepared by the conventional method.

[0051] Without departing from the scope of the present invention, those of ordinary skill in the art may implement modifications and variations to the present invention. In addition, it should be understood that the aspects of the various embodiments may be interchangeable in whole or in part. The foregoing description is exemplary only and is not intended to limit the present invention further described in the appended claims.

Claims

1. A Ganoderma lucidum and ginseng preparation, wherein: Contains Ganoderma lucidum polysaccharide extract and Ginseng extract.

2. The preparation according to claim 1, wherein This preparation is in the form of an oral solution.

3. The preparation according to claim 1 or 2, wherein The preparation method of the preparation is as follows: weigh 5-10 parts of ginseng extract and 10-30 parts of ganoderma lucidum polysaccharide extract, add 100-300 parts of water, stir and mix evenly, filter to remove impurities, fill, and sterilize at high temperature to obtain ginseng and ganoderma lucidum oral liquid.

4. The preparation according to any one of claims 1 to 3, wherein The extraction steps of Ganoderma lucidum polysaccharide extract are as follows: S1. High-pressure pulse method for extracting Ganoderma lucidum: (1) Grind the fruiting bodies of Ganoderma lucidum and pass them through a 20-200 mesh sieve to obtain Ganoderma lucidum powder. Weigh an appropriate amount of Ganoderma lucidum powder, add 10-20 times the volume of deionized water, and swell at 20-40° C. for 30 minutes; (2) treating the swollen mixture in a high-voltage pulse electric field under the following conditions: electric field strength of 20-60 kV / cm, frequency of 100-1000 Hz, number of pulses of 10-60, treatment time of 10-30 min, centrifuging the treated mixture and collecting the supernatant; S2. Sevage method to remove protein: Add an appropriate amount of Sevage reagent to the collected supernatant, mix according to a volume ratio of 1:2-1:10, stand at 4-20°C for 0.5-2 hours, take the supernatant after centrifugation, repeat the operation until the protein is removed, use a rotary evaporator to concentrate the supernatant to 1 / 5 of the original volume, add 95% ethanol to precipitate polysaccharides, and stand at 4°C overnight; The precipitate is collected, washed with anhydrous ethanol and acetone, and vacuum dried. The dried polysaccharide is dissolved in deionized water and dialyzed to remove small molecular impurities. The dialyzed polysaccharide solution is freeze-dried to obtain a refined Ganoderma lucidum polysaccharide extract.

5. The preparation according to claim 4, wherein The mesh size of the sieve in step (1) is 60 mesh, 10 times the volume of deionized water is added, and the mixture is swelled at 30° C. for 30 minutes.

6. The preparation according to claim 4 or 5, wherein In the step (2), the electric field strength is 40 kV / cm, the frequency is 500 Hz, the number of pulses is 40, and the processing time is 20 min.

7. The preparation according to any one of claims 4 to 6, wherein In step S2, the Sevage method is used to remove protein, the volume ratio of chloroform to n-butanol in the Sevage reagent is 4:1, the volume ratio of the Sevage reagent to the supernatant is 1:3, and the standing time is 1 hour.

8. The preparation according to any one of claims 1 to 7, wherein The preparation method of ginseng extract is as follows: take ginseng, cut it into thick slices, add water and boil it twice, the first time for 2 hours and the second time for 1.5 hours, filter the decoction, combine the filtrate, pass it through a D101 macroporous adsorption resin column, elute with water until colorless, then elute with 60% ethanol, collect the 60% ethanol eluate, concentrate the eluate to a clear paste with a relative density of 1.06 to 1.08 (80°C), dry it, and crush it to obtain the ginseng extract.

9. Use of the preparation according to any one of claims 1 to 8 in the preparation of antioxidant and anti-aging health products, health foods or health medicines.

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