Preparation method and application of composite freeze-dried powder based on inonotus obliquus

Abstract

The invention discloses a preparation method and application of composite freeze-dried powder based on inonotus obliquus, and relates to the technical field of biological product preparation. The invention aims to solve the problems that the extraction efficiency and the extraction rate of effective components in inonotus obliquus are low, the stability and the activity of the effective components are difficult to maintain, and the provision condition is harsh due to low solubility of schisandra chinensis. According to the inonotus obliquus composite freeze-dried powder prepared by compounding the fructus schisandrae chinensis extract and the amomum tsao-ko alcohol extract by adopting the process disclosed by the invention, the content of crude polysaccharide is increased by 27.67%, the content of inonotus obliquus polysaccharide is increased by 23.2%, the content of beta-glucan is increased by 14.28%, the content of triterpenoids is increased by 26.2%, the content of schizandrin is increased by 40%, and the content of fructus schizandrin is increased by 20%. The compound has the potential of being used for preparing drugs with high curative effects, such as tumor resistance, blood sugar reduction, blood pressure reduction, virus resistance and aging resistance. The invention can obtain the preparation method and the application of the inonotus obliquus-based composite freeze-dried powder.

Description

Technical Field

[0001] The present invention relates to the technical field of biological product preparation, and in particular to a preparation method and application of Inonotus obliquus composite freeze-dried powder. Background Art

[0002] Inonotus obliquus, also known as Chaga, belongs to the family of Inonotaceae and is mainly distributed in temperate regions. Inonotus obliquus is a medicinal fungus that grows on birch trees. Its sclerotium is tumor-shaped, black-gray on the outside, yellow on the inside, and thin, dark brown in color. Inonotus obliquus contains various chemical components such as betulin, betulinol, oxidized triterpenoids, lanosterol-type triterpenoids, folic acid derivatives, aromatic vanillic acid, syringic acid and gamma-hydroxybenzoic acid.

[0003] The various active ingredients in Inonotus birch have various biological activities such as anti-cancer, hypoglycemic, hypotensive, antiviral, and anti-aging. Therefore, in recent years, the extraction process of Inonotus birch has been increasingly developed. At present, the extraction process of Inonotus birch usually takes a long time, resulting in the loss of active ingredients; at the same time, the extraction efficiency of various active ingredients in Inonotus birch is low, and the biological activity is difficult to maintain after extraction.

[0004] Schisandra chinensis belongs to the Schisandrae family and is divided into northern Schisandra and southern Schisandra. It mainly contains lignans, schisandrin, schisandrin, vitamin C, resins and tannins, etc. It has the effects of astringing the lungs and relieving cough, nourishing and astringing essence, and stopping diarrhea and sweating. Among them, the solubility of schisandrin in the ingredients is very low, and its extraction process is limited by many factors such as temperature, solvent properties and pressure. Summary of the invention

[0005] The purpose of the present invention is to solve the problems that the extraction efficiency and extraction rate of the effective ingredients in Inonotus obliquus are low, the stability and activity of the effective ingredients are difficult to maintain, and the provision conditions are harsh due to the low solubility of Schisandra chinensis, and to provide a preparation method and application of Inonotus obliquus composite freeze-dried powder.

[0006] A preparation method based on Inonotus obliquus composite freeze-dried powder is carried out according to the following steps:

[0007] Step S1: Extraction of Inonotus obliquus;

[0008] The pretreated Inonotus birchii is mixed with water, and then placed in a microwave reactor, heated to 90-110° C., and kept warm at 90-110° C. for 15-20 minutes to obtain an Inonotus birchii water extract; the Inonotus birchii water extract is sterilized and filtered using a microporous filter membrane to obtain a sterile Inonotus birchii water extract;

[0009] Step S2: extracting Schisandra chinensis;

[0010] The schisandra chinensis is mixed with a polar solvent, heated to 45-55° C., and extracted for 5-10 minutes under pressure, ultrasound assistance and high-voltage pulse electric field conditions, purified after the extraction, and then filtered using a microporous filter membrane to obtain a schisandra chinensis extract;

[0011] Step S3: preparation of composite freeze-dried powder;

[0012] The sterile Inonotus obliquus water extract obtained in step S1, the Schisandra chinensis extract obtained in step S2 and the Amomum villosum alcohol extract are fully mixed, and then placed in a freeze dryer for freeze drying to obtain a composite freeze-dried powder based on Inonotus obliquus;

[0013] The volume ratio of the sterile inonotus birchii water extract, the northern schisandra chinensis extract and the tsaoko alcohol extract is (8-10): (3-5): (1-3).

[0014] An application of a composite freeze-dried powder based on Inonotus obliquus, and the application of the composite freeze-dried powder based on Inonotus obliquus in the preparation of anti-tumor, hypoglycemic, hypotensive, antiviral and anti-aging drugs.

[0015] Beneficial effects of the present invention:

[0016] The composite freeze-dried powder of Inonotus obliquus prepared by the process of the present invention and compounding with Schisandra chinensis extract and Amomum villosum alcohol extract has a crude polysaccharide content increased by 27.67%, a polysaccharide content increased by 23.2%, a beta-glucan content increased by 14.28%, a triterpenoid compound content increased by 26.2%, and a schisandra alcohol A content increased by 40% compared with similar freeze-dried powder of Inonotus obliquus on the market, and has the potential to be used for preparing highly effective anti-tumor, hypoglycemic, hypotensive, antiviral and anti-aging drugs.

[0017] The present invention can obtain a preparation method and application of Inonotus obliquus composite freeze-dried powder. DETAILED DESCRIPTION

[0018] Specific implementation method 1: This implementation method is a preparation method based on Inonotus obliquus composite freeze-dried powder, which is carried out according to the following steps:

[0019] Step S1: Extraction of Inonotus obliquus;

[0020] The pretreated Inonotus birchii is mixed with water, and then placed in a microwave reactor, heated to 90-110° C., and kept warm at 90-110° C. for 15-20 minutes to obtain an Inonotus birchii water extract; the Inonotus birchii water extract is sterilized and filtered using a microporous filter membrane to obtain a sterile Inonotus birchii water extract;

[0021] Step S2: extracting Schisandra chinensis;

[0022] The schisandra chinensis is mixed with a polar solvent, heated to 45-55° C., and extracted for 5-10 minutes under pressure, ultrasound assistance and high-voltage pulse electric field conditions, purified after the extraction, and then filtered using a microporous filter membrane to obtain a schisandra chinensis extract;

[0023] Step S3: preparation of composite freeze-dried powder;

[0024] The sterile Inonotus obliquus water extract obtained in step S1, the Schisandra chinensis extract obtained in step S2 and the Amomum villosum alcohol extract are fully mixed, and then placed in a freeze dryer for freeze drying to obtain a composite freeze-dried powder based on Inonotus obliquus;

[0025] The volume ratio of the sterile inonotus birchii water extract, the northern schisandra chinensis extract and the tsaoko alcohol extract is (8-10): (3-5): (1-3).

[0026] Specific implementation method 2: The difference between this implementation method and specific implementation method 1 is that the volume ratio of the pretreated Inonotus obliquus to water in step S1 is 1: (8-12).

[0027] The other steps are the same as those in the first specific implementation.

[0028] Specific implementation method three: The difference between this implementation method and specific implementation method one or two is that the pretreated birch inonotus described in step S1 is processed according to the following steps: the fresh birch inonotus is dehydrated, and then the enzyme preparation is added, and then ultrasonic treatment is performed. After the ultrasonic treatment, it is dried and crushed, and then passed through an 80-mesh sieve to obtain the pretreated birch inonotus; the volume ratio of the dehydrated birch inonotus to the enzyme preparation is (50-80): 1, and the enzyme preparation is cellulase.

[0029] The other steps are the same as those in the first or second embodiment.

[0030] Specific embodiment 4: The difference between this embodiment and specific embodiments 1 to 3 is that the power of the microwave reactor in step S1 is 300-450W.

[0031] The other steps are the same as those in Specific Embodiments 1 to 3.

[0032] Specific implementation method five: The difference between this implementation method and specific implementation methods one to four is that the pore size of the microporous filtration membrane in step S1 and step S2 is 0.22 μm.

[0033] The other steps are the same as those in Specific Embodiments 1 to 4.

[0034] Specific embodiment 6: The difference between this embodiment and specific embodiments 1 to 5 is that the volume ratio of Schisandra chinensis chinensis to the polar solvent in step S2 is 1:(6-12).

[0035] The other steps are the same as those in Specific Embodiments 1 to 5.

[0036] Specific implementation method seven: The difference between this implementation method and specific implementation methods one to six is ​​that: the pressure in step S2 is 0.2-0.35Mpa; the ultrasonic power is 150-300W; the electric field strength of the high-voltage pulse electric field is 6-8kv / cm, and the number of pulses is 5-7.

[0037] The other steps are the same as those in Specific Embodiments 1 to 6.

[0038] Specific embodiment eight: This embodiment differs from specific embodiments one to seven in that: in step S2, purification is performed by using a macroporous adsorption resin or by alcohol precipitation after concentration.

[0039] The other steps are the same as those in Specific Embodiments 1 to 7.

[0040] Specific embodiment nine: The difference between this embodiment and specific embodiments one to eight is: the freeze-drying conditions in step S3: first adjust the temperature to -30℃~-50℃, pre-freeze for 1~2h; then adjust the temperature to -15℃~-25℃ under a vacuum degree of 40~60Pa, and freeze at a temperature of -15℃~-25℃ for 0.5~1.5h.

[0041] The other steps are the same as those in Specific Embodiments 1 to 8.

[0042] Specific embodiment ten: This embodiment provides an application of a composite freeze-dried powder based on Inonotus obliquus, and the application of the composite freeze-dried powder based on Inonotus obliquus in the preparation of anti-tumor, hypoglycemic, antihypertensive, antiviral and anti-aging drugs.

[0043] The following examples are used to verify the beneficial effects of the present invention:

[0044] Example 1: A method for preparing a composite freeze-dried powder of Inonotus obliquus, comprising the following steps:

[0045] Step S1: pretreatment of fresh Inonotus obliquus;

[0046] The fresh birch inonotus is dehydrated, and then an enzyme preparation is added, and then ultrasonic treatment is performed. After the ultrasonic treatment, the pretreated birch inonotus is dried and crushed, and then sieved through an 80-mesh sieve to obtain the pretreated birch inonotus; the volume ratio of the dehydrated birch inonotus to the enzyme preparation is 50:1, and the enzyme preparation is cellulase;

[0047] The fresh Inonotus birchii is purchased from common commercial products.

[0048] Step S2: Extraction of Inonotus obliquus;

[0049] 100 mL of the pretreated Inonotus birchii was mixed with 1 L of water, and then placed in a microwave reactor with a power of 400 W, heated to 100° C., and kept at 100° C. for 18 minutes to obtain an Inonotus birchii water extract; the Inonotus birchii water extract was sterilized and filtered using a 0.22 μm microporous filter membrane to obtain a sterile Inonotus birchii water extract;

[0050] Step S3: extracting Schisandra chinensis;

[0051] 100 mL of Schisandra chinensis was mixed with 1.2 L of polar solvent, heated to 50°C, extracted for 10 min under a pressure of 0.25 MPa, power of 250 W ultrasonic assistance, and a high-voltage pulse electric field with an electric field strength of 7 kv / cm and a pulse number of 6. After the extraction, it was purified using a macroporous adsorption resin, and then filtered using a 0.22 μm microporous filter membrane to obtain a Schisandra chinensis extract;

[0052] Step S4: preparation of composite freeze-dried powder;

[0053] The sterile inonotus birch water extract obtained in step S2, the Schisandra chinensis extract obtained in step S3 and the tsaoko alcohol extract are fully mixed and placed in a freeze dryer, first adjusted to -40°C and pre-frozen for 1.5 hours; then adjusted to -20°C under a vacuum degree of 50 Pa, and frozen at -20°C for 1.5 hours to obtain a composite freeze-dried powder based on inonotus birch; the volume ratio of the sterile inonotus birch water extract, the Schisandra chinensis extract and the tsaoko alcohol extract is 10:5:2.

[0054] Comparative Example 1: Step S3 is not performed, that is, the Schisandra chinensis extract is not added in Step S4; other experimental conditions are the same as those in Example 1.

[0055] Diabetic patients were selected to take the composite freeze-dried powder of Inonotus birchii in Example 1 as the experimental group; and the freeze-dried powder without adding Schisandra chinensis extract in Comparative Example 1 was taken as the control group. After 6 consecutive months of taking, the results showed that all the laboratory indicators of diabetes in the control group were significantly improved, and not only did the laboratory indicators of the experimental group return to normal, but also mild and severe patients did not need to take drugs to maintain a stable condition, indicating that the addition of Schisandra chinensis extract can effectively improve the patient's body immunity, and can play a significant effect of synergistic hypoglycemic and anti-sugar with Inonotus birchii.

[0056] Comparative Example 2: In step S4, no alcohol extract of Amomum villosum was added; other experimental conditions were the same as those in Example 1. The stability and activity of the effective components of the composite freeze-dried powder in Example 1 and Comparative Example 2 were investigated; the results showed that the stability of the composite freeze-dried powder of Inonotus obliquus in Example 1 was significantly higher than that in Comparative Example 2, and the biological activity of the composite freeze-dried powder in Example 1 was increased by 15-19% compared with that in Comparative Example 2, proving that the addition of alcohol extract of Amomum villosum significantly enhanced the antioxidant properties of the extract of Inonotus obliquus and the extract of Schisandra chinensis, and improved the biological activity of the composite freeze-dried powder.

[0057] Comparative Example 3: No enzyme preparation is added in step S1; other experimental conditions are the same as those in Example 1. The dissolution rates of the birch inonotus extracts of the two were calculated through experiments, and it was found that the dissolution rate of the birch inonotus extract in Example 1 was increased by about 5-8% compared with that in Comparative Example 3. Analysis showed that the enzyme preparation may have broken the pectin layer and cellulose layer on the surface of the birch inonotus, thereby increasing the dissolution amount of the birch inonotus extract.

[0058] Comparative Example 4: In step S2, a microwave reactor is not used, that is, normal water extraction is used; other experimental conditions are the same as those in Example 1. The experiment showed that the microwave reactor was used to assist extraction in Example 1, and the extraction time was shortened by about 30%, which greatly improved the extraction efficiency of Inonotus birchii.

[0059] Comparative Example 5: In step S3, a non-polar solvent is used for extraction; other experimental conditions are the same as those in Example 1. The solubility of schisandrin A of the two was measured by experiment; the measurement results showed that the solubility of schisandrin A in Example 1 was increased by about 30% compared with that in Comparative Example 5, which effectively verified the view that schisandrin A has a higher solubility in polar solvents and a lower solubility in non-polar solvents.

[0060] Comparative Example 6: In step S3, extraction was performed under normal pressure; other experimental conditions were the same as those in Example 1. The solubility of schisandrin A in the two samples was measured by experiments; the measurement results showed that the solubility of schisandrin A in Example 1 was increased by about 11% compared with that in Comparative Example 5, which fully proved that increasing the pressure would increase the contact between the solute and the solvent molecules, thereby increasing the solubility of the solute, i.e., schisandrin A.

[0061] Comparative Example 7: In step S3, ultrasound-assisted extraction is not used; other experimental conditions are the same as those in Example 1. The experiment shows that in Example 1, ultrasound-assisted extraction is used, and the average extraction rate of schisandrin A is 6.26 mg / g, which is much higher than 4.47 mg / g in Comparative Example 7.

[0062] Comparative Example 8: In step S3, the high-voltage pulse electric field method is not used for extraction; other experimental conditions are the same as those in Example 1. After measurement, the high-voltage pulse electric field method is used for extraction in Example 1, and the extraction rate of schisandrin A can be increased by about 19.2% compared with Comparative Example 8.

[0063] The relevant effective ingredients of the Inonotus obliquus composite freeze-dried powder prepared in Example 1 and similar Inonotus obliquus freeze-dried powder on the market were measured, and the specific data are shown in Table 1;

[0064] Table 1

[0065]

[0066]

[0067] As shown in Table 1, the composite freeze-dried powder of Inonotus birchii prepared by the process in Example 1 and compounding with Schisandra chinensis extract and Amomum villosum alcohol extract has a crude polysaccharide content increased by 27.67%, a Betulinum polysaccharide content increased by 23.2%, a β-glucan content increased by 14.28%, a triterpenoid content increased by 26.2%, and a Schisandrin A content increased by 40%, compared with similar freeze-dried powders of Inonotus birchii on the market. It has the potential to be used for preparing highly effective anti-tumor, hypoglycemic, hypotensive, antiviral and anti-aging drugs.

Claims

1. A method for preparing a composite freeze-dried powder of Inonotus obliquus, characterized in that The preparation method is carried out according to the following steps: Step S1: Extraction of Inonotus obliquus; The pretreated Inonotus birchii is mixed with water, and then placed in a microwave reactor, heated to 90-110° C., and kept warm at 90-110° C. for 15-20 minutes to obtain an Inonotus birchii water extract; the Inonotus birchii water extract is sterilized and filtered using a microporous filter membrane to obtain a sterile Inonotus birchii water extract; Step S2: extracting Schisandra chinensis; The schisandra chinensis is mixed with a polar solvent, heated to 45-55° C., and extracted for 5-10 minutes under pressure, ultrasound assistance and high-voltage pulse electric field conditions, purified after the extraction, and then filtered using a microporous filter membrane to obtain a schisandra chinensis extract; Step S3: preparation of composite freeze-dried powder; The sterile Inonotus obliquus water extract obtained in step S1, the Schisandra chinensis extract obtained in step S2 and the Amomum villosum alcohol extract are fully mixed, and then placed in a freeze dryer for freeze drying to obtain a composite freeze-dried powder based on Inonotus obliquus; The volume ratio of the sterile inonotus birchii water extract, the northern schisandra chinensis extract and the tsaoko alcohol extract is (8-10): (3-5): (1-3).

2. The method for preparing a composite freeze-dried powder of Inonotus obliquus according to claim 1, characterized in that The volume ratio of the pretreated Inonotus obliquus to water in step S1 is 1:(8-12).

3. A method for preparing a composite freeze-dried powder based on Inonotus obliquus according to claim 1 or 2, characterized in that The pretreated Inonotus birchii described in step S1 is treated according to the following steps: the fresh Inonotus birchii is dehydrated, then an enzyme preparation is added, and then ultrasonic treatment is performed, and after the ultrasonic treatment, it is dried and crushed, and then passed through an 80-mesh sieve to obtain the pretreated Inonotus birchii; the volume ratio of the dehydrated Inonotus birchii to the enzyme preparation is (50-80):1, and the enzyme preparation is cellulase.

4. The method for preparing a composite freeze-dried powder of Inonotus obliquus according to claim 1, characterized in that The power of the microwave reactor in step S1 is 300-450W.

5. The method for preparing a composite freeze-dried powder of Inonotus obliquus according to claim 1, characterized in that The pore size of the microporous filtration membrane in step S1 and step S2 is 0.22 μm.

6. The method for preparing a composite freeze-dried powder of Inonotus obliquus according to claim 1, characterized in that The volume ratio of Schisandra chinensis chinensis to the polar solvent in step S2 is 1:(6-12).

7. The method for preparing a composite freeze-dried powder based on Inonotus obliquus according to claim 1, characterized in that In step S2, the pressure is 0.2-0.35 MPa; the ultrasonic power is 150-300 W; the electric field strength of the high-voltage pulse electric field is 6-8 kV / cm, and the number of pulses is 5-7.

8. The method for preparing a composite freeze-dried powder of Inonotus obliquus according to claim 1, characterized in that In step S2, the purification is performed by using a macroporous adsorption resin or by concentration followed by alcohol precipitation.

9. The method for preparing a composite freeze-dried powder of Inonotus obliquus according to claim 1, characterized in that The freeze-drying conditions in step S3 are as follows: first adjust the temperature to -30°C to -50°C and pre-freeze for 1 to 2 hours; then adjust the temperature to -15°C to -25°C under a vacuum degree of 40 to 60 Pa, and freeze at a temperature of -15°C to -25°C for 0.5 to 1.5 hours.

10. An application of a composite freeze-dried powder based on Inonotus obliquus prepared by the method according to any one of claims 1 to 9, characterized in that The application of the composite freeze-dried powder based on Inonotus obliquus in the preparation of anti-tumor, hypoglycemic, hypotensive, antiviral and anti-aging drugs.

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