CN112126562A - Gorgon fruit extract-based health care wine for reducing blood sugar and inhibiting cancer cell proliferation - Google Patents
Gorgon fruit extract-based health care wine for reducing blood sugar and inhibiting cancer cell proliferation Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
- C12G3/024—Preparation of other alcoholic beverages by fermentation of fruits other than botanical genus Vitis
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/06—Fungi, e.g. yeasts
- A61K36/07—Basidiomycota, e.g. Cryptococcus
- A61K36/074—Ganoderma
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/62—Nymphaeaceae (Water-lily family)
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/73—Rosaceae (Rose family), e.g. strawberry, chokeberry, blackberry, pear or firethorn
- A61K36/736—Prunus, e.g. plum, cherry, peach, apricot or almond
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/88—Liliopsida (monocotyledons)
- A61K36/896—Liliaceae (Lily family), e.g. daylily, plantain lily, Hyacinth or narcissus
- A61K36/8967—Lilium, e.g. tiger lily or Easter lily
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
- C12G3/026—Preparation of other alcoholic beverages by fermentation with health-improving ingredients, e.g. flavonoids, flavones, polyphenols or polysaccharides, added before or during the fermentation stage; with flavouring ingredients added before or during the fermentation stage
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention provides a gordon euryale seed extract-based health care wine for reducing blood sugar and inhibiting cancer cell proliferation, which is prepared by crushing gordon euryale seed into gordon euryale seed powder, sequentially extracting the gordon euryale seed powder with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, and concentrating and drying the extracting solution to obtain a gordon euryale seed extract; then crushing the lily slices into lily powder, removing cores of fresh green plums and crushing into green plum pulp, then adding the lily powder and residues into the green plum pulp, homogenizing, inoculating ganoderma lucidum strains, culturing and crushing to obtain a culture; dispersing the culture in deionized water by ultrasonic wave, adding 1, 3-propylene glycol, and processing to obtain extract; and finally, adding the gordon euryale seed extract and the second extract into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting and performing post-treatment to obtain the health-care wine which has the double effects of reducing blood sugar and inhibiting cancer cell proliferation.
Description
Technical Field
The invention relates to the technical field, in particular to gorgon euryale seed extract-based health care wine for reducing blood sugar and inhibiting cancer cell proliferation.
Background
In recent years, the continuous and rapid increase of patients with hyperglycemia or diabetes has become one of the main health problems of human beings. Studies have shown that the main characteristic of such patients is excessive blood glucose concentration, which is a significant cause of diabetic complications, and thus, control of blood glucose concentration is very important for prevention and treatment of such diseases. Carbohydrates such as starch are one of the main components in human diet and also important sources of human energy. After digestion and absorption, starch enters the blood in the form of glucose, and as a result, the blood sugar is increased. The digestion of starch needs to be catalyzed by enzymes such as alpha-amylase and alpha-glucosidase, so that the activity of the enzymes related to the starch digestion is inhibited, the digestion and absorption of the starch can be delayed, and the postprandial hyperglycemia can be inhibited.
Cancer is one of the leading causes of death. In body tissues, cell differentiation is a physiological process, and under normal physiological conditions, cell proliferation and cell death are in a certain balance. The external environment such as ray, air pollution and the like can cause the mutation of human DNA, thereby breaking the normal regulation and control process of the human body to cause the generation of cancer. At both the cellular and genetic level, cells undergo uncontrolled differentiation during the development of cancer, forming malignant tumors and spreading to other parts of the body. Therefore, the inhibition of cancer cell proliferation has excellent health care significance.
Patent CN106244371B discloses a polygonatum sibiricum grape health wine for reducing blood sugar, which is prepared by fermenting grapes serving as raw materials with yeast, wherein traditional Chinese medicine components such as rhizoma polygonati powder, radix puerariae powder and dark plum are added in the fermentation process, so that the polygonatum sibiricum grape health wine has a certain effect of reducing blood sugar. However, the health wine has limited hypoglycemic effect and no effect of inhibiting cancer cell proliferation.
Disclosure of Invention
The invention aims to provide a gorgon euryale seed extract-based health care wine for reducing blood sugar and inhibiting cancer cell proliferation, which has the dual effects of reducing blood sugar and inhibiting cancer cell proliferation.
In order to achieve the purpose, the invention is realized by the following scheme:
a preparation method of health wine based on semen euryales extract for reducing blood sugar and inhibiting cancer cell proliferation comprises the following specific steps:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating and drying the extracting solution to obtain a gordon euryale seed extract, and allowing the residues to enter the step (2);
(2) then crushing the lily slices into lily powder, removing kernels of the fresh green plums and crushing the fresh green plums into green plum pulp, then adding the lily powder and the residues obtained in the step (1) into the green plum pulp, homogenizing, inoculating ganoderma lucidum strains, culturing and crushing to obtain a culture;
(3) dispersing the culture in deionized water by ultrasonic waves, adding 1, 3-propylene glycol, uniformly stirring, standing for the first time, concentrating under reduced pressure, adding absolute ethyl alcohol until the volume content of the ethyl alcohol in the system is 60-70%, uniformly stirring, standing for the second time, centrifuging, taking supernatant, concentrating and drying to obtain a second extract;
(4) and (3) finally, adding the gordon euryale seed extract obtained in the step (1) and the secondary extract obtained in the step (3) into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting, and performing post-treatment to obtain the health-care wine.
Preferably, in the step (1), the particle size of the gordon euryale seed powder is 100-200 meshes.
Preferably, in the step (1), the specific extraction method comprises the following steps in parts by weight: firstly, adding 1 part of gordon euryale seed powder into 5-8 parts of deionized water, and carrying out ultrasonic oscillation extraction at 50-60 ℃ and 300-500W for 50-80 minutes to obtain an extracting solution I and a residue I; and then adding the residue I into 4-6 parts of absolute ethyl alcohol, carrying out ultrasonic oscillation extraction at 50-60 ℃ and 300-500W for 70-90 minutes to obtain an extracting solution II and a residue II, combining the extracting solution I and the extracting solution II to obtain an extracting solution, and combining the residue I and the residue II to obtain a residue.
Preferably, in the step (2), the particle size of the lily powder is 100-200 meshes.
Preferably, in the step (2), the mass ratio of the medicinal lily powder, the residue, the green plum pulp and the ganoderma lucidum strain is 1: 5-7: 8-10: 0.8 to 1.
Preferably, in the step (2), the homogenization treatment is realized by adopting ultrahigh pressure microjet, and the specific process conditions are as follows: treating for 2-3 times under 200-250 MPa.
Preferably, in the step (2), the culture conditions are as follows: culturing for 45-60 days at 25-30 ℃ and 90-95% humidity; after the culture, Ganoderma mycelium and Ganoderma fruiting body are obtained, and are pulverized to particle size of below 10 μm to obtain culture.
Preferably, in the step (3), the mass ratio of the culture, the deionized water and the 1, 3-propanediol is 1: 5-7: 0.06-0.08.
Preferably, in the step (3), the first standing time is 2-4 hours, and the second standing time is 5-8 hours.
Preferably, in the step (4), the mass ratio of the gordon euryale seed extract to the deionized water to the high-activity yeast liquid is 1: 8-10: 0.05 to 0.08.
Preferably, in the step (4), the high-activity yeast solution is obtained by adding high-activity dry yeast into 35 ℃ water with the weight 10-15 times that of the high-activity dry yeast solution, and dissolving and activating the high-activity dry yeast solution for 20-30 minutes.
Preferably, in the step (4), the fermentation process conditions are as follows: fermenting for 3-4 days at 18-25 ℃.
Preferably, in the step (4), the post-treatment comprises: standing for clarification, filtering, transferring the obtained filtrate into an aging tank, aging for 100-120 days, filtering, and pasteurizing.
The gordon euryale seed extract-based health care wine for reducing blood sugar and inhibiting cancer cell proliferation is obtained by the preparation method.
Compared with the prior art, the invention has the beneficial effects that:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating the extracting solution, and drying to obtain a gordon euryale seed extract; then crushing the lily slices into lily powder, removing cores of fresh green plums and crushing into green plum pulp, then adding the lily powder and residues into the green plum pulp, homogenizing, inoculating ganoderma lucidum strains, culturing and crushing to obtain a culture; dispersing the culture in deionized water by ultrasonic waves, adding 1, 3-propylene glycol, uniformly stirring, standing for the first time, concentrating under reduced pressure, adding absolute ethyl alcohol until the volume content of the ethyl alcohol in the system is 60-70%, uniformly stirring, standing for the second time, centrifuging, taking supernatant, concentrating and drying to obtain a second extract; and finally, adding the gordon euryale seed extract and the second extract into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting and performing post-treatment to obtain the health-care wine which has the double effects of reducing blood sugar and inhibiting cancer cell proliferation.
(2) The gorgon fruit contains components such as polyphenol, polysaccharide, thiamine and the like, which are beneficial to reducing the blood sugar level, wherein the polyphenol can play a role in controlling the release of blood sugar; the polysaccharide promotes insulin secretion, regulates key enzyme activity and increases insulin sensitivity; thiamine can maintain normal carbohydrate metabolism and control blood glucose levels. When the gordon euryale seed is extracted, deionized water and absolute ethyl alcohol are sequentially used for extraction, so that water-soluble polysaccharide, thiamine, alcohol-soluble polyphenol and amylase inhibitory components can be extracted, and the blood sugar reducing effect is improved. In addition, polyphenol can inhibit cancer cell proliferation, and the gorgon euryale seed extract obtained by the extraction of the invention contains more glycopeptides and can inhibit cancer cell proliferation together with polyphenol.
(3) The residue of the invention is also mixed with the medicinal lily, green plum and the like, and then inoculated with the ganoderma lucidum strain to realize re-extraction, the medicinal lily contains cells for repairing pancreatic islets, the insulin secretion is increased, the blood sugar reducing effect is further improved, and the green plum, the ganoderma lucidum and the like reduce hepatic glycogen to play a role in reducing blood sugar. The polyphenol contained in Bulbus Lilii, mume fructus, and Ganoderma can further inhibit cancer cell proliferation. In addition, the invention inoculates the ganoderma lucidum strain in the mixture prepared by the medicinal lily powder, green plum pulp and residues, and generates more beneficial substances such as glycopeptide and the like along with the growth of ganoderma lucidum in the culture process, thereby effectively improving the effects of reducing blood sugar and inhibiting cancer cell proliferation of the product. The addition of 1, 3-propanediol helps to enrich these beneficial components, further improving the health care effect.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The ganoderma lucidum strain is purchased from Beijing Beinanna Chuanglian union Biotechnology research institute.
Example 1
A preparation method of health wine based on semen euryales extract for reducing blood sugar and inhibiting cancer cell proliferation comprises the following specific steps:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating and drying the extracting solution to obtain a gordon euryale seed extract, and allowing the residues to enter the step (2);
(2) then crushing the lily slices into lily powder, removing kernels of the fresh green plums and crushing the fresh green plums into green plum pulp, then adding the lily powder and the residues obtained in the step (1) into the green plum pulp, homogenizing, inoculating ganoderma lucidum strains, culturing and crushing to obtain a culture;
(3) dispersing the culture in deionized water by ultrasonic wave, adding 1, 3-propylene glycol, stirring, standing for the first time, concentrating under reduced pressure, adding anhydrous ethanol until the volume content of ethanol in the system is 60%, stirring, standing for the second time, centrifuging, collecting supernatant, concentrating, and drying to obtain extract again;
(4) and (3) finally, adding the gordon euryale seed extract obtained in the step (1) and the secondary extract obtained in the step (3) into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting, and performing post-treatment to obtain the health-care wine.
In the step (1), the particle size of the gordon euryale seed powder is 100 meshes.
In the step (1), the specific extraction method is as follows: firstly, adding 1kg of gordon euryale seed powder into 8kg of deionized water, and carrying out ultrasonic oscillation extraction at 50 ℃ and 500W for 50 minutes to obtain an extracting solution I and a residue I; and then adding the residue I into 6kg of absolute ethyl alcohol, carrying out ultrasonic oscillation extraction at 50 ℃ and 500W for 70 minutes to obtain an extracting solution II and a residue II, combining the extracting solution I and the extracting solution II to obtain an extracting solution, and combining the residue I and the residue II to obtain a residue.
In the step (2), the particle size of the lily powder is 200 meshes.
In the step (2), the mass ratio of the medicinal lily powder, the residue, the green plum pulp and the ganoderma lucidum strain is 1: 5: 10: 0.8.
in the step (2), homogenization treatment is realized by adopting ultrahigh-pressure microjet, and the specific process conditions are as follows: treating under 250MPa for 2 times.
In the step (2), the culture conditions are as follows: culturing at 30 deg.C and 90% humidity for 60 days; after the culture, Ganoderma mycelium and Ganoderma fruiting body are obtained, and are pulverized to particle size of below 10 μm to obtain culture.
In the step (3), the mass ratio of the culture, the deionized water and the 1, 3-propylene glycol is 1: 5: 0.08.
in the step (3), the first standing time is 2 hours, and the second standing time is 8 hours.
In the step (4), the mass ratio of the gordon euryale seed extract, the deionized water and the high-activity yeast liquid is 1: 8: 0.08.
in the step (4), the high-activity yeast solution is obtained by adding high-activity dry yeast into 10 times of 35 ℃ water by weight, and dissolving and activating for 30 minutes.
In the step (4), the fermentation process conditions are as follows: fermenting at 18 deg.C for 4 days.
In the step (4), the post-processing comprises: standing for clarifying, filtering, aging in aging tank for 100 days, filtering, and pasteurizing.
Example 2
A preparation method of health wine based on semen euryales extract for reducing blood sugar and inhibiting cancer cell proliferation comprises the following specific steps:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating and drying the extracting solution to obtain a gordon euryale seed extract, and allowing the residues to enter the step (2);
(2) then crushing the lily slices into lily powder, removing kernels of the fresh green plums and crushing the fresh green plums into green plum pulp, then adding the lily powder and the residues obtained in the step (1) into the green plum pulp, homogenizing, inoculating ganoderma lucidum strains, culturing and crushing to obtain a culture;
(3) dispersing the culture in deionized water by ultrasonic wave, adding 1, 3-propylene glycol, stirring, standing for the first time, concentrating under reduced pressure, adding anhydrous ethanol until the volume content of ethanol in the system is 70%, stirring, standing for the second time, centrifuging, collecting supernatant, concentrating, and drying to obtain extract again;
(4) and (3) finally, adding the gordon euryale seed extract obtained in the step (1) and the secondary extract obtained in the step (3) into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting, and performing post-treatment to obtain the health-care wine.
In the step (1), the particle size of the gordon euryale seed powder is 100 meshes.
In the step (1), the specific extraction method is as follows: firstly, adding 1kg of gordon euryale seed powder into 5kg of deionized water, and carrying out ultrasonic oscillation extraction at 60 ℃ and 300W for 80 minutes to obtain an extracting solution I and a residue I; and then adding the residue I into 4kg of absolute ethyl alcohol, carrying out ultrasonic oscillation extraction at 60 ℃ and 300W for 90 minutes to obtain an extracting solution II and a residue II, combining the extracting solution I and the extracting solution II to obtain an extracting solution, and combining the residue I and the residue II to obtain a residue.
In the step (2), the particle size of the lily powder is 100 meshes.
In the step (2), the mass ratio of the medicinal lily powder, the residue, the green plum pulp and the ganoderma lucidum strain is 1: 7: 8: 1.
in the step (2), homogenization treatment is realized by adopting ultrahigh-pressure microjet, and the specific process conditions are as follows: treating under 200MPa for 3 times.
In the step (2), the culture conditions are as follows: culturing at 25 deg.C and 95% humidity for 45 days; after the culture, Ganoderma mycelium and Ganoderma fruiting body are obtained, and are pulverized to particle size of below 10 μm to obtain culture.
In the step (3), the mass ratio of the culture, the deionized water and the 1, 3-propylene glycol is 1: 7: 0.06.
in the step (3), the first standing time is 4 hours, and the second standing time is 5 hours.
In the step (4), the mass ratio of the gordon euryale seed extract, the deionized water and the high-activity yeast liquid is 1: 10: 0.05.
in the step (4), the high-activity yeast solution is obtained by adding high-activity dry yeast into 15 times of 35 ℃ water by weight, and dissolving and activating for 20 minutes.
In the step (4), the fermentation process conditions are as follows: fermenting at 25 deg.C for 3 days.
In the step (4), the post-processing comprises: standing for clarifying, filtering, aging for 120 days, filtering, and pasteurizing.
Example 3
A preparation method of health wine based on semen euryales extract for reducing blood sugar and inhibiting cancer cell proliferation comprises the following specific steps:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating and drying the extracting solution to obtain a gordon euryale seed extract, and allowing the residues to enter the step (2);
(2) then crushing the lily slices into lily powder, removing kernels of the fresh green plums and crushing the fresh green plums into green plum pulp, then adding the lily powder and the residues obtained in the step (1) into the green plum pulp, homogenizing, inoculating ganoderma lucidum strains, culturing and crushing to obtain a culture;
(3) dispersing the culture in deionized water by ultrasonic wave, adding 1, 3-propylene glycol, stirring, standing for the first time, concentrating under reduced pressure, adding anhydrous ethanol until the volume content of ethanol in the system is 65%, stirring, standing for the second time, centrifuging, collecting supernatant, concentrating, and drying to obtain extract again;
(4) and (3) finally, adding the gordon euryale seed extract obtained in the step (1) and the secondary extract obtained in the step (3) into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting, and performing post-treatment to obtain the health-care wine.
In the step (1), the particle size of the gordon euryale seed powder is 200 meshes.
In the step (1), the specific extraction method is as follows: firstly, adding 1kg of gordon euryale seed powder into 7kg of deionized water, and carrying out ultrasonic oscillation extraction at 55 ℃ and 400W for 70 minutes to obtain an extracting solution I and a residue I; and then adding the residue I into 5kg of absolute ethyl alcohol, carrying out ultrasonic oscillation extraction at 55 ℃ and 400W for 80 minutes to obtain an extracting solution II and a residue II, combining the extracting solution I and the extracting solution II to obtain an extracting solution, and combining the residue I and the residue II to obtain a residue.
In the step (2), the particle size of the lily powder is 200 meshes.
In the step (2), the mass ratio of the medicinal lily powder, the residue, the green plum pulp and the ganoderma lucidum strain is 1: 6: 9: 0.9.
in the step (2), homogenization treatment is realized by adopting ultrahigh-pressure microjet, and the specific process conditions are as follows: treating for 2 times under the condition of 220 MPa.
In the step (2), the culture conditions are as follows: culturing at 28 deg.C and 92% humidity for 55 days; after the culture, Ganoderma mycelium and Ganoderma fruiting body are obtained, and are pulverized to particle size of below 10 μm to obtain culture.
In the step (3), the mass ratio of the culture, the deionized water and the 1, 3-propylene glycol is 1: 6: 0.07.
in the step (3), the first standing time is 3 hours, and the second standing time is 6 hours.
In the step (4), the mass ratio of the gordon euryale seed extract, the deionized water and the high-activity yeast liquid is 1: 9: 0.07.
in the step (4), the high-activity yeast solution is obtained by adding high-activity dry yeast into 35 ℃ water with the weight being 12 times of that of the high-activity dry yeast, and dissolving and activating the high-activity dry yeast for 25 minutes.
In the step (4), the fermentation process conditions are as follows: fermenting at 20 deg.C for 4 days.
In the step (4), the post-processing comprises: standing for clarifying, filtering, aging for 115 days in aging tank, filtering, and pasteurizing.
Comparative example 1
A preparation method of health wine based on semen euryales extract for reducing blood sugar and inhibiting cancer cell proliferation comprises the following specific steps:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating and drying the extracting solution to obtain a gordon euryale seed extract, and allowing the residues to enter the step (2);
(2) and (2) adding the gordon euryale seed extract obtained in the step (1) into deionized water, uniformly oscillating by ultrasonic waves, adding a high-activity yeast solution, fermenting, and performing post-treatment to obtain the health wine.
In the step (1), the particle size of the gordon euryale seed powder is 100 meshes.
In the step (1), the specific extraction method is as follows: firstly, adding 1kg of gordon euryale seed powder into 8kg of deionized water, and carrying out ultrasonic oscillation extraction at 50 ℃ and 500W for 50 minutes to obtain an extracting solution I and a residue I; and then adding the residue I into 6kg of absolute ethyl alcohol, carrying out ultrasonic oscillation extraction at 50 ℃ and 500W for 70 minutes to obtain an extracting solution II and a residue II, combining the extracting solution I and the extracting solution II to obtain an extracting solution, and combining the residue I and the residue II to obtain a residue.
In the step (2), the mass ratio of the gordon euryale seed extract, the deionized water and the high-activity yeast liquid is 1: 8: 0.08.
in the step (2), the high-activity yeast solution is obtained by adding high-activity dry yeast into 10 times of 35 ℃ water by weight, and dissolving and activating for 30 minutes.
In the step (2), the fermentation process conditions are as follows: fermenting at 18 deg.C for 4 days.
In the step (2), the post-treatment comprises: standing for clarifying, filtering, aging in aging tank for 100 days, filtering, and pasteurizing.
Comparative example 2
A preparation method of health wine based on semen euryales extract for reducing blood sugar and inhibiting cancer cell proliferation comprises the following specific steps:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating and drying the extracting solution to obtain a gordon euryale seed extract, and allowing the residues to enter the step (2);
(2) then crushing the lily tablets into lily powder, removing kernels of the fresh green plums and crushing the fresh green plums into green plum pulp, adding the lily powder, the residue obtained in the step (1) and ganoderma lucidum powder (purchased from Guanyu agriculture Co., Ltd. in Guangxi county) into the green plum pulp, and homogenizing to obtain a mixture;
(3) dispersing the mixture in deionized water by ultrasonic wave, adding 1, 3-propylene glycol, stirring, standing for the first time, concentrating under reduced pressure, adding anhydrous ethanol until the volume content of ethanol in the system is 60%, stirring, standing for the second time, centrifuging, collecting supernatant, concentrating, and drying to obtain extract again;
(4) and (3) finally, adding the gordon euryale seed extract obtained in the step (1) and the secondary extract obtained in the step (3) into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting, and performing post-treatment to obtain the health-care wine.
In the step (1), the particle size of the gordon euryale seed powder is 100 meshes.
In the step (1), the specific extraction method is as follows: firstly, adding 1kg of gordon euryale seed powder into 8kg of deionized water, and carrying out ultrasonic oscillation extraction at 50 ℃ and 500W for 50 minutes to obtain an extracting solution I and a residue I; and then adding the residue I into 6kg of absolute ethyl alcohol, carrying out ultrasonic oscillation extraction at 50 ℃ and 500W for 70 minutes to obtain an extracting solution II and a residue II, combining the extracting solution I and the extracting solution II to obtain an extracting solution, and combining the residue I and the residue II to obtain a residue.
In the step (2), the particle size of the lily powder is 200 meshes.
In the step (2), the mass ratio of the medicinal lily powder, the residue, the green plum pulp and the lucid ganoderma powder is 1: 5: 10: 0.8.
in the step (2), homogenization treatment is realized by adopting ultrahigh-pressure microjet, and the specific process conditions are as follows: treating under 250MPa for 2 times.
In the step (3), the mass ratio of the mixture, the deionized water and the 1, 3-propylene glycol is 1: 5: 0.08.
in the step (3), the first standing time is 2 hours, and the second standing time is 8 hours.
In the step (4), the mass ratio of the gordon euryale seed extract, the deionized water and the high-activity yeast liquid is 1: 8: 0.08.
in the step (4), the high-activity yeast solution is obtained by adding high-activity dry yeast into 10 times of 35 ℃ water by weight, and dissolving and activating for 30 minutes.
In the step (4), the fermentation process conditions are as follows: fermenting at 18 deg.C for 4 days.
In the step (4), the post-processing comprises: standing for clarifying, filtering, aging in aging tank for 100 days, filtering, and pasteurizing.
Comparative example 3
A preparation method of health wine based on semen euryales extract for reducing blood sugar and inhibiting cancer cell proliferation comprises the following specific steps:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating and drying the extracting solution to obtain a gordon euryale seed extract, and allowing the residues to enter the step (2);
(2) then crushing the lily slices into lily powder, removing kernels of the fresh green plums and crushing the fresh green plums into green plum pulp, then adding the lily powder and the residues obtained in the step (1) into the green plum pulp, homogenizing, inoculating ganoderma lucidum strains, culturing and crushing to obtain a culture;
(3) and (3) finally, adding the gordon euryale seed extract obtained in the step (1) and the culture obtained in the step (2) into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting, and performing post-treatment to obtain the health wine.
In the step (1), the particle size of the gordon euryale seed powder is 100 meshes.
In the step (1), the specific extraction method is as follows: firstly, adding 1kg of gordon euryale seed powder into 8kg of deionized water, and carrying out ultrasonic oscillation extraction at 50 ℃ and 500W for 50 minutes to obtain an extracting solution I and a residue I; and then adding the residue I into 6kg of absolute ethyl alcohol, carrying out ultrasonic oscillation extraction at 50 ℃ and 500W for 70 minutes to obtain an extracting solution II and a residue II, combining the extracting solution I and the extracting solution II to obtain an extracting solution, and combining the residue I and the residue II to obtain a residue.
In the step (2), the particle size of the lily powder is 200 meshes.
In the step (2), the mass ratio of the medicinal lily powder, the residue, the green plum pulp and the ganoderma lucidum strain is 1: 5: 10: 0.8.
in the step (2), homogenization treatment is realized by adopting ultrahigh-pressure microjet, and the specific process conditions are as follows: treating under 250MPa for 2 times.
In the step (2), the culture conditions are as follows: culturing at 30 deg.C and 90% humidity for 60 days; after the culture, Ganoderma mycelium and Ganoderma fruiting body are obtained, and are pulverized to particle size of below 10 μm to obtain culture.
In the step (3), the mass ratio of the gordon euryale seed extract to the deionized water to the high-activity yeast liquid is 1: 8: 0.08.
in the step (3), the high-activity yeast solution is obtained by adding high-activity dry yeast into 10 times of 35 ℃ water by weight, and dissolving and activating for 30 minutes.
In the step (3), the fermentation process conditions are as follows: fermenting at 18 deg.C for 4 days.
In the step (3), the post-processing comprises: standing for clarifying, filtering, aging in aging tank for 100 days, filtering, and pasteurizing.
Test examples
1. Investigation of hypoglycemic Effect
Healthy adult mice are selected, after 1 day of adaptation, 100 animals are randomly selected to be fasted for 4 hours, and fasting blood glucose is measured to serve as the basic blood glucose value of the batch of animals. The animals were then fasted for 24 hours (free drinking), molded with alloxan (freshly prepared before use) and injected at a dose of 48mg/kg body weight. After 5 days, the animals are fasted for 4 hours, the blood sugar is measured, and the blood sugar value is 10-25 mmol/L, which is the animal success in the hyperglycemia model.
70 hyperglycemic model animals are selected, and are randomly and averagely divided into 7 groups, and 1 model control group and 6 test groups are randomly selected (the difference between the groups is not more than 1.1 mmol/L). The test groups are respectively given the health-care wine of examples 1-3 and comparative examples 1-3, the administration mode of the health-care wine is oral administration, the volume of the administered health-care wine is 0.4mL/20g of body weight, the model control group is given deionized water with the same volume for 20 days continuously, blood is taken from the orbit to measure the blood sugar, and the blood sugar values (average) of animals in each group before and after administration are compared. The results are shown in Table 1.
TABLE 1 examination of hypoglycemic Effect
| Before administration (blood sugar level, mmol/L) | After administration (blood sugar level, mmol/L) | |
| Example 1 | 17.42 | 6.23 |
| Example 2 | 17.48 | 6.29 |
| Example 3 | 17.41 | 6.08 |
| Comparative example 1 | 17.38 | 11.89 |
| Comparative example 2 | 17.45 | 10.03 |
| Comparative example 3 | 17.46 | 8.72 |
| Model control group | 17.39 | 15.11 |
As can be seen from Table 1, the health-care wines of examples 1-3 have a good blood sugar-reducing effect compared with the model control group.
Comparative example 1 the gordon euryale seed extract is obtained after extraction with deionized water and absolute ethyl alcohol in the step (1), and the residue is discarded, and the steps (2) and (3) are omitted; comparative example 2 traditional Chinese medicine lily powder and green plum syrup were directly mixed with ganoderma lucidum powder without culturing; in a comparative example 3, the step (3) is omitted, the blood sugar reducing effect of the obtained health care wine is obviously poor, the residue after the primary extraction of the gordon euryale seed still contains a large amount of beneficial components, the production of the beneficial components is more facilitated by inoculating the ganoderma lucidum strain into the mixture of the medicinal lily powder and the green plum pulp for culture, and the further enrichment of the beneficial components is facilitated by the 1, 3-propylene glycol.
2. Investigation of cancer cell inhibition
Human ovarian carcinoma cells OVCAR-3 (purchased from Shanghai Haishen industries, Ltd.) were cultured in RPMI 1640 medium containing 10% calf serum and 1% double antibody (100U/mL penicillin and 100U/mL streptomycin) at 37 ℃ in 5% CO2Saturated humidity CO2Culturing in an incubator, and taking cells in logarithmic growth phase for experiment.
The influence of the health care wine obtained in the examples 1-3 or the comparative examples 1-3 on the cell proliferation of the human ovarian cancer cell OVCAR-3 is determined by an MTT colorimetric method, and the specific method is as follows:
the health wine is dissolved in RPMI 1640 culture medium containing 10% calf serum and 1% double antibody (100U/mL penicillin and 100U/mL streptomycin) to obtain solutions with health wine concentration of 0.8, 1.6, 3.2, 6.25, 12.5 and 25mg/mL, which are used for MTT dosing experiment.
Taking cells in logarithmic growth phase at 2X 10 per well4The cells were seeded in a 96-well culture plate, 100. mu.L of cell suspension was added to each well, cultured in a cell incubator for 24 hours, and the cell adhesion state was examined. Well-adherent 96-well plates of cells can be subjected to the next dosing experiment. The medium was aspirated from all wells, the solution was added to each well, and the control group was added with no health wine, but only R containing 10% calf serum, 1% diabody (100U/mL penicillin and 100U/mL streptomycin)PMI 1640 medium. Each group was treated for 120 hours with 4 identical cell wells per concentration. After the completion of the incubation, 25. mu.L of MTT solution at a concentration of 5mg/mL was added to each well, and the mixture was placed in an incubator for further incubation for 4 hours, then 100. mu.L of 10% SDS-0.1MHCl solution was added to each well, and the mixture was placed in the incubator overnight to dissolve the bluish purple precipitate sufficiently, and the absorbance at 570nm was measured with a microplate reader.
The result of the health care wine on the proliferation inhibition effect of human ovarian cancer cell OVCAR-3 is shown in Table 2.
TABLE 2 examination of the Effect of inhibiting cancer cells
| IC50(mg/mL) | |
| Example 1 | 3.1 |
| Example 2 | 3.0 |
| Example 3 | 2.8 |
| Comparative example 1 | 8.8 |
| Comparative example 2 | 6.5 |
| Comparative example 3 | 4.6 |
As can be seen from Table 2, the health liquors of examples 1 to 3 had IC' s50Low content of cancer cells, and good effect of inhibiting cancer cells.
Comparative example 1 the gordon euryale seed extract is obtained after extraction with deionized water and absolute ethyl alcohol in the step (1), and the residue is discarded, and the steps (2) and (3) are omitted; comparative example 2 traditional Chinese medicine lily powder and green plum syrup were directly mixed with ganoderma lucidum powder without culturing; in a comparative example 3, the step (3) is omitted, the cancer cell proliferation inhibition effect of the obtained health care wine is obviously poor, the residue after the primary extraction of the gordon euryale seed still contains a large amount of beneficial components, the production of the beneficial components is more facilitated by inoculating the ganoderma lucidum strain into the mixture of the medicinal lily powder and the green plum pulp for culture, and the further enrichment of the beneficial components is facilitated by the 1, 3-propylene glycol.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (10)
1. A preparation method of health care wine for reducing blood sugar and inhibiting cancer cell proliferation based on a gordon euryale seed extract is characterized by comprising the following specific steps:
(1) crushing gordon euryale seeds into gordon euryale seed powder, sequentially extracting with deionized water and absolute ethyl alcohol to obtain an extracting solution and residues, concentrating and drying the extracting solution to obtain a gordon euryale seed extract, and allowing the residues to enter the step (2);
(2) then crushing the lily slices into lily powder, removing kernels of the fresh green plums and crushing the fresh green plums into green plum pulp, then adding the lily powder and the residues obtained in the step (1) into the green plum pulp, homogenizing, inoculating ganoderma lucidum strains, culturing and crushing to obtain a culture;
(3) dispersing the culture in deionized water by ultrasonic waves, adding 1, 3-propylene glycol, uniformly stirring, standing for the first time, concentrating under reduced pressure, adding absolute ethyl alcohol until the volume content of the ethyl alcohol in the system is 60-70%, uniformly stirring, standing for the second time, centrifuging, taking supernatant, concentrating and drying to obtain a second extract;
(4) and (3) finally, adding the gordon euryale seed extract obtained in the step (1) and the secondary extract obtained in the step (3) into deionized water, uniformly oscillating by ultrasonic waves, adding high-activity yeast liquid, fermenting, and performing post-treatment to obtain the health-care wine.
2. The preparation method according to claim 1, wherein in the step (1), the specific method of extraction is as follows in parts by weight: firstly, adding 1 part of gordon euryale seed powder into 5-8 parts of deionized water, and carrying out ultrasonic oscillation extraction at 50-60 ℃ and 300-500W for 50-80 minutes to obtain an extracting solution I and a residue I; and then adding the residue I into 4-6 parts of absolute ethyl alcohol, carrying out ultrasonic oscillation extraction at 50-60 ℃ and 300-500W for 70-90 minutes to obtain an extracting solution II and a residue II, combining the extracting solution I and the extracting solution II to obtain an extracting solution, and combining the residue I and the residue II to obtain a residue.
3. The preparation method according to claim 1, wherein in the step (2), the mass ratio of the medicinal lily powder, the residue, the green plum pulp and the ganoderma lucidum strain is 1: 5-7: 8-10: 0.8 to 1.
4. The preparation method according to claim 1, wherein in the step (2), the homogenization treatment is realized by adopting ultrahigh pressure micro-jet, and the specific process conditions are as follows: treating for 2-3 times under 200-250 MPa.
5. The method according to claim 1, wherein in the step (2), the culture conditions are: culturing for 45-60 days at 25-30 ℃ and 90-95% humidity; after the culture, Ganoderma mycelium and Ganoderma fruiting body are obtained, and are pulverized to particle size of below 10 μm to obtain culture.
6. The method according to claim 1, wherein in the step (3), the mass ratio of the culture to the deionized water to the 1, 3-propanediol is 1: 5-7: 0.06-0.08.
7. The method according to claim 1, wherein in the step (3), the first standing time is 2 to 4 hours, and the second standing time is 5 to 8 hours.
8. The preparation method according to claim 1, wherein in the step (4), the mass ratio of the gordon euryale seed extract, the deionized water and the high-activity yeast liquid is 1: 8-10: 0.05 to 0.08.
9. The method according to claim 1, wherein in the step (4), the fermentation process conditions are as follows: fermenting for 3-4 days at 18-25 ℃.
10. A gorgon euryale seed extract-based health wine for reducing blood sugar and inhibiting cancer cell proliferation is obtained by the preparation method of claims 1-9.
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