Buckwheat health wine with antioxidant properties and preparation method thereof

Abstract

The invention discloses a bitter buckwheat health wine with antioxidant properties and a preparation method thereof, and belongs to the technical field of liquor production. The preparation method of bitter buckwheat health wine comprises the following steps: using bitter buckwheat grains as raw materials, washing, soaking, draining and crushing the bitter buckwheat grains, extracting the bitter buckwheat grains once by percolation extraction to obtain bitter buckwheat extract and raw material residues; using the expanded and domesticated honeysuckle mycelium to perform secondary fermentation and secondary percolation extraction on the raw material residues, and the three extracts are respectively concentrated and vacuum freeze-dried and then mixed to obtain bitter buckwheat dry matter. Secondly, sorghum and corn are used as raw materials, and distiller's yeast is added for saccharification and fermentation to obtain base wine. Finally, the bitter buckwheat mixed dry matter is compounded with the base wine to obtain bitter buckwheat health wine with antioxidant properties. The bitter buckwheat wine obtained by the invention has antioxidant capacity, has both the nutritional components of honeysuckle and the unique flavor of bitter buckwheat, and has excellent taste.

Description

Technical Field

[0001] The invention belongs to the technical field of liquor production, and particularly relates to a tartary buckwheat health-preserving liquor with antioxidant properties and a preparation method thereof. Background Art

[0002] In the thousands of years of inheritance, liquor is not only an indispensable commodity, but also a symbol of Chinese culture. It has become a necessary special drink in important activities such as socializing and banquets. In addition, traditional medicine and modern medicine believe that liquor is sweet, spicy, and hot in nature. It has the effects of unblocking blood vessels, removing wind and cold, and strengthening the spleen and stomach. Proper drinking can speed up metabolism, enhance resistance, lower blood pressure, and prevent arteriosclerosis. There are many kinds of Chinese liquor. The starch, protein, fat, and tannin in the raw materials are closely related to the brewing performance and flavor of liquor. Tartary buckwheat contains rich protein, starch, and active ingredients that are beneficial to the human body, and can be used as an ideal raw material for brewing. Most of the bitter buckwheat wine is directly fermented and steamed by bitter buckwheat. The production of bitter buckwheat liquor is still in its early stages, mainly reflected in: ① The active ingredients of bitter buckwheat are easily lost during the processing, and the utilization rate is low, resulting in the waste of bitter buckwheat raw materials; ② The research on the preservation method of nutrients and active ingredients in bitter buckwheat is insufficient; ③ The effective ingredients in the wine are single, the content of active ingredients is small, the taste is poor, and the popularity and per capita consumption of bitter buckwheat wine are insufficient.

[0003] Tartary buckwheat is a pseudo-cereal crop with both nutritional and active ingredients. It has an ancient history of cultivation and has been identified as having the same origin as medicine and food. It is an ideal raw material for health products and food. Experimental identification shows that it is rich in nutrients such as protein, starch, dietary fiber and trace elements, and has biologically active substances that are lacking in other cereals, such as flavonoids (among which rutin and quercetin are the most studied, with a rutin content of about 0.8-3.0%), phenolic acids, bio-sugar alcohols, etc. Experimental studies have found that tartary buckwheat flavonoids have a variety of pharmacological activities and physiological functions, such as lowering blood sugar and blood lipids, enhancing the body's immunity, anti-inflammatory, anti-oxidation, anti-aging, anti-tumor, etc., which have been verified through corresponding cell experiments, mouse experiments, and in vitro transformation experiments.

[0004] Phylloporia ribis (Lonicera japonica Thunb.) (also known as honeysuckle fungus) is a medicinal and edible fungus of the genus Phylloporia in the family Pucciniaceae. It often parasitizes on honeysuckle vine plants with a long growth period. It was found that the fungus contains high-quality mycelial protein and rich amino acids. The active ingredients identified are sterols, terpenes, organic acids, polysaccharides, etc. Relevant experimental studies have found that the fungus has antioxidant, antibacterial, anti-tumor, and immunomodulatory effects. It has a long history of folk medicine and is often used to treat pharyngitis, laryngitis, tonsils, etc., with significant effects. In addition, the Ministry of Health issued an announcement approving the fermented mycelium of Phylloporia ribis as a new resource food, so the fungus has great development potential. At present, there are few reports on the composition research and other application research of the fungus. Summary of the invention

[0005] In view of the above problems, the present invention provides a tartary buckwheat health-preserving wine with antioxidant properties and a preparation method thereof. The tartary buckwheat grains are fermented with expanded honeysuckle fungi, and the active ingredients in the tartary buckwheat are fully extracted through three extractions and two fermentations to prepare a tartary buckwheat health-preserving wine with the unique flavor of honeysuckle fungi and tartary buckwheat and antioxidant properties.

[0006] The technical solution adopted by the present invention to solve the technical problem is: a method for preparing buckwheat health wine with antioxidant properties, comprising the following steps:

[0007] A. Using buckwheat seeds as raw materials, washing, soaking, draining and then crushing, and extracting by percolation extraction method to obtain buckwheat extract 1 and raw material residue 1;

[0008] B. Drying, soaking and sterilizing the raw material residue 1 obtained in step A, adding the honeysuckle fungus after propagation culture to carry out a fermentation treatment, and after the fermentation is completed, drying and crushing the mixed fermentation product, and extracting by percolation extraction method to obtain bitter buckwheat extract 2 and raw material residue 2;

[0009] C. Dry, soak and sterilize the raw material residue 2 obtained in step B, add the honeysuckle fungus after propagation culture to carry out secondary fermentation, and after the fermentation is completed, dry and crush the mixed fermentation product, and extract the bitter buckwheat extract 3 by percolation extraction method;

[0010] D. filtering, concentrating and vacuum freeze-drying the tartary buckwheat extract 1 obtained in step A, the tartary buckwheat extract 2 obtained in step B and the tartary buckwheat extract 3 obtained in step C, respectively, to obtain tartary buckwheat dry matter 1, 2 and 3, respectively;

[0011] E. The tartary buckwheat dry matter 1, 2, and 3 obtained in step D are evenly mixed and compounded with the base wine at a ratio of 0.5-1 g of the tartary buckwheat dry matter mixture / 1000 mL of the base wine to obtain a tartary buckwheat health wine with antioxidant properties.

[0012] Wherein, in the method for preparing the above-mentioned buckwheat health wine with antioxidant properties, in step E, the base wine is prepared by the following method:

[0013] a1. Using sorghum and corn as raw materials, after removing impurities, washing, crushing, mixing evenly, steaming and cooking to obtain clinker;

[0014] a2. After the clinker is spread out and cooled, add the special distiller's yeast, mix it with the clinker thoroughly, and then put it into the tank for fermentation;

[0015] a3. After fermentation, drain the yellow water to obtain the raw wine, distill the raw wine, and after distillation, clarify, store and blend to obtain the base wine.

[0016] Among them, in the method for preparing the above-mentioned buckwheat health wine with antioxidant properties, when preparing the base wine, in step a2, the special wine yeast is prepared by the following method:

[0017] b1. Using barley and rice as raw materials, soaking, drying and crushing to obtain raw material powder;

[0018] b2. Use 2 / 3-4 / 5 of the raw material powder as fermented grains, and the remaining raw material powder as coating powder, add Luzhou Laojiao Daqu (see: Li Dahe, Li Guohong. Traditional operation method of Luzhou Laojiao Daqu liquor - congratulations on the 90th anniversary of "Luzhou Laojiao Daqu liquor" winning the international gold medal at the Panama Fair [J]. Brewing, 2005, (03): 107-114.), water and the separated mold, and mix well;

[0019] b3. Incubate the mixed ingredients at a temperature of 28-35°C for 2-3 days, and then at a temperature of 35-40°C for another 4-5 days until the koji matures. Oven or sun-dry the mature koji to obtain a special distiller's koji.

[0020] Wherein, in the method for preparing the above-mentioned buckwheat health wine with antioxidant properties, when preparing the base wine, in step a1, the mass ratio of sorghum to corn is 700-1200:200-300.

[0021] Wherein, in the method for preparing the above-mentioned buckwheat health wine with antioxidant properties, when preparing the base wine, in step a2, the mass ratio of the steamed sorghum, corn and special wine yeast is 700-1200:200-300:100-200.

[0022] Wherein, in the method for preparing the above-mentioned buckwheat health wine with antioxidant properties, when preparing the base wine, in step a2, the fermentation temperature is 25-28° C., and the fermentation time is 45-60 days.

[0023] Wherein, in the method for preparing the above-mentioned buckwheat health wine with antioxidant properties, when preparing the base wine, in step a3, the alcohol content of the base wine is 55-67°.

[0024] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, when preparing the special wine yeast, in step b1, the mass ratio of barley and rice is 5-10:3-10, the soaking time is 4-8h, and the particle size of the obtained raw material powder is passed through a 100-150 mesh sieve.

[0025] Among them, in the preparation method of the above-mentioned bitter buckwheat health wine with antioxidant properties, when preparing the special wine yeast, in step b2, the amount of Luzhou Laojiao wine yeast added is 8-15% of the mass of the wine yeast, the amount of water added is 50-70% of the mass of the wine yeast; the amount of mold (which is separated from the conventional commercial wine yeast in the field) added is 5-10% of the mass of the wine yeast; the concentration of the mold is 10 7 CFU / mL.

[0026] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, in step A, the tartary buckwheat seeds with a high flavonoid content are selected as raw materials, and the flavonoid content is 2-4%.

[0027] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, in step A, the soaking time is 9-11 hours, and the crushing mesh size is 25-50 meshes.

[0028] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, in step A, the operation of the percolation extraction method is: mixing the dried and crushed mixed fermentation product with 10-20 times the mass of ethanol aqueous solution (generally 55-65% ethanol aqueous solution), soaking it at room temperature for 10-15 hours, and then placing the entire system in a percolation device for percolation extraction to obtain extract 1.

[0029] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, in step B and step C, the dried raw material residue 1 and raw material residue 2 are moistened and soaked in pure water for 1 hour and then packaged into fermentation bottles, and sterilized in a high-pressure steam autoclave, and the sterilization conditions are: 100°C, 15min.

[0030] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, in step B and step C, the expanded and domesticated honeysuckle mycelium is prepared by the following method: the culture base with honeysuckle fungi is expanded and cultured on a corn matrix (corn kernels of uniform size are selected, soaked and cleaned, sterilized by high temperature and high pressure, and packaged in a fermentation bottle), the culture temperature is 24-26°C, the culture time is 25-30 days, so that the corn matrix is ​​full of honeysuckle fungi, and the domesticated honeysuckle mycelium after expansion and domestication is obtained.

[0031] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, in step B and step C, the ratio of the added amount of the expanded and domesticated honeysuckle mycelium to the raw material residues 1 and 2 obtained in step A and step B is 20-25 corn kernels: 300-500g raw material residue.

[0032] In the present invention, since the mycelia of the honeysuckle fungus grow on the corn substrate (ie corn kernels), the present invention directly adds the corn kernels covered with mycelia.

[0033] Wherein, in the method for preparing the above-mentioned buckwheat health wine with antioxidant properties, in step B and step C, the temperature of the primary fermentation and the secondary fermentation treatment is 24-26° C., and the fermentation time is 15-25 days.

[0034] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, in step B and step C, the operation of the percolation extraction method is: mixing the dried and crushed mixed fermentation product with 10-30 times the mass of ethanol aqueous solution (using 60-65% ethanol aqueous solution), soaking it at room temperature for 15-20 hours, and then placing the entire system in a percolation device for percolation extraction to obtain extract 2 and extract 3.

[0035] Among them, in the preparation method of the above-mentioned buckwheat health wine with antioxidant properties, in step D, the filtration conditions are: vacuum filtration using a 0.22μm filter membrane; the concentration conditions are: vacuum rotary evaporation at a temperature of 45-50°C and a rotation speed of 90-100r / min (the purpose is to remove most of the ethanol and water, generally making the ratio of the final remaining volume to the original volume be 1:10-15).

[0036] Wherein, in the method for preparing the above-mentioned buckwheat health wine with antioxidant properties, in step D, the vacuum freeze-drying is: vacuum freeze-drying at -77 to -80°C for 48 to 60 hours.

[0037] A tartary buckwheat health-preserving wine with antioxidant properties is prepared by the preparation method of the tartary buckwheat health-preserving wine with antioxidant properties.

[0038] Beneficial effects of the present invention: The buckwheat health-preserving wine provided by the present invention selects buckwheat grains rich in nutrients and effective ingredients as raw materials, and effectively integrates the healthy ingredients of honeysuckle fungus to enhance the antioxidant capacity. The obtained buckwheat wine has both the nutrients of honeysuckle fungus and the unique flavor of buckwheat, and has excellent taste.

[0039] The present invention adopts a percolation extraction method to extract the active ingredients in bitter buckwheat. Compared with other extraction methods, the method has the characteristics of simple operation, high extraction rate, protection of active ingredients, etc. After three extractions, extracts 1, 2, and 3 are obtained, and then mixed, the effective ingredients in bitter buckwheat can be fully extracted, the utilization rate of bitter buckwheat raw materials is improved, and the bitter buckwheat can be combined with other nutrients through the fermentation increase of honeysuckle fungi, and the partially extracted components such as polyphenols and flavonoids are released, so as to achieve further transformation and value-added of the active ingredients of bitter buckwheat. Secondly, the present invention uses honeysuckle fungi to perform secondary fermentation on the bitter buckwheat powder (raw material residue 1) after the primary extraction. During the fermentation process, not only the transformation and increase of the bitter buckwheat grain components are achieved, but also more active ingredients and flavor components in honeysuckle fungi can be incorporated, giving full play to the solid-state fermentation advantages of honeysuckle fungi, and improving the health value of the health-preserving wine finally obtained, thereby expanding the next application path of honeysuckle fungi, fungus fermentation technology, and bitter buckwheat.

[0040] The present invention adopts a base wine compounding method to prepare bitter buckwheat health wine, and the obtained bitter buckwheat health wine is rich in multiple elements beneficial to health, can be absorbed by the human body, and has a realistic anti-oxidation effect. The present invention improves the utilization rate of bitter buckwheat grains through multiple fermentations, greatly improves the output of bitter buckwheat health wine, reduces production costs, and provides production space and profit space for industrial production; and effectively incorporates the healthy ingredients of honeysuckle fungus to improve antioxidant capacity, becoming a new type of health wine product that is different from traditional bitter buckwheat wine in terms of product aspects, cross-border integration and healthier, and provides a unique idea for the development of bitter buckwheat health wine. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 is the standard curve of total flavonoids (in terms of rutin) in the embodiment of the present invention;

[0042] Figure 2 is the standard curve of polyphenols (in terms of gallic acid) in the embodiments of the present invention;

[0043] Figure 3 The standard curve of rutin in the embodiment of the present invention;

[0044] Figure 4 The triterpene standard curve in the embodiment of the present invention;

[0045] Figure 5 The VC-ABTS standard working curve in the embodiment of the present invention;

[0046] Figure 6 It is the standard working curve of VC-DPPH in the embodiment of the present invention. DETAILED DESCRIPTION

[0047] The technical solution of the present invention can be specifically implemented in the following manner.

[0048] The method for preparing tartary buckwheat health wine with antioxidant properties comprises the following steps:

[0049] A. Using buckwheat seeds as raw materials, washing, soaking and then crushing, and extracting by percolation extraction method to obtain buckwheat extract 1 and raw material residue 1;

[0050] B. Dry, soak and sterilize the raw material residue 1 obtained in step A, and then add the honeysuckle fungus after propagation culture to carry out a mixed fermentation process. After the fermentation is completed, dry and crush the mixed fermentation product, and use the percolation extraction method to obtain the bitter buckwheat extract 2 and the raw material residue 2;

[0051] C. Dry, soak and sterilize the raw material residue 2 obtained in step B, add the honeysuckle fungus after propagation culture to carry out secondary mixed fermentation treatment, and after the fermentation is completed, dry and crush the mixed fermentation product, and extract the bitter buckwheat extract 3 by percolation extraction method;

[0052] D. filtering, concentrating and vacuum freeze-drying the tartary buckwheat extract 1 obtained in step A, the tartary buckwheat extract 2 obtained in step B and the tartary buckwheat extract 3 obtained in step C, respectively, to obtain tartary buckwheat dry matter 1, 2 and 3, respectively;

[0053] E. Evenly mix the tartary buckwheat dry matter 1, 2, and 3 obtained in step D and compound them with the base wine, and mix them at a ratio of 0.5 g to 1 g of the tartary buckwheat dry matter mixture / 1000 mL of the base wine to obtain a tartary buckwheat health wine with antioxidant properties;

[0054] In step E of the present invention, the base wine is prepared by the following method:

[0055] a1. Using sorghum and corn as raw materials, after removing impurities, washing, crushing, mixing evenly, steaming and cooking to obtain clinker;

[0056] a2. After the clinker is spread out and cooled, add the special distiller's yeast, mix it with the clinker thoroughly, and then put it into the tank for fermentation;

[0057] a3. After fermentation, drain the yellow water to obtain the raw wine, distill the raw wine, and after distillation, clarify, store and blend to obtain the base wine.

[0058] Wherein, the special distiller's yeast described in the base wine preparation method is prepared by the following method:

[0059] b1. Using barley and rice as raw materials, soaking, drying and crushing to obtain raw material powder;

[0060] b2. Use 2 / 3 to 4 / 5 of the raw material powder as fermented grains, and the remaining raw material powder as coating powder, add Luzhou Laojiao Daqu (see: Li Dahe, Li Guohong. Traditional operation method of Luzhou Laojiao Daqu liquor - congratulations on the 90th anniversary of "Luzhou Laojiao Daqu liquor" winning the international gold medal at the Panama Fair [J]. Brewing, 2005, (03): 107-114.), water and the separated mold, and mix well;

[0061] b3. Incubate the mixed ingredients at a temperature of 28-35°C for 2-3 days, and then at a temperature of 35-40°C for another 4-5 days until the koji matures. Oven or sun-dry the mature koji to obtain a special distiller's koji.

[0062] When the base wine is prepared by the present invention, in step a1, the mass ratio of sorghum to corn is 700-1200:200-300; in step a2, the mass ratio of steamed sorghum, corn and special distiller's yeast is 700-1200:200-300:100-200, the fermentation temperature is 25-28 DEG C, and the fermentation time is 45-60 days; in step a3, the alcohol content of the base wine is 55-67 degrees.

[0063] When preparing the special distiller's yeast according to the present invention, in step b1, the mass ratio of the barley to the rice is 5-10:3-10, the soaking time is 4-8 hours, and the particle size of the obtained raw material powder is sieved through a 100-150 mesh sieve; in step b2, the amount of the Luzhou Laojiao brewing koji added is 8-15% of the mass of the fermented grains, the amount of water added is 50-70% of the mass of the fermented grains; the amount of the mold (which is separated from the conventional commercial distiller's yeast in the art) added is 5-10% of the mass of the fermented grains; the concentration of the mold is 10 7 CFU / mL.

[0064] In step A of the present invention, the tartary buckwheat seeds with a high flavonoid content are selected as raw materials, and the flavonoid content is 2% to 4%.

[0065] The invention utilizes the percolation extraction method to extract bitter buckwheat more completely. The method is simple, the equipment is easy to operate and the cost is low.

[0066] In step A of the present invention, the soaking time is 9 to 11 hours, and the crushing mesh size is 25 to 50 meshes; the operation of the percolation extraction method is: the mixed fermentation product after drying and crushing is mixed with 10 to 20 times the mass of ethanol aqueous solution (generally 55 to 65% ethanol aqueous solution), soaked at room temperature for 10 to 15 hours, and then the whole system is placed in a percolation device for percolation extraction to obtain extract 1.

[0067] In the present invention, the strains used are propagated and domesticated in a corn matrix, so that the strains are adapted to the grain matrix, thereby increasing the growth and fermentation speed of the strains and the hydrolase production speed. By using the propagated and domesticated honeysuckle fungi to ferment the once-extracted tartary buckwheat grains, the hydrolase produced in the fermentation process is used to decompose the macromolecular substances in the plant material to release the active substances combined with the macromolecular substances, so that the mixed fermentation product is rich in tartary buckwheat active ingredients (such as phenolic acids and flavonoids in tartary buckwheat grains...), and the active ingredients in the strains (polysaccharides, sterol compounds, organic acids, triterpenoid compounds...) can be added at the same time.

[0068] In step B and step C of the present invention, the dried raw material residue 1 and raw material residue 2 are moistened and soaked in pure water for 1 hour and then packaged into fermentation bottles, and sterilized in a high-pressure steam autoclave under the sterilization conditions of 100° C. for 15 minutes.

[0069] In step B and step C of the present invention, the expanded and domesticated honeysuckle mycelium is prepared by the following method: the culture base with honeysuckle fungi is expanded and cultured on a corn matrix (corn kernels of uniform size are selected, soaked and cleaned, sterilized at high temperature and high pressure, and packaged in a fermentation bottle), the culture temperature is 24-26°C, the culture time is 25-30 days, so that the corn matrix is ​​full of honeysuckle fungi, and the domesticated honeysuckle fungus mycelium after expansion and domestication is obtained.

[0070] In step B and step C of the present invention, the ratio of the added amount of the expanded and domesticated honeysuckle mycelium to the raw material residues 1 and 2 obtained in step A and step B is 20 to 25 corn kernels: 300 to 500 g of raw material residues.

[0071] In the present invention, since the mycelia of the honeysuckle fungus grow on the corn substrate (ie corn kernels), the present invention directly adds the corn kernels covered with mycelia.

[0072] In step B and step C of the present invention, the temperature of the primary fermentation and the secondary fermentation is 24-26° C., and the fermentation time is 15-25 days.

[0073] In step B and step C of the present invention, the operation of the percolation extraction method is: mixing the dried and crushed mixed fermentation product with 10 to 30 times the mass of an ethanol aqueous solution (using a 60 to 65% ethanol aqueous solution), soaking it at room temperature for 15 to 20 hours, and then placing the entire system in a percolation device for percolation extraction to obtain extract 2 and extract 3.

[0074] In step D, the filtering conditions are: vacuum filtration using a 0.22 μm filter membrane; the concentration conditions are: vacuum rotary evaporation at a temperature of 45 to 50° C. and a rotation speed of 90 to 100 r / min (the purpose is to remove most of the ethanol and water, generally making the ratio of the final remaining volume to the original volume 1:10 to 15); the vacuum freeze-drying is: vacuum freeze-drying at -77 to -80° C. for 48 to 60 hours.

[0075] A tartary buckwheat health-preserving wine with antioxidant properties is prepared by the preparation method of the tartary buckwheat health-preserving wine with antioxidant properties.

[0076] The technical solutions and effects of the present invention are further described in detail below through practical examples, but the protection scope of the present invention is not limited to the scope of the embodiments.

[0077] Preparation of honeysuckle after expansion culture:

[0078] The culture base with honeysuckle fungi is propagated and cultured on a corn matrix (corn kernels of uniform size are selected, soaked and cleaned, sterilized by high temperature and high pressure, and packaged in a fermentation bottle) at a culture temperature of 24-26°C and a culture time of 25-30 days, so that the corn matrix is ​​covered with honeysuckle fungi, thereby obtaining the domesticated honeysuckle fungi hyphae after propagation and domestication.

[0079] The special distiller's yeast is prepared by the following method:

[0080] ① Rice soaking: select barley and rice (mass ratio of 5:3) as raw materials, soak for 8 hours, dry and crush the soaked raw materials, and sieve them with a 150-mesh sieve to obtain raw material powder; ② Ingredient inoculation: use 3 / 4 of the raw material powder as fermented grains, and the rest as coating powder, add 15% of the mass of Luzhou Laojiao brewing Daqu, 60% of water and 8% of mold separated from commercially available fermented grains, stir and mix; ③ Blank making and koji training: make the mixed ingredients into round koji blanks, place the koji blanks in a culture room at 35°C, culture for 3 days, then set the temperature to 40°C, continue to culture for 5 days, and the koji is mature. Take out the mature koji and dry it at 45°C for storage.

[0081] Example 1

[0082] The preparation of bitter buckwheat health wine with antioxidant effect includes the following raw materials by weight: 1000 parts of sorghum, 200 parts of corn, 500 parts of bitter buckwheat grains, 100 parts of special wine yeast, and corn matrix covered with honeysuckle fungi.

[0083] The preparation method of the health-preserving wine comprises the following steps:

[0084] 1. Select bitter buckwheat seeds with high flavonoid content, clean them, remove impurities, soak them in clean water for 10 hours, remove excess water, and grind them. Use 1.5 times the volume of 65% ethanol aqueous solution to soak the ground bitter buckwheat flour at room temperature for 20 hours, then place the whole system in a percolation device for percolation extraction, use 20 times the volume of 65% ethanol solution to extract, and extract to obtain bitter buckwheat extract 1 and raw material residue 1;

[0085] 2. Dry the raw material residue 1 to remove excess ethanol and water, soak it in pure water, drain it, sterilize it, and then add the expanded and domesticated honeysuckle mycelium (i.e., the corn matrix covered with honeysuckle mycelium) to carry out a solid-state fermentation treatment (the inoculation amount is 20 (grains) of corn / 300 (g) of raw material residue 1, and ferment it at 26°C for 25 days). After the fermentation is completed, dry and crush the mixed fermentation product, soak the crushed mixed fermentation product in 1.5 times the volume of 65% ethanol aqueous solution at room temperature for 20 hours, and then place the whole system in a percolation device for percolation extraction, and extract it with 20 times the volume of 65% ethanol solution to obtain bitter buckwheat extract 2 and raw material residue 2;

[0086] 3. Dry the raw material residue 2 to remove excess ethanol and water, soak it in pure water, drain it, sterilize it, and then add the expanded and domesticated honeysuckle fungus (i.e., the corn matrix covered with honeysuckle hyphae) to carry out secondary solid-state fermentation (the inoculation amount is 20 (grains) of corn / 300 (g) of raw material residue 2, and ferment it at 26°C for 25 days). After the fermentation is completed, dry and crush the mixed fermentation product, soak the crushed mixed fermentation product in 1.5 times the volume of 65% ethanol aqueous solution at room temperature for 20 hours, and then place the whole system in a percolation device for percolation extraction, and extract it with 20 times the volume of 65% ethanol solution to obtain extract 3 and raw material residue 3;

[0087] 4. The bitter buckwheat extract 1, bitter buckwheat extract 2 and bitter buckwheat extract 3 obtained by three extractions were respectively filtered and impurities were removed by a vacuum filtration device with a 0.22 μm filter membrane, and then most of the ethanol and water were removed by a vacuum rotary evaporator (conditions: temperature 45° C., speed 100 r / min), so that the ratio of the final remaining volume to the original volume was 1:10, and the obtained concentrated solution was placed in a -80° C. freezer for quick freezing, and then the concentrated solution was freeze-dried by a vacuum freeze-drying process (conditions: temperature of -77 to -80° C., time of 60 h) to obtain bitter buckwheat dry matter 1, 2, 3;

[0088] 5. The steps for preparing base wine are as follows:

[0089] ① Weigh sorghum and corn, remove impurities, wash, crush, mix evenly, and then steam and cook to obtain clinker;

[0090] ② After the clinker is spread out and cooled, add the special distiller's yeast (the mass ratio of sorghum, corn and special distiller's yeast is 700:200:20), mix it with the clinker thoroughly, and then put it into the tank for fermentation (fermentation temperature 28℃, fermentation time 60d);

[0091] ③After fermentation, drain the yellow water to obtain the wine material, distill the wine material, cut off the head and tail, and re-distill the tail. After distillation, clarify, store, and blend to obtain the base wine;

[0092] 6. Evenly mix the tartary buckwheat dry matter 1, 2, and 3 and compound them with the base wine at a ratio of 1 g of the tartary buckwheat dry matter mixture / 1000 mL of the base wine to obtain a tartary buckwheat health wine with antioxidant properties.

[0093] Example 2

[0094] The preparation of bitter buckwheat health wine with antioxidant effect includes the following raw materials by weight: 1200g sorghum, 300g corn, 600g bitter buckwheat grains, 200g special wine yeast, and corn matrix covered with honeysuckle fungi.

[0095] The preparation method of the health-preserving wine comprises the following steps:

[0096] 1. Select bitter buckwheat seeds with high flavonoid content, clean them, remove impurities, soak them in clean water for 15 hours, remove excess water, and grind them. The ground bitter buckwheat powder is soaked in 1.5 times the volume of 65% ethanol aqueous solution at room temperature for 20 hours, and then the whole system is placed in a percolation device for percolation extraction, and extracted with 20 times the volume of 55% ethanol solution to obtain bitter buckwheat extract 1 and raw material residue 1;

[0097] 2. Dry the raw material residue 1 to remove excess ethanol and water, soak it in pure water, drain it, sterilize it, and then add the expanded and domesticated honeysuckle mycelium (i.e., the corn matrix covered with honeysuckle mycelium) to carry out a solid-state fermentation treatment (the inoculation amount is 25 (grains) of corn / 300 (g) of raw material residue 1, and ferment it at 26°C for 25 days). After the fermentation is completed, dry and crush the mixed fermentation product, soak the crushed mixed fermentation product in 1.5 times the volume of 65% ethanol aqueous solution at room temperature for 20 hours, and then place the whole system in a percolation device for percolation extraction, and extract it with 30 times the volume of 65% ethanol solution to obtain bitter buckwheat extract 2 and raw material residue 2;

[0098] 3. Dry the raw material residue 2 to remove excess ethanol and water, soak it in pure water, drain it, sterilize it, and then add the expanded and domesticated honeysuckle fungus (i.e., the corn matrix covered with honeysuckle hyphae) for secondary solid-state fermentation (the inoculation amount is 20 (grains) of corn / 300 (g) of raw material residue 2, and ferment it at 26°C for 25 days). After the fermentation is completed, dry and crush the mixed fermentation product, soak the crushed mixed fermentation product in 1.5 times the volume of 65% ethanol aqueous solution at room temperature for 20 hours, and then place the whole system in a percolation device for percolation extraction, and extract it with 1g / 20 times the volume of 65% ethanol solution to obtain extract 3 and raw material residue 3;

[0099] 4. The bitter buckwheat extract 1, bitter buckwheat extract 2 and bitter buckwheat extract 3 obtained by three extractions were respectively filtered and impurities were removed by a vacuum filtration device with a 0.22 μm filter membrane, and then most of the ethanol and water were removed by a vacuum rotary evaporator (conditions: temperature 45° C., speed 100 r / min), so that the ratio of the final remaining volume to the original volume was 1:10, and the obtained concentrated solution was placed in a -80° C. freezer for quick freezing, and then the concentrated solution was freeze-dried by a vacuum freeze-drying process (conditions: temperature of -77 to -80° C., time of 60 h) to obtain bitter buckwheat dry matter 1, 2, 3;

[0100] 5. The steps for preparing base wine are as follows:

[0101] ① Weigh sorghum and corn, remove impurities, wash, crush, mix evenly, and then steam and cook to obtain clinker;

[0102] ② After the clinker is spread out and cooled, add the special distiller's yeast (the mass ratio of sorghum, corn and special distiller's yeast is 1200:300:200), mix it with the clinker thoroughly, and then put it into the tank for fermentation (fermentation temperature 28°C, fermentation time 60d);

[0103] ③After fermentation, drain the yellow water to obtain the wine material, distill the wine material, cut off the head and tail, and re-distill the tail. After distillation, clarify, store, and blend to obtain the base wine;

[0104] 6. Evenly mix the tartary buckwheat dry matter 1, 2, and 3 and compound them with the base wine at a ratio of 0.5 g of the tartary buckwheat dry matter mixture / 1000 mL of the base wine to obtain a tartary buckwheat health wine with antioxidant properties.

[0105] Composition and antioxidant activity testing

[0106] 1. Determination of active ingredients

[0107] The finished wines in Examples 1 and 2 were tested by chemical methods and high performance liquid chromatography. Other conditions were kept constant, and the flavonoid components, rutin, total phenols, triterpenes and other active ingredients of the soaked and extracted sorghum wine and the original wine were determined by establishing a rutin standard curve, a gallic acid standard curve, and an oleanolic acid standard curve, respectively, to obtain the experimental data in Table 1;

[0108] 1. Determination method

[0109] (1) Determination of total flavonoids

[0110] The total flavonoid content in buckwheat fermentation was determined by aluminum chloride colorimetry (see: Li Xin. Establishment of analytical method for flavonoid compounds in buckwheat [D]; Chinese Academy of Agricultural Sciences, 2010.).

[0111] Use rutin as the standard to develop a standard curve. Take 0.2mg / mL rutin standard solution (mL) and take 0, 0.25, 0.5, 1, 2, 3, 4 (mL) in a gradient into a 10mL colorimetric tube, first add 2mL aluminum chloride solution, then add 3mL potassium acetate solution, and then dilute to 5mL with 70% methanol solution. Let it react at room temperature for 30min, measure the OD value at 420nm and record it, and make a blank control at the same time. Use the net OD value (i.e. the absorbance of the working solution minus the absorbance of the blank test) as the horizontal axis and the concentration of the rutin standard solution as the vertical axis to draw a standard working curve, as shown below: Figure 1 The standard curve regression equation is Y=0.0559X-0.0025, the linear coefficient R2=0.9999, and the linear relationship is good in the range of 0.0056-0.0896 mg / mL.

[0112] In the sample determination, follow the above steps. If it exceeds the linear range, it needs to be appropriately diluted. Take 1mL of sample in a 10mL colorimetric tube, first add 2mL of aluminum chloride solution, then add 3mL of potassium acetate solution, and then dilute to 5mL with 70% methanol solution. Let it react at room temperature for 30min. Use tube 0 as a blank control, measure the OD value at 420nm and record it, and do a blank control at the same time. Calculate the total flavonoid content in the buckwheat extract according to the standard curve and the total flavonoid yield calculation formula.

[0113] (2) Determination of total phenols

[0114] The total polyphenols in buckwheat were determined by the Folin phenol colorimetric method. (See: Guo Qiancheng. Study on the extraction process of total phenols and total flavonoids from buckwheat stems [J]. China Agricultural Abstracts-Agricultural Engineering, 2020, with some modifications)

[0115] Weigh 0.500g of gallic acid, dissolve it in water and make up to 100mL, use this mother liquor to prepare a series of standard solutions with mass concentrations of 0, 4, 8, 12, 20, and 30mg / L, respectively, take 1mL of the series of standard solutions into 10mL centrifuge tubes, add 2.5mL of 10% Folin-Ciocalteu color developer and mix well for 3-8min, add 2.5mL of 15% sodium carbonate solution respectively, mix well, place in a 40℃ water bath for 1 hour, cool for 20 minutes, and measure the absorbance at 760nm. Use a UV-visible spectrophotometer to measure the absorbance value (A) at 765nm, measure three times in parallel, and draw a standard curve, such as Figure 2 shown.

[0116] (3) Determination of rutin content:

[0117] The chromatographic conditions are as follows: column: Damonsil C18 column, mobile phase: methanol-0.5% phosphoric acid aqueous solution (40:60); column temperature 40°C; flow rate 1.0 mL / min; detection wavelength 390 nm; injection volume 10 μL, detection time 5 min. Draw a standard curve, such as Figure 3 shown.

[0118] like Figure 3 As shown, the regression equation of the rutin standard curve is Y = 6.3517X-27.702, the linear coefficient R2 = 0.9994, and the linear relationship is good in the range of 0.0392-0.558 mg / mL;

[0119] (4) Triterpene content detection

[0120] The chromatographic conditions were as follows: chromatographic column: C18 (250 mm × 4.6 mm, 5 μm); mobile phase: methanol: pure water = 91:9 (pH adjusted to 3.0 with phosphoric acid), flow rate 0.6 mL / min, column temperature at room temperature, detection wavelength 210 nm, injection volume 20 μL, and detection time 25 min.

[0121] Accurately weigh 0.2mL, 0.5mL, 1mL, 2mL, and 5mL of the standard solution of triterpene standard (oleanolic acid), dilute to 10mL with methanol, and obtain a series of standard working solutions with concentrations of 2μg / mL, 5μg / mL, 10μg / mL, 20μg / mL, and 50μg / mL, respectively. After passing through a 0.22μm membrane, store at 4℃ in the dark for later use. Determine the chromatogram of the standard and the sample, and draw a standard curve, such as Figure 4 The regression equation of the triterpene standard curve is Y=23.730X+13.236, the linear coefficient R2=0.9998, and the linear relationship is good in the range of 2-50 μg / mL (r=0.9998).

[0122] 2. The active ingredient contents in the finished wines obtained in Example 1 and Example 2 were measured respectively, as shown in the following table:

[0123] Table 1 Main component detection results in Examples 1 and 2

[0124] Measurement items Total flavonoids mg / L Total phenol mg / L Rutin mg / L Triterpenes mg / L Example 1 138.8±2.02 112±0.51 112.8±1.15 105.5±1.10 Example 2 85.6±0.12 69±2.16 76.4±0.85 72.1±0.22

[0125] 2. The antioxidant activity in Examples 1 and 2 was tested. The testing method was as follows: VC equivalent, ABTS and DPPH were used as indicators for the determination of antioxidant capacity. The test results are shown in Table 2.

[0126] 1. Determination of ABTS free radical scavenging activity

[0127] (1) Preparation of Vc standard curve

[0128] Vc diluent: pipette 0.8, 1.2, 1.6, 2.0, 2.4, 2.8 mL of Vc standard solution into a 10 mL brown volumetric flask, dilute to volume with 70% methanol to prepare Vc diluents with concentrations of 4, 6, 8, 10, 12, 14 μg / mL, and refrigerate at 4°C away from light.

[0129] Take 1mL of Vc dilution (if the clearance rate is too high, dilute it), add 4mL of ABTS, shake well, let it stand in a dark place for 10 minutes, and measure the OD value at a wavelength of 734nm as A1; take another 1mL of Vc dilution, add 4mL of 70% methanol solution, shake well, let it stand in a dark place for 30 minutes, and measure the OD value at a wavelength of 734nm as A2; at the same time, do a blank test, add 4mL of ABTS to 1mL of 70% methanol solution, shake well, let it stand in a dark place for 30 minutes, and measure the OD value at a wavelength of 734nm as A0.

[0130]

[0131] ABTS scavenging rate ([A0-(A1-A2)]) / A0: With Vc concentration (μg / mL) as the horizontal axis and ABTS free radical scavenging rate as the vertical axis, a standard working curve was drawn. The standard working curve is shown in Figure 2. Figure 5 The regression equation is Y=3.23X+8.0333, the linear coefficient R2=0.9996, and the linear relationship is good.

[0132] (2) Sample determination

[0133] Take 1mL of sample solution respectively (dilute if the clearance rate is too high), add the standard solution as above, and do a blank test at the same time: add 4mL ABTS to 1mL 70% methanol solution, shake well, and let it stand in a dark place for 30min. Measure the OD value A0 at a wavelength of 734nm, calculate the ABTS clearance rate in the sample according to the formula, substitute it into the standard curve, and express the antioxidant activity in the sample as Vc / g DW(ABTS).

[0134] 2. DPPH free radical scavenging activity assay

[0135] (1) Drawing of Vc standard curve

[0136] Take 2mL of Vc dilution, add 2mL of 0.1mg / mL DPPH, shake well, let stand in a dark place for 30min, and measure the OD value at a wavelength of 517nm as A1; take another 2mL of Vc dilution, add 2mL of 70% methanol solution, shake well, let stand in a dark place for 30min, and measure the OD value at a wavelength of 517nm as A2; at the same time, do a blank test, add 2mL of 70% methanol solution, add 2mL0.1mg / mL DPPH, shake well, let stand in a dark place for 30min, and measure the OD value at a wavelength of 517nm as A0.

[0137]

[0138] DPPH radical scavenging rate = ([A0-(A1-A2)]) / A0: Calculate the DPPH radical scavenging rate of each VC dilution according to the above formula, and draw a standard working curve with VC concentration (μg / mL) as the horizontal axis and DPPH radical scavenging rate as the vertical axis. The standard working curve is as follows: Figure 6 As shown, the regression equation is Y=3.4956X+6.6006, the linear coefficient R2=0.9992, and the linear relationship is good.

[0139] (2) Sample determination

[0140] Take 2 mL of sample solution respectively (dilute if the clearance rate is too high), follow the above standard sample operation steps, and perform a blank test at the same time. Calculate the DPPH clearance rate in the sample according to the formula, substitute it into the standard curve, and express the antioxidant activity in the sample as VC / g DW(DPPH).

[0141] 3. The measurement results are shown in Table 2.

[0142] Table 2 Determination results of antioxidant activity of finished wine in Examples 1 and 2

[0143] Measurement items ABTS(mg / L) / VCEAC DPPH(mg / L) / VCEAC Example 1 337±5.09 80.9±6.15 Example 2 384±3.12 108±5.85

[0144] As can be seen from Table 2, the finished wine prepared by the present invention is rich in a variety of active ingredients, and the antioxidant activity test results show that the wine exhibits strong antioxidant activity, and therefore has a certain regulatory effect on human health.

[0145] 3. The health-preserving wine obtained in Examples 1 and 2 was subjected to sensory analysis by 20 people, and the sensory evaluation of the health-preserving wine of the present invention was obtained, as shown in Table 3:

[0146] Table 3 Sensory analysis results of finished wine in Examples 1 and 2

[0147] project Example 1 Example 2 Color Yellow, clear and transparent Light yellow, clear and transparent aroma Rich, harmonious bouquet Tongzuo taste The taste is mellow, slightly bitter at first, sweet and refreshing, with a long aftertaste Tongzuo style Unique style of this product Tongzuo Organization Status Clear and transparent, without precipitation, impurities and suspended matter Tongzuo

Claims

1. A method for preparing a bitter buckwheat health wine having antioxidant properties, characterized in that The steps include: A. Using buckwheat seeds as raw materials, washing, soaking, draining and then crushing, and extracting by percolation extraction method to obtain buckwheat extract 1 and raw material residue 1; B. drying, soaking and sterilizing the raw material residue 1 obtained in step A, adding the honeysuckle fungus after propagation culture to carry out a fermentation treatment, and after the fermentation is completed, drying and crushing the mixed fermentation product, and extracting by percolation extraction method to obtain bitter buckwheat extract 2 and raw material residue 2; C. drying, soaking and sterilizing the raw material residue 2 obtained in step B, adding the honeysuckle fungus after propagation culture to carry out secondary fermentation, and after the fermentation is completed, drying and crushing the mixed fermentation product, and extracting the bitter buckwheat extract 3 by percolation extraction method; D. filtering, concentrating and vacuum freeze-drying the tartary buckwheat extract 1 obtained in step A, the tartary buckwheat extract 2 obtained in step B and the tartary buckwheat extract 3 obtained in step C respectively to obtain tartary buckwheat dry matter 1, 2 and 3, respectively; E. The tartary buckwheat dry matter 1, 2, and 3 obtained in step D are mixed evenly and compounded with the base wine at a ratio of 0.5-1 g of the tartary buckwheat dry matter mixture / 1000 mL of the base wine to obtain a tartary buckwheat health wine with antioxidant properties; In step E, the method for preparing the base wine comprises the following steps: a1. Sorghum and corn are used as raw materials, which are cleaned, washed, crushed, mixed and steamed to obtain clinker; a2. After the clinker is spread out and cooled, add the special distiller's yeast, mix it with the clinker thoroughly, and then put it into the tank for fermentation; a3. After the fermentation is completed, the yellow water is drained to obtain the wine material, which is distilled. After the distillation is completed, it is clarified, stored, and blended to obtain the base wine; In step a2, the method for preparing the special distiller's yeast comprises the following steps: b1. Barley and rice are used as raw materials, which are soaked, dried and crushed to obtain raw powder; b2 2 / 3 ~ 4 / 5 of the raw material powder quality as fermented grains, the remaining raw material powder used as powder coating, add Luzhou Laojiao brewing Daqu, water and separated mold, mix; b3. Incubate the mixed ingredients at a temperature of 28-35°C for 2-3 days, and then at a temperature of 35-40°C for another 4-5 days until the koji matures. The mature koji is then dried in an oven or sun-dried to obtain a special wine koji.

2. The method for preparing the tartary buckwheat health wine with antioxidant properties according to claim 1, characterized in that: At least one of the following is met: In step A, tartary buckwheat grains with a high flavonoid content are selected as raw materials, wherein the flavonoid content is 2% to 4%; In step A, the soaking time is 9 to 11 hours, and the crushing mesh size is 25 to 50 meshes; In step A, the conditions of the percolation extraction method are: soaking time is 10 to 15 hours, the concentration of ethanol aqueous solution is 55 to 65%, and the amount of ethanol aqueous solution is 10 to 30 times the mass of tartary buckwheat seeds.

3. The method for preparing the tartary buckwheat health wine with antioxidant properties according to claim 1, characterized in that: At least one of the following is met: In step B and step C, the dried raw material residue 1 and raw material residue 2 are moistened and soaked in pure water for 1 hour, and then packaged into fermentation bottles, and sterilized in a high-pressure steam autoclave under the following sterilization conditions: 100° C., 15 min; In step B and step C, the honeysuckle fungus after the propagation culture is prepared by the following method: the culture base with honeysuckle fungus is propagated in a corn matrix at a culture temperature of 24-26° C. for 25-30 days, so that the corn matrix is ​​covered with honeysuckle fungus, thereby obtaining the honeysuckle fungus after the propagation culture; the ratio of the amount of the honeysuckle fungus after the propagation culture added to the raw material residues 1 and 2 obtained in step A and step B is 20-25 corn kernels: 300-500g of raw material residue; In step B and step C, the temperature of the primary fermentation and the secondary fermentation is 24-26° C., and the fermentation time is 15-25 days; In step B and step C, the conditions of the percolation extraction method are: soaking time is 15 to 20 hours, the concentration of ethanol aqueous solution is 55 to 65%, and the amount of ethanol aqueous solution is 10 to 30 times the mass of tartary buckwheat seeds.

4. The method for preparing the tartary buckwheat health wine with antioxidant properties according to claim 1, characterized in that: At least one of the following is met: In step D, the filtering conditions are: vacuum filtration using a 0.22 μm filter membrane; In step D, the concentration conditions are: vacuum rotary evaporation at a temperature of 45 to 50° C. and a rotation speed of 90 to 100 r / min, and the volume ratio of the extract to the remaining solution after evaporation is 10 to 15:1 to 3; In step D, the vacuum freeze-drying conditions are: vacuum freeze-drying at -77 to -80°C for 48 to 60 hours.

5. The method for preparing the tartary buckwheat health wine with antioxidant properties according to claim 1, characterized in that: When preparing base wine, at least one of the following must be met: Step a1, the mass ratio of sorghum to corn is 700-1200:200-300; Step a2, the mass ratio of sorghum, corn and special distiller's yeast after steaming is 700-1200:200-300:100-200; In step a2, the fermentation temperature is 25-28°C and the fermentation time is 45-60 days; In step a3, the alcohol content of the base wine is 55-67°.

6. The method for preparing the tartary buckwheat health wine with antioxidant properties according to claim 1, characterized in that: When preparing special koji, at least one of the following conditions must be met: In step b1, the mass ratio of barley to rice is 5-10:3-8; In step b1, the soaking time is 4 to 8 hours; In step b1, the particle size of the raw material powder obtained is sieved through a 100-150 mesh sieve; In step b2, the amount of Luzhou Laojiao liquor Daqu added is 8-15% of the mass of the fermented grains; In step b2, the amount of water added is 50-70% of the mass of the fermented grains; In step b2, the amount of the mold added is 5-10% of the mass of the fermented grains; the concentration of the mold is 10 7 CFU / mL.

7. A tartary buckwheat health wine with antioxidant properties, prepared by the method for preparing the tartary buckwheat health wine with antioxidant properties according to any one of claims 1 to 6.

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