A type of rice wine and its preparation method

Abstract

This invention discloses a type of rice wine and its preparation method in the field of rice wine brewing; the rice wine is composed of millet, water, Aspergillus oryzae, brewing yeast, and a peony seed meal extract complex; the peony seed meal extract complex is composed of a peony seed meal flavonoid phospholipid complex and a peony seed meal polypeptide-selenium complex; after the peony seed meal flavonoids and lecithin are combined, a fat-soluble complex that is easily absorbed by the intestines is formed, improving the utilization rate of peony seed meal flavonoids; inorganic selenium is converted into organic selenium through polypeptide compounding, which provides selenium element on the one hand and has a certain promoting effect on lowering cholesterol on the other hand.

Description

Technical Field

[0001] This invention belongs to the field of rice wine brewing technology, specifically referring to a type of rice wine and its preparation method. Background Technology

[0002] As people's material and cultural living standards continue to improve, consumers' demands for rice wine are also constantly increasing. The nutritional value and taste of rice wine are receiving more and more attention from consumers. As one of the oldest types of wine, rice wine has a brewing history of thousands of years. Through the co-fermentation of various microorganisms and specific fermentation processes, rice wine has unique flavor characteristics and specific functionalities. Therefore, rice wine is mellow, has a prominent aroma, is rich in nutrients, and is rich in functional factors, such as active polysaccharides, functional oligosaccharides, flavonoids, phenols, terpenes, trimethylolpropane, γ-aminobutyric acid, glutathione, etc. It has a variety of physiological functions such as anti-oxidation, anti-aging, lowering blood pressure, lowering cholesterol, improving the body's immunity, and preventing cardiovascular diseases, and is deeply loved by consumers. Summary of the Invention

[0003] In view of the above situation and to overcome the defects of the prior art, the present invention provides a rice wine and its preparation method. The present invention proposes to regulate the content of blood lipids and cholesterol in the body by adding a compound extracted from peony seed meal.

[0004] To achieve the above objectives, the technical solution adopted by the present invention is as follows:

[0005] This invention proposes a type of rice wine, which is made from the following components in parts by weight: 350-400 parts millet, 700-800 parts water, 6-7.2 parts Aspergillus oryzae, 2-3 parts brewing yeast, and 1-2 parts peony seed meal extract complex.

[0006] Preferably, the peony seed meal extract complex comprises the following components in parts by weight: 1-2 parts peony seed meal flavonoid phospholipid complex and 4-8 parts peony seed meal polypeptide-selenium complex.

[0007] Preferably, the preparation method of the peony seed meal extract complex specifically includes the following steps:

[0008] S1. After crushing the peony seed meal, pass it through a 40-50 mesh sieve to obtain peony seed meal powder. Add a 50% ethanol solution, stir evenly, seal with plastic wrap, and place in a 65-75℃ constant temperature water bath. Stir at 180-250 r / min for 95-110 min. Filter and separate the filtrate and peony seed meal residue. Continue to extract the peony seed meal residue according to step S1. Repeat the extraction 3 times to obtain the filtrate and peony seed meal residue. Store the peony seed meal residue for later use. Concentrate the filtrate and freeze-dry it to obtain the total flavonoid extract of peony seed meal.

[0009] S2. Add the total flavonoid extract of peony seed meal prepared in step S1 and lecithin to the reaction solvent anhydrous ethanol, react in a constant temperature water bath at 40-45℃ for 20-24h, remove the reaction solvent anhydrous ethanol by vacuum method, add 200-300mL of dichloromethane, stir thoroughly to dissolve, filter, collect the dichloromethane filtrate, evaporate to dryness in a water bath to obtain peony seed meal flavonoid phospholipid complex.

[0010] S3. Add the peony seed meal residue prepared in step S1 to distilled water, adjust the pH value to 8-9, put it in a 70-80℃ constant temperature water bath, stir at 180-250 r / min for 40-45 min, filter, separate the filtrate and filter cake, and continue to extract the filter cake according to step S3, repeat the extraction twice, take the filtrate, use the alkaline dissolution and acid precipitation method, adjust the pH value of the filtrate to 4-4.5, stir thoroughly, centrifuge at 4000 r / min for 5-6 min, wash the precipitate 3 times with deionized water, freeze dry to obtain peony seed meal protein powder;

[0011] S4. Add the peony seed meal protein powder prepared in step S3 to a 0.1 mol / L phosphate buffer solution, stir magnetically, adjust the pH to 8-9, add alkaline protease in a constant temperature water bath at 55-65℃, enzymatically hydrolyze for 120-140 min, then inactivate the enzyme at 95-100℃ for 15-20 min, cool to room temperature, adjust the pH to 7.0, centrifuge at 12000 r / min for 15-18 min, take the supernatant and filter it through an ultrafiltration membrane with a molecular weight cutoff of 3-10 kDa, adjust the concentration, and obtain the peony seed meal polypeptide solution.

[0012] S5. After mixing the peony seed meal polypeptide solution prepared in step S4 with a sodium selenite solution with a molar concentration of 1 mmol / L, add a cysteine ​​solution with a molar concentration of 4 mmol / L and adjust the pH value to 8.0 to obtain the peony seed meal polypeptide-selenium complex.

[0013] S6. Mix the peony seed meal flavonoid phospholipid complex prepared in step S2 and the peony seed meal polypeptide-selenium complex prepared in step S5 to obtain the peony seed meal extract complex.

[0014] Preferably, in step S1, the peony seed meal powder has a mass fraction of 3-5% in a 50% ethanol solution.

[0015] Preferably, in step S2, the mass ratio of the total flavonoid extract of peony seed meal to lecithin is 1:2-2.5;

[0016] Preferably, in step S2, the total flavonoid extract of peony seed meal has a mass concentration of 1-2 mg / mL in anhydrous ethanol;

[0017] Preferably, in step S3, the mass fraction of the peony seed meal residue in distilled water is 4-5%;

[0018] Preferably, in step S4, the mass concentration of the peony seed meal protein powder in the phosphate buffer is 30 g / L-40 g / L;

[0019] Preferably, in step S4, the concentration of the peony seed polypeptide solution is 2-3 mg / mL;

[0020] Preferably, in step S4, the amount of alkaline protease added is 4-5% of the mass of peony seed meal protein powder, and the enzyme activity is 10000U / g.

[0021] Preferably, in step S5, the volume ratio of the peony seed meal polypeptide solution, sodium selenite solution, and cysteine ​​solution is 1:3-4:3-4.

[0022] This invention also provides a method for preparing rice wine, specifically including the following steps:

[0023] (1) Soak millet in water at a constant temperature of 20℃ for 36-40h, then steam it at normal pressure for 45-55min. When the temperature drops to 30℃, add Aspergillus oryzae, stir evenly and saccharify for 60-65h, then add brewing yeast, stir evenly and pour into a fermentation tank, and ferment at a constant temperature of 30-35℃ for 4-5d to obtain the main fermentation product of rice wine.

[0024] (2) Add the peony seed meal extract complex to the main fermentation product of rice wine prepared in step (1), stir evenly and seal, age at room temperature of 18-20℃ for 15-18 days, centrifuge the lees for 15-20 minutes, take the supernatant to obtain a rice wine.

[0025] The beneficial effects achieved by this invention are as follows:

[0026] This invention adds a peony seed meal extract complex to the rice wine brewing process, increasing the nutritional components of rice wine, such as flavonoids, polypeptides, and selenium. This improves the composition of functional factors in rice wine, preserving not only the unique pure, elegant, and harmonious aroma and fresh, refreshing, and mellow taste of rice wine, but also its lipid-lowering and cholesterol-lowering effects. Flavonoids have significant free radical scavenging and antioxidant capabilities and offer some protection to the liver; however, their poor intestinal absorption leads to low utilization. The flavonoid-phospholipid complex enhances their lipid solubility, improves intestinal absorption and bioavailability, thereby improving the rice wine's overall health. Ketones lower blood lipids and cholesterol; polypeptides promote the conversion of cholesterol into bile acids, which are metabolic products of cholesterol in the liver. Polypeptides further degrade cholesterol in the liver, thereby reducing the cholesterol content in the liver; selenium is an essential trace element for the human body. Selenium supplementation is generally in the form of inorganic selenium and organic selenium. Inorganic selenium has disadvantages such as low bioavailability and high toxicity. In comparison, organic selenium has a higher absorption rate and safety. The combination of selenium and polypeptides has a certain promoting effect on lowering blood lipids and cholesterol; This invention achieves the regulation of blood lipid and cholesterol levels in the human body by adding a peony seed meal extract complex. Attached Figure Description

[0027] Figure 1 This is a diagram showing the modeling results of hyperlipidemia mice in this invention;

[0028] Figure 2 The graph shows the results of lipid-lowering and cholesterol-lowering tests performed on hyperlipidemic mice in Example 1 and Comparative Examples 1-3 of this invention.

[0029] Figure 3 The graph shows the liver function test results of hyperlipidemic mice in Example 1 and Comparative Examples 1-3 of this invention;

[0030] Figure 4 The results of liver superoxide dismutase (SOD) detection in hyperlipidemic mice in Example 1 and Comparative Examples 1-3 of this invention are shown.

[0031] The accompanying drawings are provided to further illustrate the invention and form part of the specification. They are used together with the embodiments of the invention to explain the invention and do not constitute a limitation thereof. Detailed Implementation

[0032] The technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. All other embodiments obtained by those skilled in the art based on the embodiments of the present invention without creative effort are within the scope of protection of the present invention.

[0033] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those familiar to those skilled in the art; furthermore, any methods and materials similar to or equivalent to those described herein may be applied to this invention; the preferred embodiments and materials described herein are for illustrative purposes only and do not limit the scope of this application.

[0034] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; unless otherwise specified, the experimental materials and test strains used in the following examples were purchased from commercial channels.

[0035] The reagents used in the examples were sourced from the following sources:

[0036] Ethanol (analytical grade), Cas No: 64-17-5, purchased from Nanjing Chemical Reagent Co., Ltd.

[0037] Lecithin, CasNo: 8002-43-5, Product No.: JZ00ILEY, purchased from Shanghai Kaiwei Chemical Technology Co., Ltd.;

[0038] Dichloromethane, CasNo: 75-09-2, Product No.: 40071190, purchased from China National Pharmaceutical Group Co., Ltd.;

[0039] Sodium hydroxide, CasNo: 1310-73-2, Product No.: 100197008, purchased from China National Pharmaceutical Group Co., Ltd.;

[0040] Hydrochloric acid, CasNo: 7647-01-0, Product No.: 10011008, purchased from China National Pharmaceutical Group Co., Ltd.;

[0041] Phosphate buffer, CasNo: None, Catalog No: 72013560, purchased from China National Pharmaceutical Group Co., Ltd.;

[0042] Alkaline protease, CasNo: 9014-01-1, Product No.: L-KE906, purchased from Shanghai Jiuding Chemical Technology Co., Ltd.;

[0043] Sodium selenite, CasNo: 10102-18-8, Product No.: 80118660, purchased from China National Pharmaceutical Group Co., Ltd.;

[0044] Cysteine, CasNo: 921-01-7, Product No.: D831312, purchased from Maclean's Ltd.;

[0045] Aspergillus oryzae, strain preservation number: CICC41168, originated from China Industrial Microbial Culture Collection Center;

[0046] Saccharomyces cerevisiae, strain preservation number: CICC1009, originated from China Industrial Microbial Culture Collection Center;

[0047] Example 1

[0048] This embodiment proposes a rice wine comprising the following components by weight: 350 parts millet, 700 parts water, 6 parts Aspergillus oryzae, 2 parts brewing yeast, and 1 part peony seed meal extract complex.

[0049] The preparation method of peony seed meal extract complex includes the following steps:

[0050] S1. Crush 500g of peony seed meal and pass it through a 40-mesh sieve. Add 15L of 50% ethanol solution and stir well. Seal with plastic wrap and use water bath extraction method. Place the mixture in a 65℃ constant temperature water bath and stir at 180r / min for 95min. Filter and separate the filtrate and peony seed meal residue. Continue to extract the peony seed meal residue according to step S1. Repeat the extraction 3 times. Store the peony seed meal residue for later use. Concentrate the filtrate and freeze-dry it to obtain the total flavonoid extract of peony seed meal.

[0051] S2. Take 0.5g of the total flavonoid extract of peony seed meal prepared in step S1 and 1g of lecithin, add 500mL of anhydrous ethanol as the reaction solvent, react in a constant temperature water bath at 40℃ for 20h, remove the anhydrous ethanol as the reaction solvent by vacuum method, add 200mL of dichloromethane, stir thoroughly to dissolve, filter, collect the dichloromethane filtrate, evaporate to dryness in a water bath to obtain peony seed meal flavonoid phospholipid complex;

[0052] S3. Take 50g of the peony seed meal residue prepared in step S1 and add it to 1L of distilled water. Adjust the pH value to 8 and place it in a 70℃ constant temperature water bath. Stir at 180r / min for 40min. Filter and separate the filtrate and filter cake. Continue to extract the filter cake according to step S3. Repeat the extraction twice. Take the filtrate and use the alkaline dissolution and acid precipitation method to adjust the pH value of the filtrate to 4. After stirring thoroughly, centrifuge at 4000r / min for 5min. Wash the precipitate three times with deionized water and freeze dry to obtain peony seed meal protein powder.

[0053] S4. Take 3g of the peony seed meal protein powder prepared in step S5 and add it to 100mL of 0.1mol / L phosphate buffer. Stir magnetically and adjust the pH to 8. Add 0.12g of alkaline protease with an enzyme activity of 10000U / g in a 55℃ constant temperature water bath. After enzymatic hydrolysis for 120min, inactivate the enzyme at 95℃ for 15min. Cool to room temperature, adjust the pH to 7.0, and centrifuge at 12000r / min for 15min. Take the supernatant and filter it through an ultrafiltration membrane with a molecular weight cutoff of 3kDa to obtain peony seed meal polypeptide solution.

[0054] S5. Take 10 mL of the peony seed meal polypeptide solution prepared in step S4 and mix it with 30 mL of sodium selenite solution with a molar concentration of 1 mmol / L. Then add 30 mL of cysteine ​​solution with a molar concentration of 4 mmol / L and adjust the pH value to 8.0 to obtain the peony seed meal polypeptide-selenium complex.

[0055] S6. Take 0.4g of the peony seed meal flavonoid phospholipid complex prepared in step S3 and mix it with 1.4mL of the peony seed meal polypeptide-selenium complex prepared in step S5 to obtain the peony seed meal extract complex.

[0056] This invention also provides a method for preparing rice wine, specifically including the following steps:

[0057] (1) After soaking millet in water at a constant temperature of 20℃ for 36 hours, steam it under normal pressure for 45 minutes. When the temperature drops to 30℃, add Aspergillus oryzae, stir evenly and saccharify for 60 hours. Then add brewing yeast, stir evenly and pour it into a fermentation tank. Ferment at a constant temperature of 30℃ for 4 days to obtain the main fermentation product of rice wine.

[0058] (2) Add the peony seed meal extract complex to the main fermentation product of rice wine prepared in step (1), stir evenly and seal, age at room temperature of 18℃ for 15 days, centrifuge the lees for 15 minutes, take the supernatant to obtain a rice wine.

[0059] Example 2

[0060] This embodiment proposes a rice wine comprising the following components in parts by weight: 400 parts millet, 800 parts water, 7.2 parts Aspergillus oryzae, 3 parts brewing yeast, and 2 parts peony seed meal extract complex.

[0061] The preparation method of peony seed meal extract complex includes the following steps:

[0062] S1. Crush 750g of peony seed meal and pass it through a 50-mesh sieve. Add 15L of 50% ethanol solution and stir well. Seal with plastic wrap and use water bath extraction method. Place the mixture in a 75℃ constant temperature water bath and stir at 250r / min for 110min. Filter and separate the filtrate and peony seed meal residue. Continue to extract the peony seed meal residue according to step S1. Repeat the extraction 3 times. Store the peony seed meal residue for later use. Concentrate the filtrate and freeze dry to obtain the total flavonoid extract of peony seed meal.

[0063] S2. Take 0.5g of the total flavonoid extract of peony seed meal prepared in step S1 and 1.25g of lecithin, add 250mL of anhydrous ethanol as the reaction solvent, react in a constant temperature water bath at 45℃ for 24h, remove the anhydrous ethanol as the reaction solvent by vacuum method, add 300mL of dichloromethane, stir thoroughly to dissolve, filter, collect the dichloromethane filtrate, evaporate to dryness in a water bath to obtain peony seed meal flavonoid phospholipid complex;

[0064] S3. Take 40g of the peony seed meal residue prepared in step S1 and add it to 1L of distilled water. Adjust the pH value to 9 and place it in an 80℃ constant temperature water bath. Stir at 250r / min for 45min. Filter and separate the filtrate and filter cake. Continue to extract the filter cake according to step S3. Repeat the extraction twice. Take the filtrate and use the alkaline dissolution and acid precipitation method to adjust the pH value of the filtrate to 4.5. After stirring thoroughly, centrifuge at 4000r / min for 6min. Wash the precipitate three times with deionized water and freeze dry to obtain peony seed meal protein powder.

[0065] S4. Take 4g of the peony seed meal protein powder prepared in step S5 and add it to 100mL of 0.1mol / L phosphate buffer. Stir magnetically and adjust the pH to 9. Add 0.2g of alkaline protease with an enzyme activity of 10000U / g in a 65℃ constant temperature water bath. After enzymatic hydrolysis for 140min, inactivate the enzyme at 100℃ for 20min. Cool to room temperature and adjust the pH to 7.0. Centrifuge at 12000r / min for 18min. Take the supernatant and filter it through an ultrafiltration membrane with a molecular weight cutoff of 10kDa to obtain peony seed meal polypeptide solution.

[0066] S5. Take 10 mL of the peony seed meal polypeptide solution prepared in step S4 and mix it with 40 mL of sodium selenite solution with a molar concentration of 1 mmol / L. Then add 40 mL of cysteine ​​solution with a molar concentration of 4 mmol / L and adjust the pH value to 8.0 to obtain the peony seed meal polypeptide-selenium complex.

[0067] S6. Take 0.6g of the peony seed meal flavonoid phospholipid complex prepared in step S3 and mix it with 2mL of the peony seed meal polypeptide-selenium complex prepared in step S5 to obtain the peony seed meal extract complex.

[0068] This invention also provides a method for preparing rice wine, specifically including the following steps:

[0069] (1) After soaking millet in water at a constant temperature of 20℃ for 40 hours, steam it under normal pressure for 55 minutes. When the temperature drops to 30℃, add Aspergillus oryzae, stir evenly and saccharify for 65 hours. Then add brewing yeast, stir evenly and pour it into a fermentation tank. Ferment at a constant temperature of 35℃ for 5 days to obtain the main fermentation product of rice wine.

[0070] (2) Add the peony seed meal extract complex to the main fermentation product of rice wine prepared in step (1), stir evenly and seal, age at room temperature 20℃ for 18 days, centrifuge the lees for 20 minutes, take the supernatant to obtain a rice wine.

[0071] Example 3

[0072] This embodiment proposes a rice wine comprising the following components in parts by weight: 375 parts millet, 750 parts water, 6.6 parts Aspergillus oryzae, 3.5 parts brewing yeast, and 1.5 parts peony seed meal extract complex.

[0073] The preparation method of peony seed meal extract complex includes the following steps:

[0074] S1. Crush 600g of peony seed meal and pass it through a 45-mesh sieve. Add 15L of 50% ethanol solution and stir well. Seal with plastic wrap and use water bath extraction method. Place the mixture in a 70℃ constant temperature water bath and stir at 210r / min for 100min. Filter and separate the filtrate and peony seed meal residue. Continue to extract the peony seed meal residue according to step S1. Repeat the extraction 3 times. Store the peony seed meal residue for later use. Concentrate the filtrate and freeze dry to obtain the total flavonoid extract of peony seed meal.

[0075] S2. Take 0.5g of the total flavonoid extract of peony seed meal prepared in step S1 and 1.125g of lecithin, add 330mL of anhydrous ethanol as the reaction solvent, react in a constant temperature water bath at 42℃ for 22h, remove the anhydrous ethanol as the reaction solvent by vacuum method, add 250mL of dichloromethane, stir thoroughly to dissolve, filter, collect the dichloromethane filtrate, evaporate to dryness in a water bath to obtain peony seed meal flavonoid phospholipid complex;

[0076] S3. Take 45g of the peony seed meal residue prepared in step S1 and add it to 1L of distilled water. Adjust the pH value to 8.5 and place it in a 75℃ constant temperature water bath. Stir at 210r / min for 42min. Filter and separate the filtrate and filter cake. Continue to extract the filter cake according to step S3. Repeat the extraction twice. Take the filtrate and use the alkaline dissolution and acid precipitation method to adjust the pH value of the filtrate to 4.3. After stirring thoroughly, centrifuge at 4000r / min for 5.5min. Wash the precipitate three times with deionized water and freeze dry to obtain peony seed meal protein powder.

[0077] S4. Take 3.5g of the peony seed meal protein powder prepared in step S5 and add it to 100mL of 0.1mol / L phosphate buffer. Stir magnetically and adjust the pH to 8.5. Add 0.16g of alkaline protease with an enzyme activity of 10000U / g in a 60℃ constant temperature water bath. After enzymatic hydrolysis for 130min, inactivate the enzyme at 97℃ for 17min. Cool to room temperature, adjust the pH to 7.0, and centrifuge at 12000r / min for 17min. Take the supernatant and filter it through an ultrafiltration membrane with a molecular weight cutoff of 5kDa to obtain peony seed meal polypeptide solution.

[0078] S5. Take 10 mL of the peony seed meal polypeptide solution prepared in step S4 and mix it with 35 mL of sodium selenite solution with a mass concentration of 1 mmol / L. Then add 35 mL of cysteine ​​solution with a mass concentration of 4 mmol / L and adjust the pH value to 8.0 to obtain the peony seed meal polypeptide-selenium complex.

[0079] S6. Take 0.5g of the peony seed meal flavonoid phospholipid complex prepared in step S3 and mix it with 1.8mL of the peony seed meal polypeptide-selenium complex prepared in step S5 to obtain the peony seed meal extract complex.

[0080] This invention also provides a method for preparing rice wine, specifically including the following steps:

[0081] (1) After soaking millet in water at a constant temperature of 20℃ for 38 hours, steam it under normal pressure for 50 minutes. When the temperature drops to 30℃, add Aspergillus oryzae, stir evenly and saccharify for 62 hours, then add brewing yeast, stir evenly and pour it into a fermentation tank, ferment at a constant temperature of 32℃ for 4 days to obtain the main fermentation product of rice wine.

[0082] (2) Add the peony seed meal extract complex to the main fermentation product of rice wine prepared in step (1), stir evenly and seal, age at room temperature of 19℃ for 17 days, centrifuge the lees for 17 minutes, take the supernatant to obtain a rice wine.

[0083] Comparative Example 1

[0084] This comparative example provides a rice wine and its preparation method. The only difference between this example and Example 1 is that it does not contain the peony seed meal extract complex in any of the components. The other components and their contents are the same as in Example 1.

[0085] Comparative Example 2

[0086] This comparative example provides a rice wine and its preparation method. The only difference between this example and Example 1 is that the peony seed meal extract complex does not contain the peony seed meal polypeptide-selenium complex, and the preparation method of the peony seed meal extract complex does not include step S5. The remaining components and component contents are the same as in Example 1.

[0087] Comparative Example 3

[0088] This comparative example provides a rice wine and its preparation method. The only difference between this example and Example 1 is that the peony seed meal extract complex does not contain the peony seed meal flavonoid phospholipid complex, and the preparation method of the peony seed meal extract complex does not include step S2. The remaining components and component contents are the same as in Example 1.

[0089] Establishment of a mouse model of hyperlipidemia

[0090] Hyperlipidemia is characterized by elevated levels of total cholesterol (TC), triglycerides (TG), and low-density lipoprotein (LDL) in plasma, while high-density lipoprotein (HDL) is decreased.

[0091] Male mice, weighing 24–27 g, were acclimatized for one week in an environment with a room temperature of 25±2℃, relative humidity of 50%–60%, and light and dark periods of 12 h each. They were then randomly divided into a blank control group and a model group. The blank control group was administered distilled water by gavage daily, while the model group was administered fat emulsion by gavage twice daily at a dose of 10 mL / kg each time. Behavioral responses of the mice were observed during feeding. After 30 days of continuous gavage, the mice were fasted but allowed water for 12 h. Three mice from each group were randomly selected, and blood was collected from their eyeballs. The blood was centrifuged at 3300 r / min for 15 min. The serum cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) levels were measured using an automated biochemical analyzer.

[0092] like Figure 1 The results of the hyperlipidemia mouse model showed that, compared with the blank control group, the serum TC and TG levels of the model group mice were significantly increased, LDL-C was slightly increased, and HDL-C was significantly decreased, which met the criteria for hyperlipidemia and indicated that the hyperlipidemia mouse model was successful.

[0093] Experimental Example 1

[0094] Lipid-lowering and cholesterol-lowering experiments were conducted on hyperlipidemic mice.

[0095] Experimental samples: Shaoxing wine prepared in Example 1 and Comparative Examples 1-3;

[0096] Experimental methods: Hyperlipidemic mice were randomly divided into 4 groups of 3 mice each. They were administered rice wine prepared in Example 1 and Comparative Examples 1-3 by gavage twice a day at a dose of 15 mL / kg each time. The gavage was continued for 15 days. The mice were fasted but allowed water for 12 hours. Blood was collected from the eyeballs of the mice and centrifuged at 3300 r / min for 15 min. The levels of TC, TG, HDL-C and LDL-C in the serum of the mice were measured using a fully automated biochemical analyzer.

[0097] Results analysis: such as Figure 2 The results of lipid-lowering and cholesterol-lowering tests on hyperlipidemic mice in Example 1 and Comparative Examples 1-3 show that, compared with Comparative Example 1, TC content decreased by 18.7%, TG content decreased by 34.2%, HDL-C increased by 70.5%, and LDL-C decreased by 59.3%. The peony seed meal extract complex in Example 1 has a certain alleviating effect on hyperlipidemic mice. Compared with Example 1, Comparative Example 2 showed little difference, indicating that peony seed meal flavonoids play an important role in lowering blood lipids and cholesterol. After being compounded with phospholipids, the fat solubility of flavonoids is increased, which is more conducive to the absorption of organisms and has a promoting effect. Compared with Example 1, Comparative Example 3 showed a large difference, indicating that although selenium is a trace element, it works synergistically with vitamin E on biological membranes to promote the absorption of fat-soluble substances by biological membranes and enhance the absorption of peony seed meal flavonoid phospholipid complex by organisms.

[0098] Experiment Example 2

[0099] Liver function tests were performed on hyperlipidemic mice.

[0100] Experimental samples: Shaoxing wine prepared in Example 1 and Comparative Examples 1-3;

[0101] Experimental methods: Hyperlipidemic mice were randomly divided into 4 groups of 3 mice each. They were administered rice wine prepared in Example 1 and Comparative Examples 1-3 by gavage twice a day at a dose of 15 mL / kg each time. The gavage was continued for 15 days. The mice were fasted but allowed water for 12 hours. Blood was collected from the abdominal artery of the mice and centrifuged at 3300 r / min for 15 min. The levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the mouse serum were measured.

[0102] ALT and AST are two liver enzymes that are widely present in liver cells. When liver cells are damaged, these two enzymes leak out of the cells, leading to an increase in their concentration in the blood. Therefore, the levels of these two enzymes are often used to assess whether liver function is normal.

[0103] Results analysis: such as Figure 3The liver function tests of mice with hyperlipidemia were performed in Example 1 and Comparative Examples 1-3. Compared with Comparative Examples 1-3, the AST and ALT contents of Comparative Example 1 were higher than those of Example 1 and Comparative Examples 2-3, indicating that the alcohol in rice wine caused some damage to the liver of mice with hyperlipidemia. Compared with Comparative Example 1, the AST content of Example 1 decreased by 6.7% and the ALT content decreased by 11.7%, indicating that the peony seed meal extract complex in rice wine has a protective effect on the liver.

[0104] Experimental Example 3

[0105] Experiment on liver superoxide dismutase (SOD) detection in hyperlipidemic mice

[0106] Experimental samples: Shaoxing wine prepared in Example 1 and Comparative Examples 1-3;

[0107] Experimental methods: Hyperlipidemic mice were randomly divided into 4 groups of 3 mice each. They were administered the rice wine prepared in Example 1 and Comparative Examples 1-3 by gavage twice a day at a dose of 15 mL / kg each time. The gavage was continued for 15 days. The mice were fasted but allowed water for 12 hours. The mice were then dissected. The livers were cut off at 4°C, rinsed with pre-cooled physiological saline, and the blood was absorbed by filter paper. 0.1 g of the liver was accurately weighed, added to 0.9 mL of physiological saline, homogenized, and centrifuged at 2500 r / min for 15 min. The supernatant was used for liver SOD testing.

[0108] Impaired cholesterol transport in the liver can lead to an increase in free radicals, which in turn damages the antioxidant system and causes oxidative damage. Superoxide dismutase (SOD) is an important free radical scavenging enzyme, and if SOD activity decreases, the ability to scavenge free radicals will also be weakened.

[0109] Results analysis: such as Figure 4 The results of superoxide dismutase (SOD) detection in the liver of hyperlipidemic mice in Examples 1 and Comparative Examples 1-3 showed that Comparative Example 1 had a significant difference compared with Examples 1 and Comparative Examples 2-3, indicating that the hyperlipidemia of Comparative Example 1 reduced its SOD activity, weakened its free radical scavenging ability, and decreased its antioxidant capacity. In contrast, Example 1 showed a significant difference compared with Comparative Example 1, indicating that the peony seed meal extract complex can enhance the activity of SOD in the liver and promote and enhance the liver's ability to transport cholesterol.

[0110] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

[0111] The present invention and its embodiments have been described above. This description is not restrictive, and the accompanying drawings are only one embodiment of the present invention. The actual application is not limited to this. In short, if a person skilled in the art is inspired by this description and designs a similar method and embodiment without departing from the spirit of the present invention, such design should fall within the protection scope of the present invention.

Claims

1. A method for preparing rice wine, characterized in that: The rice wine is made from the following components in parts by weight: 350-400 parts millet, 700-800 parts water, 6-7.2 parts Aspergillus oryzae, 2-3 parts brewing yeast, and 1-2 parts peony seed meal extract complex; the peony seed meal extract complex comprises the following components in parts by weight: 1-2 parts peony seed meal flavonoid phospholipid complex and 4-8 parts peony seed meal polypeptide-selenium complex. The preparation method of the rice wine specifically includes the following steps: (1) Soak millet in water at a constant temperature of 20℃ for 36-40h, then steam it at normal pressure for 45-55min. When the temperature drops to 30℃, add Aspergillus oryzae, stir evenly and saccharify for 60-65h, then add brewing yeast, stir evenly and pour into a fermentation tank, and ferment at a constant temperature of 30-35℃ for 4-5d to obtain the main fermentation product of rice wine. (2) Add the peony seed meal extract complex to the main fermentation product of rice wine prepared in step (1), stir evenly and seal, age at room temperature of 18-20℃ for 15-18 days, centrifuge the lees for 15-20 minutes, take the supernatant to obtain a rice wine. The preparation method of the peony seed meal extract complex specifically includes the following steps: S1. After crushing the peony seed meal, pass it through a 40-50 mesh sieve to obtain peony seed meal powder. Add a 50% ethanol solution, stir evenly, seal with plastic wrap, and place in a 65-75℃ constant temperature water bath. Stir at 180-250 r / min for 95-110 min. Filter and separate the filtrate and peony seed meal residue. Continue to extract the peony seed meal residue according to step S1. Repeat the extraction 3 times to obtain the filtrate and peony seed meal residue. Store the peony seed meal residue for later use. Concentrate the filtrate and freeze-dry it to obtain the total flavonoid extract of peony seed meal. S2. According to the mass ratio of total flavonoid extract of peony seed meal to lecithin of 1:2-2.5, the total flavonoid extract of peony seed meal prepared in step S1 and lecithin are added to the reaction solvent anhydrous ethanol and reacted in a constant temperature water bath at 40-45℃ for 20-24h. After removing the reaction solvent anhydrous ethanol by vacuum method, 200-300mL of dichloromethane is added, and after stirring and dissolving thoroughly, the mixture is filtered, and the dichloromethane filtrate is collected and evaporated to dryness in a water bath to obtain the peony seed meal flavonoid phospholipid complex. S3. Add the peony seed meal residue prepared in step S1 to distilled water, adjust the pH value to 8-9, put it in a 70-80℃ constant temperature water bath, stir at 180-250 r / min for 40-45 min, filter, separate the filtrate and filter cake, and continue to extract the filter cake according to step S3, repeat the extraction twice, take the filtrate, use the alkaline dissolution and acid precipitation method, adjust the pH value of the filtrate to 4-4.5, stir thoroughly, centrifuge at 4000 r / min for 5-6 min, wash the precipitate 3 times with deionized water, freeze dry to obtain peony seed meal protein powder; S4. Add the peony seed meal protein powder prepared in step S3 to a 0.1 mol / L phosphate buffer solution, stir magnetically, adjust the pH to 8-9, add alkaline protease in a 55-65℃ constant temperature water bath, enzymatically hydrolyze for 120-140 min, then inactivate the enzyme at 95-100℃ for 15-20 min, cool to room temperature, adjust the pH to 7.0, centrifuge at 12000 r / min for 15-18 min, take the supernatant and filter it through an ultrafiltration membrane with a molecular weight cutoff of 3-10 kDa, adjust the concentration to obtain peony seed meal polypeptide solution; the polypeptide concentration in the peony seed meal polypeptide solution is 2-3 mg / mL. S5. According to the volume ratio of peony seed meal polypeptide solution, sodium selenite solution and cysteine ​​solution of 1:3-4:3-4, the peony seed meal polypeptide solution prepared in step S4 is mixed with sodium selenite solution with a molar concentration of 1 mmol / L, and then cysteine ​​solution with a molar concentration of 4 mmol / L is added to adjust the pH value to 8.0 to obtain peony seed meal polypeptide-selenium complex. S6. Mix the peony seed meal flavonoid phospholipid complex prepared in step S2 and the peony seed meal polypeptide-selenium complex prepared in step S5 to obtain the peony seed meal extract complex.

2. The method for preparing rice wine according to claim 1, characterized in that: In step S1, the peony seed meal powder has a mass fraction of 3-5% in a 50% ethanol solution.

3. The method for preparing rice wine according to claim 2, characterized in that: In step S2, the total flavonoid extract of peony seed meal has a mass concentration of 1-2 mg / mL in anhydrous ethanol.

4. The method for preparing rice wine according to claim 3, characterized in that: In step S3, the mass fraction of the peony seed meal residue in distilled water is 4-5%.

5. The method for preparing rice wine according to claim 4, characterized in that: In step S4, the mass concentration of the peony seed meal protein powder in the phosphate buffer is 30 g / L-40 g / L.

6. The method for preparing rice wine according to claim 5, characterized in that: In step S4, the amount of alkaline protease added is 4-5% of the mass of peony seed meal protein powder, and the enzyme activity is 10000U / g.

7. A type of rice wine, characterized in that, Prepared by the preparation method according to any one of claims 1-6.

Images