Preparation method of Pu'er tea fragrant wine

Abstract

The invention relates to the field of Pu'er tea fermentation, in particular to the field of C12G3 / 04, and more particularly relates to a preparation method of Pu'er tea fragrant wine. The Pu'er tea fragrant wine provided by the invention is a wine containing high-content fennel ketone and estragole, and has the effect of preventing intestinal tracts, spleen and stomach from being stimulated during drinking, and the preparation method comprises the following steps: pile fermentation, culture saccharification, feeding of fermented grains into a tank, and steaming of the wine in a retort. The Pu'er tea fragrant wine is developed through temperature control stacking cultivation fermentation, the wine body is transparent and bright, the Pu'er tea fragrance is strong, the style is unique, in addition, caffeine contained in a system can be effectively removed through the treatment of steaming the wine in a steamer, and the influence on human health after the Pu'er tea fragrant wine is drunk is reduced.

Description

Technical Field

[0001] The invention relates to the field of Pu'er tea fermentation, in particular to the field of C12G3 / 04, and more specifically to a method for preparing Pu'er tea aroma wine. Background Art

[0002] The types of aromatic substances contained in tea include alcohols, aldehydes, ketones, esters, lactones, acids, phenols, nitrogen-containing compounds, peroxide compounds, sulfur-containing compounds, etc. So far, about 700 kinds of tea aromatic substances have been separated and identified. The strong and hearty wine, the indifferent and long tea, the two are blended to suit the quiet interest. Therefore, the integration of the cultural and consumer attributes of tea and wine to create a Pu'er tea-flavored wine will inevitably lead to a new trend of high-grade liquor consumption in the future, and there is a broad market space. Pu'er cooked tea is made from sun-dried green tea made from Yunnan's unique large-leaf tea. It is a post-fermented tea produced by long natural aging or rapid pile fermentation process under the action of microorganisms. Compared with the raw tea raw material, all kinds of chemical substances in Pu'er tea have undergone significant changes. This is because in the production process of Pu'er tea, oxidation fermentation is a key process in modern Pu'er tea production and a key factor in the formation of Pu'er tea's unique quality. During the oxidation fermentation process of Pu'er tea, the volatile substances in the tea leaves are released and transformed, forming a unique aroma. After liquor enters the human body, it causes strong stimulation to the cardiovascular system. Caffeine in tea also has the effect of stimulating nerves. Taking the two together will increase the burden on the heart. At the same time, the acetaldehyde after alcohol metabolism is not completely decomposed. Under the action of caffeine, it will enter the kidneys prematurely, thereby damaging the kidneys and reducing kidney function. Traditional soaking, percolation and ultrasonic extraction methods have not really solved the problem of the mutual restriction between tea and wine. Therefore, there is less research investment and attention on Pu'er tea wine, and such products are rare in the market.

[0003] The prior art is disclosed in CN107435019A, which discloses a preparation method of Pu-erh tea wine. Pu-erh tea is soaked in an appropriate amount of warm water and the fished-out tea leaves are immediately cooled. After cooling to room temperature, the tea leaves are taken out and placed in a drying oven for drying to obtain Pu-erh tea leaves with caffeine removed. Then, the obtained Pu-erh tea leaves with caffeine removed are further ground and pulverized to obtain ultrafine tea powder, and the ultrafine tea powder is mixed with medicinal liquor containing traditional Chinese medicine components in a certain proportion to obtain Pu-erh tea wine. However, it is still difficult to completely remove the caffeine in Pu-erh tea by soaking in warm water, and the residual caffeine will have a certain impact on human health. The prior art is disclosed in CN115651797A, which discloses a Pu-erh tea wine. The preparation raw materials of the Pu-erh tea wine include Pu-erh tea, wolfberry, glutinous rice, sugar, etc. The preparation method includes baking the corresponding raw materials and then adding white liquor for soaking to obtain Pu-erh tea wine. The disclosed Pu-erh tea wine releases the effective substances of Pu-erh tea, making it have better efficacy, and can also shorten the preparation time of Pu-erh tea wine. However, after soaking in liquor, the Pu-erh tea is not further processed, and it is very difficult to further remove the caffeine originally contained in the Pu-erh tea, and the residual caffeine may harm physical health to a certain extent.

[0004] In addition, drinking alcohol easily causes problems such as intestinal irritation and discomfort in the spleen and stomach. At present, people lack the theory and method that can be followed on how to brew wine that reduces intestinal irritation and discomfort in the spleen and stomach. The reason is that the components of different wines are different, and the components in wine are complex. Coupled with the incomplete research on substances beneficial to human health, it is difficult for people to determine which component content needs to be controlled during the wine-making process.

[0005] Therefore, it is unknown whether it is possible to brew wine with Pu-erh tea as the raw material, which can significantly reduce caffeine and at the same time has high contents of components that help relieve intestinal irritation and discomfort in the spleen and stomach. This is also a challenging research topic. Summary of the Invention

[0006] Aiming at the defects of the prior art, the purpose of the present invention is to provide a Pu-erh tea fragrance wine and its preparation method. In the obtained Pu-erh tea fragrance wine, the caffeine content is relatively low, and it has high contents of components that help relieve intestinal irritation and discomfort in the spleen and stomach, so that the obtained wine not only has the fragrance of Pu-erh tea, but also can effectively relieve the irritation of drinking alcohol to the intestines and spleen and stomach.

[0007] The present invention provides a preparation method of Pu-erh tea fragrance wine, which at least includes the following steps:

[0008] S1. Pile fermentation: Add a certain amount of water to the crushed raw Pu-erh tea for wetting operation, then add fungi for stacking and culturing fermentation, and then add it to the cooked and cooled grains and mix well, add yeast, and mix well again;

[0009] S2, bacterial cultivation and saccharification: collecting the product obtained in step S1 and cultivating bacteria in a pile, controlling the pile temperature to cultivate bacteria and saccharify to obtain mash;

[0010] S3, putting the fermented grains into the tank: putting the fermented grains into the tank after the saccharification and fermentation;

[0011] S4, steaming wine in an upper steamer: steaming the material obtained after the treatment in step S3, collecting the distillate after the steaming to obtain new wine, and then sealing the new wine and storing it for at least one year to obtain Pu'er tea flavored wine.

[0012] Preferably, the fungus includes at least one of Aspergillus niger, Aspergillus wussa, Rhizopus niger, Rhizopus oryzae, and Rhizopus chinensis.

[0013] Further preferably, the fungi are Aspergillus niger and Rhizopus oryzae.

[0014] Preferably, the Aspergillus niger accounts for 0.02-0.2% of the mass of the Pu'er raw tea residues.

[0015] Preferably, the Rhizopus oryzae accounts for 0.02-0.5% of the mass of the Pu'er raw tea residues.

[0016] Preferably, the mass ratio of Aspergillus niger to Rhizopus oryzae is 1:(2-5).

[0017] Preferably, the food includes at least one of sorghum, rice, glutinous rice, peas, corn, wheat, barley and buckwheat.

[0018] Preferably, the fermentation time is 20-24 hours.

[0019] As shown in the embodiments of the present invention, compared with ordinary corn wine, the contents of fenzone (L-fenicone, D-fenicone) and estragole in the Pu'er tea wine obtained by the present invention are significantly higher than those in ordinary corn wine. According to the research of Cui Li et al. of Jiangsu Institute of Traditional Chinese Medicine in Chinese patent CN 116602945A, it is found that fenzone has a good effect on the treatment of constipation-type irritable bowel syndrome; according to the summary of Liang Zhongyun et al. in "Research Progress on Resources and Application of Estagnol of Biological Activity" (DOI: 10.3969 / j.issn.1006-1126.2010.01.015), estragole has the function of strengthening the stomach and promoting digestion. In addition, at the same time, the caffeine and tea polyphenols contents in the Pu'er tea wine obtained by the present invention are not much different from those in ordinary corn wine.

[0020] Therefore, the contribution of the present invention to the prior art lies in: providing a wine with a high content of fenchone and estragole, which has the effect of preventing irritation to the intestines and spleen and stomach when drinking; at the same time, the caffeine content in this wine is very low, overcoming the problem that the high caffeine content in Pu-erh tea is not suitable for making wine; in addition, this wine has a typical Pu-erh tea aroma, which can meet the drinking needs of consumers who like this flavor.

[0021] In addition, when preparing the above-mentioned Pu-erh tea aroma wine, the present invention also solves the problems of strong aroma and wine body stability during the wine-making process, which are specifically described as follows:

[0022] A kind of Aspergillus niger protease in Aspergillus niger can promote the growth of microorganisms; the growth of microorganisms requires nitrogen source and energy. In the initial stage of wine-making, this enzyme can decompose proteins into amino acids under a certain humidity, providing a better nutritional environment for the growth of microorganisms. Aspergillus niger protease can also promote the production of flavor substances in wine. The amino acids generated by the decomposition of proteins are one of the main sources of alcohol substances in wine, which can make the Pu-erh tea aroma wine of the present invention have a more mellow aroma. In addition, during the fermentation process, due to the combination of polypeptides in proteins, the wine body will become turbid and there will be trace insoluble precipitates. Some proteases in Aspergillus niger can decompose these substances, making the wine have a higher transparency and better stability; in addition, the yeast used in the present invention is Saccharomyces cerevisiae. As the fermentation time prolongs, the activity of the yeast will become lower and the utilization rate of raw materials will also decrease accordingly. Aspergillus niger can promote the reproduction and growth of yeast cells, further improving the effect of the piling fermentation; Rhizopus oryzae has the characteristics of strong stability, less koji equivalent, and high liquor yield, and its amylase activity is very strong, and it is mostly used as a culture and saccharifying fungus. The inventor found that when using the above two fungi in a certain humidity environment of raw Pu-erh tea during the piling fermentation and further limiting the mass ratio of the two, the aroma of the Pu-erh tea aroma wine can be more intense, the fermentation can be more complete and the stability of the wine body can be higher.

[0023] Preferably, the mass of the grain is 8-12 times the mass of the raw Pu-erh tea residue.

[0024] As an implementable case, the mass of the grain can be 8, 9, 10, 11, or 12 times the mass of the raw Pu-erh tea residue.

[0025] Preferably, the mass of the water is 35-45% of the mass of the raw Pu-erh tea residue.

[0026] As an implementable case, the mass of the water can be 35%, 38%, 40%, or 45% of the mass of the raw Pu-erh tea residue.

[0027] Preferably, the yeast includes at least one of Saccharomyces cerevisiae, Hansenula anomala, Pichia pastoris, and Endomycopsis fibuligera.

[0028] More preferably, the yeast is Saccharomyces cerevisiae.

[0029] More preferably, the yeast accounts for 0.01-0.5% of the mass of the Pu'er raw tea residue.

[0030] As an implementable case, the yeast can be 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, or 0.5% of the mass of the Pu'er raw tea residue.

[0031] Preferably, the pile temperature is 25-35°C.

[0032] As an implementable case, the stack temperature may be 25°C, 27°C, 30°C, 33°C, or 35°C.

[0033] Preferably, the incubation and saccharification time is 20-24 hours.

[0034] As an implementable case, the incubation and saccharification time can be 20h, 21h, 22h, 23h, or 24h.

[0035] The main purpose of the incubation saccharification process is to increase the sugar content of the wine and reduce the viscosity. By hydrolyzing the branched starch in the grain, more sugar and amino acids are produced, and then the sugar is decomposed by the enzyme and converted into alcohol, which increases the taste and health care of the fermented wine, while also ensuring the stability and storage resistance of the fermented wine to the maximum extent. In the present invention, the incubation saccharification time is strictly controlled to be 20-24 hours. If the time is less than 20 hours, incomplete sugar conversion will result, affecting the fermentation efficiency and the quality of the wine; if the fermentation time is longer than 24 hours, excessive decomposition of sugar will result, producing too much acid and other impurities, thereby affecting the taste and quality of the wine.

[0036] Preferably, the closed fermentation time is 30-45 days.

[0037] As an implementable case, the closed fermentation time can be 30, 35, 40, or 45 days.

[0038] Preferably, the time for steaming the wine in the upper steamer is 60-70 minutes, and the temperature for steaming the wine in the upper steamer is 90-110°C.

[0039] As an implementable case, the time for steaming the wine in the upper steamer can be 60min, 62min, 65min, 67min, or 70min; the temperature for steaming the wine in the upper steamer can be 90℃, 95℃, 100℃, 105℃, or 110℃.

[0040] There is a certain amount of caffeine in Pu'er raw tea, which will remain in Pu'er tea wine if no further treatment is performed. The inventors found in the actual winemaking process that after the Pu'er tea wine is sealed and fermented in a tank, the caffeine contained in the system can be effectively removed by further steaming the wine in a retort. The boiling point of caffeine is 178°C. The temperature of steaming the wine in the retort in the present invention is significantly lower than the boiling point of caffeine. Therefore, caffeine in Pu'er raw tea can be effectively removed in the process of steaming the wine in the retort. In addition, this temperature will not affect the tea polyphenols in Pu'er tea. The content of tea polyphenols is significantly affected, and more tea polyphenols can be ensured to remain in the system, so that the Pu'er tea wine prepared by the present invention has a strong tea aroma. The new wine steamed in the upper steamer is sealed in a pottery jar for storage, and an oxidation-reduction reaction occurs under natural conditions, so that certain trace elements are gradually naturally combined in the wine, and the acid, ester and alcohol in the wine are dissolved, mixed and aged, so as to achieve the effect of clear and pure, full-bodied, sweet and soft, and refreshing aftertaste. The Pu'er tea wine has the aroma of aged Pu'er tea. Drinking it in moderation can also promote health and has certain health care effects.

[0041] Beneficial effects:

[0042] (I) The present invention obtains a wine with high content of fenchone and estragole through the selection of raw materials and the design of brewing process, which has the effect of preventing the intestinal tract and spleen and stomach from being stimulated when drinking. In the present invention, the caffeine contained in the system can be effectively removed by steaming the wine in the upper steamer, so that the impact of Pu'er tea-flavored wine on human health after drinking is significantly reduced.

[0043] (ii) The present invention combines the production processes of Pu'er cooked tea with Xiaoqu fragrant liquor, adds specific fungi, and controls the temperature for stacking and fermenting the bacteria to develop a fully solid-state Pu'er tea fragrant liquor production process, which has a transparent liquor body, rich Pu'er tea aroma, and a unique style.

[0044] (3) In the present invention, the Pu'er tea flavored wine is prepared by using the residue produced in the process of Pu'er raw tea processing as a raw material. Compared with using Pu'er raw tea as a raw material, the preparation cost can be effectively reduced, and the Pu'er raw tea residue can be fermented more completely, which can further improve the utilization rate of the raw materials.

[0045] (IV) The present invention uses specific fungi, namely Aspergillus niger and Rhizopus oryzae, and the quality of the two fungi is limited. Aspergillus niger and Rhizopus oryzae play a very important role in the fermentation process of Pu'er tea. They are a kind of dominant strains, which can secrete intracellular and extracellular enzymes, such as polyphenol oxidase, cellulase and protease, to promote the hydrolysis and conversion of macromolecular substances in tea, thereby improving the taste and flavor of Pu'er tea; at the same time, Aspergillus niger can also promote the production of aromatic substances in wine, so that the wine has higher transparency and clarity and better stability, and Aspergillus niger can promote the reproduction and growth of yeast cells, further improving the effect of pile fermentation. Rhizopus oryzae has the characteristics of strong stability, small amount of koji equivalent and high wine yield. The use of the two fungi together with brewing yeast can ensure that the pile fermentation is better carried out, so that the Pu'er tea fragrant wine has higher quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] Figure 1 This is a standard curve graph for test 7.

[0047] Figure 2 It is a schematic diagram of the HPLC test results of the corn wine test samples in Table 4 of the present invention. DETAILED DESCRIPTION

[0048] Example 1

[0049] The first aspect of the present embodiment provides a method for preparing Pu'er tea flavored wine, comprising the following steps:

[0050] S1, pile fermentation: add 3.5kg water to 10kg of Pu'er raw tea residue for wet operation, then add 5g of Aspergillus niger and 10g of Rhizopus oryzae for pile culture and fermentation for 24h, the culture and fermentation temperature is 30°C, then add 100kg of corn that has been steamed and cooled, mix well, add 5g of brewer's yeast, and mix well again;

[0051] S2, bacterial cultivation and saccharification: the product obtained in step S1 is subjected to bacterial cultivation in a pile for 24 hours, the pile temperature is controlled at 32° C., and bacterial cultivation and saccharification are performed to obtain a mash;

[0052] S3, the fermented grains are put into tanks: after the saccharification is completed, the fermented grains are put into tanks and fermented in a sealed manner for 30 days;

[0053] S4, steaming wine in an upper steamer: steaming the material obtained after the treatment in step S3 in an upper steamer for 60 minutes at a temperature of 90-110°C. After the steaming is completed, the distillate is collected to obtain new wine, which is then sealed in a pottery jar and stored for 365 days to obtain Pu'er tea-flavored wine.

[0054] The Pu'er raw tea residues are local Pu'er raw tea residues from Xishuangbanna, Yunnan.

[0055] The Aspergillus niger was purchased from Jining Zechuan Biotechnology Co., Ltd.

[0056] The Rhizopus oryzae was purchased from Yiyuan Kangyuan Biotechnology Co., Ltd.

[0057] The yeast was purchased from Angel Yeast Co., Ltd.

[0058] Example 2

[0059] The first aspect of the present embodiment provides a method for preparing Pu'er tea flavored wine, comprising the following steps:

[0060] S1, pile fermentation: add 4kg of water to 10kg of Pu'er raw tea residue for wet operation, then add 3g of Aspergillus niger and 6g of Rhizopus oryzae for pile culture and fermentation for 24h, the culture and fermentation temperature is 30°C, then add 100kg of corn that has been steamed and cooled, mix well, add 3g of brewer's yeast, and mix well again;

[0061] S2, bacterial cultivation and saccharification: the product obtained in step S1 is subjected to bacterial cultivation in a pile for 40 hours, the pile temperature is controlled at 30° C., and bacterial cultivation and saccharification are performed to obtain a mash;

[0062] S3, the mash is put into the tank: after the culture and saccharification is completed, the mash is put into the tank and fermented in a closed manner for 35 days;

[0063] S4, steaming wine in an upper steamer: steaming the material obtained after the treatment in step S3 in an upper steamer for 45 minutes at a temperature of 90-110°C. After the steaming is completed, the distillate is collected to obtain new wine, which is then sealed in a pottery jar and stored for 365 days to obtain Pu'er tea-flavored wine.

[0064] The Pu'er raw tea residues are local Pu'er raw tea residues from Xishuangbanna, Yunnan.

[0065] The Aspergillus niger was purchased from Jining Zechuan Biotechnology Co., Ltd.

[0066] The Rhizopus oryzae was purchased from Yiyuan Kangyuan Biotechnology Co., Ltd.

[0067] The yeast was purchased from Angel Yeast Co., Ltd.

[0068] Example 3

[0069] The first aspect of the present embodiment provides a method for preparing Pu'er tea flavored wine, comprising the following steps:

[0070] S1. Pile fermentation: Add 10 kg of raw Pu-erh tea residues to 4 kg of water for wetting, then add 8 g of Aspergillus niger and 12 g of Rhizopus oryzae, pile up for culturing and fermenting for 54 h, the culturing and fermenting temperature is 30 °C, then add it to 100 kg of cooked and cooled corn, mix well, add 5 g of Saccharomyces cerevisiae, and mix well again;

[0071] S2. Culturing and saccharification: Pile up and culture the product obtained in step S1, the time for piling up and culturing is 24 h, control the pile temperature at 32 °C, and carry out culturing and saccharification to obtain fermented grains;

[0072] S3. Fermented grains into the tank: After the culturing and saccharification are completed, put the fermented grains into the tank and ferment tightly for 30 days;

[0073] S4. Steaming for liquor: Carry out steaming for liquor on the substance obtained after the treatment in step S3, the time for steaming for liquor is 60 min, the temperature for steaming for liquor is 90 - 110 °C, collect the distillate after the steaming for liquor is completed to obtain new liquor, and then seal the new liquor in a pottery jar for storage for 365 days to obtain Pu-erh tea fragrance liquor.

[0074] The raw Pu-erh tea residues are the raw Pu-erh tea residues from Xishuangbanna, Yunnan.

[0075] The Aspergillus niger is purchased from Jining Zechuan Biotechnology Co., Ltd.

[0076] The Rhizopus oryzae is purchased from Yiyuan Kangyuan Biotechnology Co., Ltd.

[0077] The yeast is purchased from Angel Yeast Co., Ltd.

[0078] Comparative Example 1

[0079] This comparative example provides a preparation method of Pu-erh tea fragrance liquor in the first aspect, including the following steps:

[0080] S1. Pile fermentation: Add 10 kg of raw Pu-erh tea residues to 3.5 kg of water for wetting, then add 15 g of Rhizopus oryzae, pile up for culturing and fermenting for 24 h, the culturing and fermenting temperature is 30 °C, then add it to 100 kg of cooked and cooled corn, mix well, add 5 g of Saccharomyces cerevisiae, and mix well again;

[0081] S2. Culturing and saccharification: Pile up and culture the product obtained in step S1, the time for piling up and culturing is 24 h, control the pile temperature at 32 °C, and carry out culturing and saccharification to obtain fermented grains;

[0082] S3. Fermented grains into the tank: After the culturing and saccharification are completed, put the fermented grains into the tank and ferment tightly for 30 days;

[0083] S4. Steaming for liquor production: The material obtained after the treatment in step S3 is used for steaming for liquor production. The time for steaming for liquor production is 60 min, and the temperature for steaming for liquor production is 90 - 110 °C. After the steaming for liquor production is completed, the distillate is collected to obtain new liquor, and then the new liquor is sealed in a pottery jar and stored for 30 days to obtain the Pu-erh tea aroma liquor.

[0084] The Pu-erh raw tea residue mentioned above is the Pu-erh raw tea residue from Xishuangbanna, Yunnan.

[0085] The Rhizopus oryzae mentioned above is purchased from Yiyuan Kangyuan Biotechnology Co., Ltd.

[0086] The yeast mentioned above is purchased from Angel Yeast Co., Ltd.

[0087] Comparative Example 2

[0088] In the first aspect of this comparative example, a preparation method of Pu-erh tea aroma liquor is provided, including the following steps:

[0089] S1. Pile-fermentation: 10 kg of Pu-erh raw tea residue is added with 3.5 kg of water for wetting operation, then 5 g of Aspergillus niger and 10 g of Rhizopus oryzae are added for stacking and culturing fermentation for 24 h. The temperature for culturing fermentation is 30 °C, and then it is added to 100 kg of cooked and cooled corn and mixed evenly, and 5 g of brewing yeast is added and mixed evenly again;

[0090] S2. Culturing and saccharification: The product obtained in step S1 is piled up for culturing. The time for piling up for culturing is 48 h, and the temperature of the pile is controlled at 35 °C for culturing and saccharification to obtain fermented grains;

[0091] S3. Fermented grains into the tank: After the culturing and saccharification is completed, the fermented grains are put into the tank and fermented airtight for 40 days;

[0092] S4. Steaming for liquor production: The material obtained after the treatment in step S3 is used for steaming for liquor production. The time for steaming for liquor production is 60 min, and the temperature for steaming for liquor production is 200 - 220 °C. After the steaming for liquor production is completed, the distillate is collected to obtain new liquor, and then the new liquor is sealed in a pottery jar and stored for 30 days to obtain the Pu-erh tea aroma liquor.

[0093] The Pu-erh raw tea residue mentioned above is the Pu-erh raw tea residue from Xishuangbanna, Yunnan.

[0094] The Aspergillus niger mentioned above is purchased from Jining Zechuan Biotechnology Co., Ltd.

[0095] The Rhizopus oryzae mentioned above is purchased from Yiyuan Kangyuan Biotechnology Co., Ltd.

[0096] The yeast mentioned above is purchased from Angel Yeast Co., Ltd.

[0097] Performance test

[0098] 1. Detection of caffeine content

[0099] Detection objects: Raw Pu-erh tea, Pu-erh tea-flavored wines prepared in Examples 1-3 and Comparative Examples 1-2

[0100] Detection instrument: Waters I-Class Water XEVO-TQS micro

[0101] Testing unit: Shanghai Microtek

[0102] Testing method: Determination of caffeine content by ultraviolet spectrophotometry in accordance with GB / T8312-2013

[0103] Detection content: Caffeine content, and record the corresponding results in Table 1

[0104] 2. Detection of tea polyphenol content

[0105] Detection objects: Raw Pu-erh tea, Pu-erh tea-flavored wines prepared in Examples 1-3 and Comparative Examples 1-3

[0106] Detection instrument: PerKinElmer Lambda 364

[0107] Testing unit: Shanghai Microtek

[0108] Testing method: Determination of tea polyphenol content by ferric tartrate colorimetry in accordance with GB / T8313-2008. Detection content: Tea polyphenol content, and record the corresponding results in Table 1

[0109] 3. Taste detection of Pu-erh tea-flavored wine

[0110] Detection objects: Raw Pu-erh tea soaked in warm water, Pu-erh tea-flavored wines prepared in Examples 1-3 and Comparative Examples 1-3

[0111] Detection method: Let wine tasters taste the taste of the above-mentioned tea / wine respectively, and record the results in Table 1

[0112] Table 1

[0113]

[0114] 4. Testing of Pu-erh tea-flavored wine

[0115] Submitting unit: Yunnan Junwei Bingxin Biotechnology Co., Ltd.

[0116] Solid sample: Take 0.01 g of the sample (tea leaves of the same batch as in Example 1), add 1 mL of 70% methanol (preheated at 70 °C), homogenize, extract at 70 °C for 0.5 h, centrifuge at 12000 rpm for 5 min, and take the supernatant for filtration and testing.

[0117] Liquid samples are directly tested on the machine.

[0118] 4.1 Test Conditions

[0119] Instrument: Ultimate 3000;

[0120] Chromatographic column: Hypersil GOLD 250mm * 4.6mm; 5μm;

[0121] Column temperature: 25 °C;

[0122] Sample injection volume: 10 μL

[0123] Detection wavelength: 203 nm;

[0124] Mobile phase: Acetonitrile: Water = 15:85 (V:V);

[0125] Flow rate: 0.6 mL / min;

[0126] Elution method: Isocratic elution.

[0127] 4.2 Content Calculation

[0128] 1) Standard Curve Plotting

[0129] Based on the standard product concentration and peak area, plot the standard curve, as Figure 2

[0130] 2) Calculate the content of caffeine components in the sample according to the peak area of the sample detection and the standard curve

[0131] 4.3 Detection of Tea Polyphenols

[0132] 4.3.1 Experimental Methods and Principles

[0133] Plant phenolic substances have the functions of scavenging free radicals, antioxidant and anti-aging, and have high nutritional value and healthcare functions, so they are widely used in cosmetics, food, medicine and other fields.

[0134] This kit uses the Folin-Ciocalteu method to determine the content of tea polyphenols. Under alkaline conditions, phenolic substances reduce tungstomolybdic acid to produce a blue compound, which has a characteristic absorption peak at 760 nm. Read the absorbance value at 760 nm, and then calculate the content of tea polyphenols.

[0135] 4.3.2 Reagents and Materials (Table 2)

[0136] Table 2

[0137]

[0138] 4.3.3 Instrumentation

[0139] Microplate reader, 96-well plate, tabletop centrifuge, adjustable pipette, constant temperature water bath, oven, grinder / mortar, balance, oven, 30-50 mesh sieve, absolute ethanol and distilled water

[0140] 4.3.4. Procedure

[0141] Determination of tea polyphenol (TP) content:

[0142] It is recommended to select 2 samples for pre-determination before the formal experiment to understand the situation of this batch of samples, familiarize with the experimental process, and avoid waste of experimental samples and reagents!

[0143] 1. Sample preparation:

[0144] Tissue sample:

[0145] Weigh about 0.1 g of the sample (if the moisture is sufficient, the sampling mass of the sample can be increased); or weigh about 0.03 g of the dried sample (dry the sample to constant weight, crush it, and pass through a 30-50 mesh sieve to obtain the dried sample), add 1.5 mL of the extraction solution, extract with shaking at 60 °C for 2 h (if evaporation occurs, make up the volume to 1.5 mL with the extraction solution). Centrifuge at 25 °C × 12000 rpm for 10 min, and take the supernatant for testing.

[0146] 2. Detection on the machine:

[0147] ① Preheat the microplate reader for 30 min and adjust the wavelength to 750 nm.

[0148] ② Operation table (Table 3):

[0149] Table 3

[0150]

[0151] 4.4. Result calculation:

[0152] 1. Taking the concentration of each standard solution as the x-axis and the corresponding ΔA standard as the y-axis, plot the standard curve to obtain the standard equation y = kx + b, and substitute ΔA into the equation to get x (mg / mL).

[0153] 2. Calculate according to the sample protein concentration:

[0154] Tea polyphenol content (mg / mg prot) = x × V ÷ (Cpr × V) × D = x ÷ Cpr × D

[0155] 3. Calculate according to the sample mass:

[0156] Tea polyphenol content (mg / g mass) = x × V 1 ÷ (W × V 1 ÷ V) × D = 1.5x ÷ W × D

[0157] V --- Volume of the extraction solution added, 1.5 mL;

[0158] V 1 --- Volume of the sample in the reaction, 0.01 mL;

[0159] D --- Dilution factor, 1 if not diluted;

[0160] W --- Mass of the sample, g;

[0161] Cpr --- Protein concentration of the sample, mg / mL; It is recommended to use the BCA protein content detection kit of our company. Precautions:

[0162] If the measured absorbance value exceeds the linear range of the standard absorbance value: if it is higher than the highest value, it is recommended to appropriately dilute the sample to be measured with distilled water and then measure it; if it is lower than the lowest value, it is recommended to appropriately increase the sample and then measure it, and just modify it accordingly when calculating.

[0163] 4.5. Results

[0164] See Table 4 and Table 5, and Figure 2

[0165] Table 4

[0166]

[0167] The production process of the above-mentioned corn wine is as follows:

[0168] 1. Process flow of producing corn base wine

[0169] Raw materials (corn) → Soaking → Steaming grains (steaming grains with low fire, smothering grains, steaming grains with high fire) → Spreading and cooling with koji added → Piling up in the drying yard → Culturing and saccharifying → Fermenting in the pit → Distilling → Storage

[0170] 2. Operating methods and process key points of corn base wine

[0171] 3.1 Grain selection

[0172] The corn used is the high mountain corn produced locally in Menghai (above 800 m altitude). The corn kernels are complete, plump, golden yellow, and there is no mildew or insect damage.

[0173] 3.2 Soaking

[0174] Soak the grains with hot water at about 65 - 75 °C. You can also use the hot water in the cooling pool or the water for smothering grains for soaking. The hot water should submerge the grain surface by 12 - 15 cm, and the soaking time is 2 - 3 hours.

[0175] 3.3 Steaming grains

[0176] First, steam it over low heat. After the entire grain surface in the steamer is filled with steam, steam it initially for 20 minutes, then let it sit for 2 - 3 hours. Drain the water from the sitting process, and after draining, switch to high heat and steam the grains for 2 hours, then remove them from the steamer. It is required that the cooked corn kernels are thoroughly cooked, with no uncooked core inside, not sticky to the touch, and the cracking rate of the corn husks reaches over 85%.

[0177] 3.4 Cooling and Adding Qu starter

[0178] Immediately place the cooked corn removed from the steamer on the drying floor, cool it to 60 - 50°C and add 1 / 3 of the qu starter, then cool it to 50 - 40°C and add the second 1 / 3 of the qu starter. When it cools to 40 - 35°C, add all the remaining qu starter. The amount of qu starter added is 0.6%, and stir evenly.

[0179] 3.5 Piling Up and Culturing

[0180] Heap up the fermented grains mixed with the qu starter. When the temperature of the fermented grains drops to 30°C, quickly transfer them into the vat for culturing and saccharification. After 20 - 24 hours of culturing, the temperature of the fermented grains reaching 35 - 37°C is the most suitable. When the temperature of the fermented grains is too high, take measures such as transferring to another vat to cool down to prevent souring and off - flavors. Under normal culturing conditions, after another 22 - 26 hours of culturing and saccharification, the saccharification rate can reach over 85%. At the end of saccharification, there should be an obvious qu aroma, and it tastes sweet (a slight sour taste is allowed in summer).

[0181] 3.6 Fermenting in the Vat

[0182] First, spread a layer of yeast lees at the bottom of the barrel, then put the fermented grains into the barrel. The barrel should be filled in layers and stepped on firmly (moderately) to achieve a fermentation state of "slow at first, fierce in the middle, and slow in the final rise" in the barrel. After filling the barrel, cover it with the top lees and sprinkle a little rice husk. The fermentation period depends on the season. When putting it into the barrel, sprinkle the tail liquor and mix well, which can increase the aroma and taste of the liquor.

[0183] 3.7 Distilling in the Steamer

[0184] When distilling the fermented grains in the steamer, steam them slowly into the steamer, sprinkle them lightly and evenly, sprinkle the fermented grains when there is steam, be light, loose, even, and thin, without steam leakage or steam treading, with higher edges and lower in the middle. Sprinkle the head liquor from the previous steamer on the surface of the fermented grains, cover the pot lid, and seal the gap between the lid and the steamer edge. Roast the liquor with slow fire. When receiving the liquor, cut off the heads and tails to improve the liquor quality and meet the national health standards. Since the content of fusel oil in the head and tail liquor is high, it is required that for 300 kg of grains, 0.5 - 1 kg of head liquor is received. Finally, use high heat to chase the tail liquor to wash away the acidity in the yeast lees and leave the required good yeast lees for fermentation in the vat.

[0185] 3.8 Storage

[0186] The newly distilled liquor is sealed in high-quality pottery vats with a capacity of 1000 kg and stored in a cool and dry place. Storage can not only make the liquor undergo redox reactions under natural conditions, but also enable some trace elements to gradually combine and be discarded in the liquor naturally, causing the acids, esters, and alcohols in the liquor to dissolve, mix, and mature, achieving the quality of being clear, pure, full-bodied, mellow, sweet, soft, and with a clean aftertaste.

[0187] 4. Quality Standards for Corn-based Liquor

[0188] 4.1 Sensory Indexes

[0189] Light yellow or faint rice yellow, with slight turbidity allowed, sweet and clean, rich in aroma, and long aftertaste.

[0190] 4.2 Physicochemical Indexes

[0191] Alcohol content (20 °C): 45% - 48% (% vol); Methanol / (g / 100 ml): less than or equal to 0.6; Cyanide (calculated as HCN) / (mg / L): less than or equal to 8.0.

[0192] Table 5

[0193]

[0194] Analysis and Testing of Trace Components (Table 6)

[0195] Table 6 Percentage Contents of Trace Components

[0196]

[0197]

[0198] Explanation of Trace Components:

[0199] The aroma components in tea liquor include various compounds such as esters, alcohols, acids, ketones, aldehydes, pyrazines, and sulfur-containing compounds.

[0200] Esters: Ester compounds are one of the most important aroma components in tea liquor and play a decisive role in forming the typicality of tea liquor. The main esters in tea liquor include ethyl acetate, ethyl hexanoate, ethyl butyrate, and ethyl lactate, etc.

[0201] Ester substances play a very important role in tea liquor. They are the flavor components with the largest proportion in tea liquor and have a significant impact on the aroma and taste of tea liquor. The ester substances in tea liquor mainly include ethyl acetate, ethyl hexanoate, ethyl butyrate, and ethyl lactate, etc. The content of these ester substances accounts for 30% - 70% of the total flavor substance content, and their content exceeds 90% of the total ester substance content in tea liquor.

[0202] The formation of esters mainly occurs through microbial metabolism, esterase catalysis, and chemical reactions during storage. The ester-producing microorganisms in tea wine fermentation mainly include yeasts, bacteria, and molds, which metabolize and generate esters through the interaction between different microorganisms.

[0203] Alcohols: Alcohol compounds play a role in increasing the alcohol sweetness and assisting the aroma in tea wine. They are also the precursor substances for the formation of ester compounds. Important alcohols include isoamyl alcohol, isobutyl alcohol, n-butanol, and propanol, etc.

[0204] Acids: Acid compounds are the main substances for the aroma of tea wine and are also a necessary condition for the formation of ester compounds. The main components of organic acids in tea wine include acetic acid, hexanoic acid, lactic acid, and butyric acid, etc.

[0205] Acid substances play multiple roles in tea wine and have important impacts on the flavor, taste, and overall quality of tea wine.

[0206] Taste function: Acid substances are important taste substances in tea wine, which can increase the mellow feeling and aftertaste of tea wine, making the wine body more plump and coordinated. For example, an appropriate amount of hexanoic acid can increase the "cellar aroma" of the wine body, and an appropriate amount of butyric acid can also enhance the aroma of the wine body, but excessive amounts may bring unpleasant odors.

[0207] Balancing the taste: Acid substances can reduce the spiciness and bitterness of tea wine and increase the sweetness, thus balancing the taste of tea wine. Organic acids such as acetic acid and lactic acid have a significant impact on the taste of tea wine. They affect the flavor of tea wine by interacting with other components such as ethanol and esters.

[0208] Promoting ester synthesis: Acid substances are the precursor substances for ester synthesis. Ester substances play an important role in the aroma of tea wine. For example, ester compounds such as ethyl acetate and ethyl lactate are important aroma components in tea wine, and they are usually formed by the corresponding acid substances reacting with ethanol through esterification reactions.

[0209] Influencing the volatility of aroma: The addition of acid substances can affect the volatility of aroma compounds in tea wine. For example, hexanoic acid and lactic acid can promote the release of some ester substances and enhance their volatility, while acetic acid and butyric acid may have an inhibitory effect on the volatility of aroma compounds at different concentrations.

[0210] Quality control: Acid substances at different concentrations have a significant impact on the flavor profile of tea wine. Research shows that by controlling the addition amount of acid substances, the flavor of tea wine can be adjusted to achieve an ideal taste and aroma.

[0211] Healthcare function: Some acid substances such as formic acid are considered to have the effect of treating liver diseases such as hepatitis B, indicating that acid substances may be beneficial to health.

[0212] Ketones: Ketone compounds also play an important role in tea wine, such as 3-hydroxy-2-butanone, etc.

[0213] Aldehydes: Aldehyde compounds are related to the main aroma of tea wine. The main aldehydes include acetaldehyde and acetal, etc.

[0214] Pyrazines: Pyrazine compounds are particularly important in strong-flavor tea wine, such as 2,3,5-trimethylpyrazine, etc.

[0215] Sulfur-containing compounds: The aroma contribution of sulfur-containing compounds in tea wine cannot be ignored, such as dimethyl trisulfide, etc.

[0216] Other compounds: Including phenolic compounds, etc., which can also affect the aroma of tea wine.

[0217] The role of hydrocarbons in tea wine is usually not significant. The main components of tea wine are ethanol and water, as well as trace amounts of aroma components. Hydrocarbons, especially saturated hydrocarbon compounds such as alkanes, are relatively stable in chemical properties and are not easily reactive with other substances. In tea wine, they may exist as impurities or by-products, but usually do not have a significant impact on the taste, aroma or quality of tea wine. The flavor and quality of tea wine are mainly determined by components such as ethanol, esters, alcohols, aldehydes, acids, etc. and their interactions. These components endow tea wine with unique flavors and tastes. Different types of tea wine may have different characteristics and flavors, which depend on factors such as brewing process, raw materials and storage conditions.

Claims

1. A method for preparing Pu'er tea flavored wine, characterized in that: At least the following steps are included: S1. Pile fermentation: Add a certain amount of water to the Pu'er raw tea residue for wet operation, then add fungi for pile culture fermentation, then add to the steamed and cooled grains, mix well, add yeast, and mix well again; S2, bacterial culture and saccharification: collecting the product obtained in step S1 and cultivating bacteria in a pile, controlling the pile temperature to cultivate bacteria and saccharify to obtain mash; S3, putting the fermented grains into the tank: after the culture and saccharification are completed, the fermented grains are put into the tank for closed fermentation; S4, steaming wine in an upper steamer: steaming the material obtained after the treatment in step S3, collecting the distillate after the steaming to obtain new wine, and then sealing the new wine and storing it for at least one year to obtain Pu'er tea flavored wine.

2. The method for preparing Pu'er tea flavored wine according to claim 1, characterized in that: The fungi include at least one of Aspergillus niger, Aspergillus wusha, Rhizopus niger, Rhizopus oryzae and Rhizopus chinensis.

3. The method for preparing Pu'er tea flavored wine according to claim 2, characterized in that: The fungi are Aspergillus niger and Rhizopus oryzae.

4. The method for preparing Pu'er tea flavored wine according to claim 3, characterized in that: The mass ratio of the Aspergillus niger to the Rhizopus oryzae is 1:(2-5).

5. The method for preparing Pu'er tea flavored wine according to claim 1, characterized in that: The mass of the water is 35-45% of the mass of the Pu'er raw tea residue.

6. The method for preparing Pu'er tea flavored wine according to claim 1, characterized in that: The yeast comprises at least one of Saccharomyces cerevisiae, Hansenula, Pichia pastoris and Pseudomonas aeruginosa.

7. The method for preparing Pu'er tea flavored wine according to claim 6, characterized in that: The yeast accounts for 0.01-0.5% of the mass of the Pu'er raw tea residue.

8. The method for preparing Pu'er tea flavored wine according to claim 1, characterized in that: The culture and saccharification time is 20-24h.

9. The method for preparing Pu'er tea flavored wine according to claim 8, characterized in that: The pile temperature is 25-38°C.

10. The method for preparing Pu'er tea flavored wine according to claim 1, characterized in that: The time for steaming wine in the upper steamer is 60-70 minutes, and the temperature of steaming wine in the upper steamer is 90-110°C.

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