Preparation technology of fermentation type rosa roxburghii tratt fruit and green tea wine and product thereof

Abstract

The invention discloses a preparation process of fermentation type roxburgh rose green tea wine and a product thereof, and relates to the technical field of preparation of fruit tea wine. The preparation process comprises the following steps: preparation of green tea extraction tea soup, component adjustment, yeast activation, fermentation, centrifugation and sterilization. The optimal fermentation process obtained through test investigation is as follows: the fermentation time is 9 days, the initial fermentation sugar degree is 22-degree Brix, the yeast inoculum size is 0.4%, the volume ratio of the roxburgh rose stock solution to the tea soup is 1: 5, the fermentation temperature is 26 DEG C, and the initial fermentation pH is 3.5. According to the fermentation type rosa roxburghii tratt green tea wine prepared under the optimal technological conditions, the alcoholic strength is 12.73% vol, the tea polyphenol content is 702 mg / L, the vitamin C content is 724 mg / L, the wine body is bright in color and rich in rosa roxburghii tratt fruit fragrance and tea fragrance, fragrant substances in the green tea wine are well reserved, ester, alcohol and acid substances of the green tea wine are enriched through introduction of rosa roxburghii tratt, and the taste of the green tea wine is improved. The green tea wine is endowed with more levels of fruit fragrance, grass fragrance, flower fragrance and milk fragrance, the taste is soft, the aftertaste is sweet, and the varieties of roxburgh rose and green tea processed products are enriched.

Description

Technical Field

[0001] The invention relates to the technical field of fruit tea wine preparation, and in particular to a preparation process of fermented roxburghii green tea wine and a product thereof. Background Art

[0002] Tea wine is an alcoholic beverage made mainly from tea, which combines the aroma of tea with the mellowness of wine, showing a unique and charming flavor. Tea has high nutritional value and health benefits because it is rich in nutrients and medicinal ingredients. Fermented tea wine has many potential benefits for human health, including enhancing immunity, protecting nerves, anti-oxidation and antibacterial effects, which has attracted widespread attention from researchers and consumers. [1-3] At present, the tea wines that have been developed include oolong tea wine, green tea wine and black tea wine. [4-5] However, tea wine fermented from tea leaves alone often has the problem of weak flavor and insufficient layers, so tea wine fermented from fruit and tea has become an important development trend. [6] Regarding fruit tea wine, the types and contents of monomer phenols and volatile substances in fermented kiwi black tea wine increased significantly, which improved the flavor and quality of the tea wine. [7] Other fruit tea wines studied include citrus tea wine, sugarcane sweet tea wine, and grape tea wine, but there is no research on fermented roxburghii green tea wine. [8-10] .

[0003] Rosa roxburghii Tratt. is the fruit of the perennial deciduous shrub Rosaceae. As a Chinese specialty fruit resource, its planting area and output in Guizhou rank first.

[11] The roxburghii fruit is rich in nutrients and is a natural precious fruit that can be used as both medicine and food. The functional active ingredients in the fruit, such as VC, flavonoids, polyphenols, polysaccharides, terpenoids and amino acids, give the roxburghii fruit good physiological effects such as anti-oxidation, anti-cancer and anti-atherosclerosis.

[12] Modern research shows that the aroma of roxburghii is special, and the fruit wine brewed with it is clear and stable, with a prominent fruity aroma, which is suitable for fruit wine brewing.

[13] .

[0004] Based on this, the present invention uses roxburgh stock solution and high mountain green tea as main raw materials, ferments roxburgh green tea wine by BV818 active dry yeast, and determines the optimal fermentation process through single factor investigation and orthogonal experiment, in order to develop fermented roxburgh green tea wine with both roxburgh and green tea flavors and nutrition, so as to enrich the types of tea wine products and provide reference and data support for making full use of tea and roxburgh resources. Summary of the invention

[0005] The purpose of the present invention is to provide a preparation process of fermented roxburgh green tea wine and a fermented roxburgh green tea wine product prepared by the preparation process.

[0006] To achieve the above purpose, the technical solution adopted by the present invention is as follows:

[0007] The preparation process of the fermented roxburghii green tea wine of the present invention comprises the following steps:

[0008] S1: preparing green tea extract soup: grinding green tea leaves and mixing with water, subjecting them to microwave treatment, water bath extraction, filtering, and removing the filter residue to prepare green tea extract soup;

[0009] S2 adjusts the ingredients: mix the roxburghii stock solution and the green tea extract in a certain volume ratio, add white sugar to adjust the initial sugar content, add citric acid and baking soda to adjust the initial pH, and add 120 mg / L of potassium metabisulfite to prepare a roxburghii green tea soup preparation;

[0010] S3 Yeast activation: Weigh a certain amount of BV818 active dry yeast and add it to sucrose aqueous solution for constant temperature culture;

[0011] S4 fermentation: put the prepared Rosa roxburghii green tea soup into a sterilized fermentation bottle, add the activated yeast solution, shake well, and ferment in a constant temperature and humidity incubator;

[0012] S5 centrifugation: After stopping the fermentation, centrifuge at 3000r / min for 15min to clarify the wine sample and keep the supernatant;

[0013] S6 sterilization: Use pasteurization, heat in a water bath until the center temperature of the wine reaches 65-70℃, and keep warm for 30 minutes for sterilization.

[0014] The specific process of preparing green tea extraction tea soup in step S1 in the preparation process of the fermented roxburghii green tea wine of the present invention is as follows:

[0015] 1) Take an appropriate amount of green tea leaves, crush them with a tissue crusher, and pass them through a 20-mesh sieve;

[0016] 2) Mix the sieved tea leaves with water at a ratio of 1 g:60 mL, place in a microwave oven, microwave at 720 W for 3 min, and then soak in a 55°C water bath for 25 min;

[0017] 3) Filter with four layers of gauze, remove the filter residue, and obtain green tea infusion.

[0018] In the preparation process of the fermented roxburghii green tea wine of the present invention, the volume ratio of the roxburghii stock solution and the green tea extract in the step S2 is adjusted to 1:2-6, the initial sugar content is 18-26° Brix, and the initial pH is 2.5-4.5.

[0019] Preferably, in the preparation process of the fermented sea buckthorn green tea wine of the present invention, the volume ratio of the sea buckthorn stock solution and the green tea extraction tea soup in step S2 of the adjustment component is 1:4-6, the initial sugar content is 20-24° Brix, and the initial pH is 3-4.5.

[0020] Further preferably, the volume ratio of the roxburghii stock solution to the green tea extract tea soup of the present invention is 1:5, the initial sugar content is 22° Brix, and the initial pH is 3.5.

[0021] The specific process of yeast activation in step S3 of the preparation process of the fermented roxburghii green tea wine of the present invention is as follows: weigh a certain amount of BV818 active dry yeast, add it to 10 mL of 20% sucrose aqueous solution, and culture it in a constant temperature incubator at 30° C. for 40 minutes.

[0022] The specific process of fermentation in step S4 of the preparation process of the fermented roxburghii green tea wine of the present invention is as follows: the roxburghii green tea soup preparation liquid is put into a sterilized fermentation bottle, the activated yeast liquid is added at a yeast inoculation amount of 0.1% to 0.5%, the mixture is shaken well, and the mixture is allowed to ferment in an incubator at a temperature of 22° C. to 30° C. and a relative humidity of 53% for 1 to 11 days.

[0023] Preferably, the specific process of fermentation in step S4 of the preparation process of the fermented sea buckthorn green tea wine of the present invention is: the sea buckthorn green tea soup preparation liquid is loaded into a sterilized fermentation bottle, the activated yeast liquid is added at a yeast inoculation amount of 0.2% to 0.5%, and the mixture is shaken well, and the mixture is allowed to ferment in an incubator at a temperature of 22° C. to 28° C. and a relative humidity of 53% for 6 to 11 days.

[0024] Further preferably, the yeast inoculation amount of the present invention is 0.4%, the temperature is 26° C., and the static fermentation time is 9 days.

[0025] The fermented roxburghii green tea wine of the present invention is prepared by adopting the above-mentioned preparation process.

[0026] Beneficial effects of the present invention:

[0027] The present invention conducts a large number of investigations on factors affecting the quality of the roxburghii green tea wine: (1) By investigating the volume ratio of roxburghii stock solution to tea soup (1:2, 1:3, 1:4, 1:5, 1:6), it is found that when the volume ratio is 1:5, the roxburghii green tea wine has a bright appearance, a harmonious roxburghii fruit aroma and a tea aroma, a relatively sweet taste, a slightly salty and slightly bitter taste, a not obvious astringency, an obvious wine taste, a roxburghii flavor and a tea flavor, a highest sensory score, and a moderate alcohol content. (2) By investigating the fermentation temperature (22°C, 24°C, 26°C, 28°C, 30°C), it is found that when the fermentation temperature is 26°C, the sensory score and the alcohol content reach the highest. At this time, the roxburghii green tea wine has a moderate sweetness and sourness, has the flavors of roxburghii and green tea, has a suitable taste, and has a strong wine taste. (3) By investigating the yeast inoculation amount (0.1%, 0.2%, 0.3%, 0.4%, 0.5%), the results showed that when the yeast inoculation amount was 0.3%, the sensory score was the highest, which was 71.7 points, the alcohol content reached 12.36%vol, and the wine was rich and harmonious. Yeast inoculation amounts of 0.2%, 0.3%, and 0.4% were selected for orthogonal experiments. (4) By investigating the initial pH value (2.5, 3, 3.5, 4, 4.5), the results showed that when the initial pH value was 3.5, the sensory score reached the highest point of 72.47 points, the wine was rich and the flavor was harmonious. (5) By investigating the initial sugar content (18°Brix, 20°Brix, 22°Brix, 24°Brix, 26°Brix), it was found that when the initial sugar content was 22°Brix, the alcohol content reached the highest, the sensory score was 73.53 points, the taste was harmonious, and the wine was rich. The initial sugar content of 22°Brix, 24°Brix, and 26°Brix were selected for orthogonal experiments. (6) By investigating the fermentation time (1-11d), it was found that when the fermentation time was 8d, the alcohol content tended to be stable, the sensory score reached the highest, the sea buckthorn green tea wine had a strong wine flavor and a mellow taste, and the fermentation time of 7d, 8d, and 9d was selected for orthogonal experiments. (7) Through single-factor investigation of the fermentation of sea buckthorn green tea wine and orthogonal experiment for process optimization, the optimal process combination was obtained as follows: fermentation time was 9 days, initial sugar content was 22°Brix, yeast inoculation amount was 0.4%, volume ratio of sea buckthorn concentrate to tea soup was 1:5, fermentation temperature was 26°C, and initial pH value of fermentation was 3.5.

[0028] The sensory score of the roxburghii green tea wine brewed under the optimal process conditions provided by the present invention is 84.2 points, the alcohol content is 12.73% vol, the tea polyphenol content is 702 mg / L, the vitamin C content is 724 mg / 100 mL, the wine body is bright in color, rich in roxburghii fruit and tea aroma, soft in taste, and sweet in aftertaste. At the same time, GC-MS aroma substance analysis shows that the roxburghii green tea wine retains the types of aroma substances in the green tea wine very well, and the introduction of roxburghii enriches the esters, alcohols and acids of the green tea wine, giving the green tea wine more levels of fruity, grassy, ​​floral and milky aromas, enriching the types of tea wine products, especially the types of roxburghii and green tea processed products, providing reference and data support for making full use of tea and roxburghii resources. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 Rosa roxburghii green tea wine fermentation process flow chart

[0030] Figure 2 Effect of fermentation temperature on the quality of roxburghii green tea wine

[0031] Figure 3 Effect of yeast inoculation amount on the quality of roxburghii green tea wine

[0032] Figure 4 Effect of initial pH value on the quality of roxburghii green tea wine

[0033] Figure 5 Effect of initial sugar content on the quality of roxburghii green tea wine

[0034] Figure 6 Effect of fermentation time on the quality of roxburghii green tea wine

[0035] Figure 7 Rosa roxburghii green tea wine produced under optimal process conditions DETAILED DESCRIPTION

[0036] The technical solution of the present invention is described in detail below in conjunction with specific embodiments. The following embodiments are only used for explanation and illustration, and do not constitute a limitation on the technical solution of the present invention.

[0037] Example 1

[0038] A preparation process of fermented roxburghii green tea wine, the specific steps are as follows:

[0039] S1: preparing green tea extract soup: 1) taking an appropriate amount of green tea leaves, crushing them with a tissue crusher, and passing them through a 20-mesh sieve; 2) mixing the sieved tea leaves with water at a ratio of 1 g:60 mL, placing them in a microwave oven, treating them at a microwave power of 720 W for 3 min, and then extracting them in a 55° C. water bath for 25 min; 3) filtering them with four layers of gauze, removing the filter residue, and obtaining green tea extract soup;

[0040] S2: adjusting ingredients: mixing the roxburghii stock solution and the green tea extract in a volume ratio of 1:5, adding white sugar to adjust the initial sugar content to 22° Brix, adding citric acid and baking soda to adjust the initial pH to 3.5, and adding 120 mg / L of potassium metabisulfite to prepare a roxburghii green tea soup preparation;

[0041] S3 Yeast activation: Weigh a certain amount of BV818 active dry yeast, add it to 10mL 20% sucrose aqueous solution, and culture it in a constant temperature incubator at 30℃ for 40min;

[0042] S4 fermentation: put the prepared Rosa roxburghii green tea soup into a sterilized fermentation bottle, add the activated yeast solution at a yeast inoculation rate of 0.4%, shake well, and ferment in an incubator at a temperature of 26°C and a relative humidity of 53% for 9 days;

[0043] S5 centrifugation: After stopping the fermentation, centrifuge at 3000r / min for 15min to clarify the wine sample and keep the supernatant;

[0044] S6 sterilization: Use pasteurization, heat in a water bath until the center temperature of the wine reaches 65-70℃, and keep warm for 30 minutes for sterilization.

[0045] A roxburghii green tea wine product prepared according to the preparation process.

[0046] Example 2

[0047] A preparation process of fermented roxburghii green tea wine, the specific steps are as follows:

[0048] S1: preparing green tea extract soup: 1) taking an appropriate amount of green tea leaves, crushing them with a tissue crusher, and passing them through a 20-mesh sieve; 2) mixing the sieved tea leaves with water at a ratio of 1 g:60 mL, placing them in a microwave oven, treating them at a microwave power of 720 W for 3 min, and then extracting them in a 55° C. water bath for 25 min; 3) filtering them with four layers of gauze, removing the filter residue, and obtaining green tea extract soup;

[0049] S2: adjusting ingredients: mixing the roxburghii stock solution and the green tea extract in a volume ratio of 1:4, adding white sugar to adjust the initial sugar content to 18° Brix, adding citric acid and baking soda to adjust the initial pH to 3, and adding 120 mg / L of potassium metabisulfite to prepare a roxburghii green tea soup preparation;

[0050] S3 Yeast activation: Weigh a certain amount of BV818 active dry yeast, add it to 10 mL of 20% sucrose aqueous solution, and culture it in a constant temperature incubator at 30°C for 40 minutes;

[0051] S4 fermentation: put the prepared Rosa roxburghii green tea soup into a sterilized fermentation bottle, add the activated yeast solution at a yeast inoculation rate of 0.1%, shake well, and ferment in an incubator at a temperature of 28° C. and a relative humidity of 50% for 11 days;

[0052] S5 centrifugation: After stopping the fermentation, centrifuge at 3000r / min for 15min to clarify the wine sample and keep the supernatant;

[0053] S6 sterilization: Use pasteurization, heat in a water bath until the center temperature of the wine reaches 65-70℃, and keep warm for 30 minutes for sterilization.

[0054] A roxburghii green tea wine product prepared according to the preparation process.

[0055] Example 3

[0056] A preparation process of fermented roxburghii green tea wine, the specific steps are as follows:

[0057] S1: preparing green tea extract soup: 1) taking an appropriate amount of green tea leaves, crushing them with a tissue crusher, and passing them through a 20-mesh sieve; 2) mixing the sieved tea leaves with water at a ratio of 1 g:60 mL, placing them in a microwave oven, treating them at a microwave power of 720 W for 3 min, and then extracting them in a 55° C. water bath for 25 min; 3) filtering them with four layers of gauze, removing the filter residue, and obtaining green tea extract soup;

[0058] S2: adjusting ingredients: mixing the roxburghii stock solution and the green tea extract in a volume ratio of 1:6, adding white sugar to adjust the initial sugar content to 26° Brix, adding citric acid and baking soda to adjust the initial pH to 4.5, and adding 120 mg / L of potassium metabisulfite to prepare a roxburghii green tea soup preparation;

[0059] S3 Yeast activation: Weigh a certain amount of BV818 active dry yeast, add it to 10 mL of 20% sucrose aqueous solution, and culture it in a constant temperature incubator at 30°C for 40 minutes;

[0060] S4 fermentation: put the prepared Rosa roxburghii green tea soup into a sterilized fermentation bottle, add the activated yeast solution at a yeast inoculation rate of 0.5%, shake well, and ferment in an incubator at a temperature of 22° C. and a relative humidity of 55% for 6 days;

[0061] S5 centrifugation: After stopping the fermentation, centrifuge at 3000r / min for 15min to clarify the wine sample and keep the supernatant;

[0062] S6 sterilization: Use pasteurization, heat in a water bath until the center temperature of the wine reaches 65-70℃, and keep warm for 30 minutes for sterilization.

[0063] A roxburghii green tea wine product prepared according to the preparation process.

[0064] In order to further verify the reliability of the present invention and screen out the best solution, the inventor conducted a series of experiments, as follows:

[0065] 1. Materials and methods

[0066] 1.1 Materials and reagents

[0067] Seabuckthorn concentrate: Guizhou Qianweipiao Agricultural Development Co., Ltd.; Zhongmin Fengzhou high mountain tea: Fujian Anxi Baiyu Biotechnology Co., Ltd.; Angel wine active dry yeast BV818: Angel Yeast Co., Ltd.; White sugar (food grade): Gengma Nanhua Sugar Co., Ltd.; Baking soda (food grade): Henan Boming Food Co., Ltd.; Citric acid (food grade): Shandong Yingxuan Co., Ltd.; Phenolphthalein, potassium metabisulfite, sodium hydroxide, sodium chloride (all analytical grade): Beijing Wokai Biotechnology Co., Ltd.; Potassium dihydrogen phosphate, oxalic acid, ascorbic acid standards, sodium bicarbonate (all analytical grade): Sinopharm Chemical Reagent Co., Ltd.; Anhydrous ethanol (analytical grade): Shanghai Hushi Laboratory Equipment Co., Ltd.; 2,6-Dichloroindophenol (analytical grade): Shanghai McLean Biochemical Technology Co., Ltd.; Metaphosphoric acid (analytical grade): Tianjin Komiou Chemical Reagent Co., Ltd.

[0068] 1.2 Instruments and Equipment

[0069] Agilent 7000D gas chromatograph-mass spectrometer: Agilent Technologies Co., Ltd.; ST3100 pH meter: Ohaus Instrument (Changzhou) Co., Ltd.; 25mL density bottle with thermometer: Chongming Construction Glass Instrument Co., Ltd.; FA2004W electronic balance: Shanghai Jinghai Instrument Co., Ltd.; LY05-100 constant temperature and humidity incubator: Shanghai Longyue Instrument Equipment Co., Ltd.; BGG-9410A blast drying oven, TD32 handheld sugar meter: Shanghai Yiheng Scientific Instrument Co., Ltd.; CV-5200PC UV spectrophotometer: Shanghai Yuanxi Instrument Co., Ltd.; M3-L205C microwave oven: Guangdong Midea Kitchen Appliance Manufacturing Co., Ltd.; DK-S24 electric constant temperature water bath: Tianjin Test Instrument Co., Ltd.; TDZ5-WS centrifuge: Xiangyi Centrifuge Instrument Co., Ltd.

[0070] 1.3 Methods

[0071] 1.3.1 Rosa roxburghii green tea wine fermentation process and key points of operation

[0072] Process flow, see Figure 1 .

[0073] Operation points:

[0074] (1) Preparation of green tea extract: green tea leaves were crushed with a tissue crusher, passed through a 20-mesh sieve, mixed with water at a ratio of 1:60 and placed in a microwave oven at a microwave power of 720 W for 3 min, extracted in a 55°C water bath for 25 min, and the green tea residue and other sediments were filtered out with four layers of gauze, and the tea extract was retained for later use;

[0075] (2) Adjusting ingredients: preparing the roxburghii green tea soup according to a certain volume ratio of roxburghii stock solution to green tea soup, adding white sugar to adjust the initial sugar content of the fermentation liquid, adding citric acid and baking soda to adjust the pH, and adding 120 mg / L of potassium metabisulfite;

[0076] (3) Yeast activation: Weigh a certain amount of BV818 active dry yeast, add it to 10 mL of 20% sucrose aqueous solution, and culture it in a constant temperature incubator at 30°C for 40 min;

[0077] (4) Fermentation: Pour the prepared Rosa roxburghii green tea soup into a sterilized fermentation bottle, add the activated yeast solution and shake well, and place in a constant temperature and humidity incubator for fermentation;

[0078] (5) Centrifugation: After stopping the fermentation, the wine sample is clarified by centrifugation (3000 r / min, 15 min), and the supernatant is retained;

[0079] (6) Sterilization: Use pasteurization, heat in a water bath until the center temperature of the wine reaches 65-70°C and keep it warm for 30 minutes for sterilization.

[0080] 1.3.2 Single factor experiment on optimization of fermentation process of roxburghii green tea wine

[0081] The single-factor rotation method was adopted, and the basic fermentation conditions were: the volume ratio of sea buckthorn stock solution to tea soup was 1:4, the yeast inoculation amount was 0.3%, the fermentation temperature was 26°C, the initial sugar content was 22°Brix, the fermentation time was 8d, and the initial pH value was 3.5. Based on the basic conditions, the effects of the volume ratio of sea buckthorn concentrate to tea soup (1:2, 1:3, 1:4, 1:5, 1:6), yeast inoculation amount (0.1%, 0.2%, 0.3%, 0.4%, 0.5%), fermentation temperature (22℃, 24℃, 26℃, 28℃, 30℃), initial sugar content (18°Brix, 20°Brix, 22°Brix, 24°Brix, 26°Brix), fermentation time (1d, 2d, 3d, 4d, 5d, 6d, 7d, 8d, 9d, 10d, 11d) and initial pH value (2.5, 3, 3.5, 4, 4.5) on the sensory score and alcohol content of sea buckthorn green tea wine were investigated in turn.

[0082] 1.3.3 Orthogonal experimental design for optimization of the fermentation process of roxburghii green tea wine

[0083] According to the results of the single factor experiment, L9(3 4) Orthogonal table 1 An orthogonal experimental scheme was designed to study the effects of the more significant factors A (fermentation time), B (initial sugar content), C (yeast inoculation amount), and D (volume ratio of sea buckthorn stock solution to tea soup) in the single factor experiment on the fermentation of tea wine. At the same time, the optimal process conditions were determined based on the alcohol content and sensory scores, as shown in Table 1.

[0084] Table 1 Orthogonal test factors and levels for optimization of the fermentation process of roxburghii green tea wine

[0085]

[0086] 1.3.4 Analysis and testing

[0087] Titration acid: Refer to GB / T 15038-2006 "General analysis method for wine and fruit wine" and use the acid-base titration method for determination

[14] ; Alcohol content: Refer to GB 5009.225-2023 "Ethanol concentration in wine" density bottle method for alcohol content determination

[15] ; Tea polyphenols: refer to Appendix A of GB / T 21733-2008 "Tea Beverages"

[16] ; Determination of vitamin C: refer to the 2,6-dichloroindophenol titration method in GB5009.86-2016 "Determination of ascorbic acid in food"

[17] ; Sugar content: measured by handheld sugar meter

[18] .

[0088] Sensory evaluation of roxburgh green tea wine: The sensory evaluation methods of tea wine and fruit wine were combined to formulate the sensory evaluation standards of roxburgh green tea wine, which mainly include appearance, aroma, taste, and typicality. Referring to the sensory evaluation method in the "General Analysis Method for Wine and Fruit Wine", a sensory evaluation team composed of 15 food professionals scored the appearance, aroma, taste, and typicality of roxburgh green tea wine, with a full score of 100 points, and the average score of the 15 people was taken as the final result of the sensory evaluation.

[14] The sensory scoring standard of roxburghii green tea wine is shown in Table 2.

[0089] Table 2 Sensory scoring standards for roxburghii green tea wine

[0090]

[0091]

[0092] GC-MS analysis of aroma components of roxburghii green tea wine, single roxburghii wine, and single green tea wine:

[0093] (1) HS-SPME extraction conditions: Add 7 mL of sample to the headspace bottle, add 1.5 g of sodium chloride to accelerate the salting-out rate, add 10 μL of 2-octanol (56 mg / L) as the internal standard, and tighten the bottle stopper by shaking. Under constant temperature conditions of 45°C, shake for 5 min, insert the 120 μm DVB / CWR / PDMS extraction head into the sample headspace bottle, extract the headspace for 40 min, analyze at 250°C for 5 min, and then perform GC-MS separation and identification. Before sampling, the extraction head was aged at 250°C for 5 min in the Fiber Conditioning Station.

[0094] (2) Chromatographic conditions: DB-FFAP capillary column (60m×0.25mm×0.25μm), carrier gas: high-purity helium, constant flow rate: 1.2mL / min, injection port temperature: 250℃, splitless injection, solvent delay: 3.0min. Program temperature: 40℃ to 50℃ at 10℃ / min, hold for 2min, then to 80℃ at 3℃ / min, hold for 2min, and finally to 245℃ at 5℃ / min, hold for 3min.

[0095] (3) Mass spectrometry conditions: electron impact ion source (EI), ion source temperature 230°C, quadrupole temperature 150°C, mass spectrometry interface temperature 250°C, electron energy 70 eV, full scan scanning mode, range 10 amu-550 amu.

[0096] (4) Qualitative and quantitative analysis: The ion spectra obtained by analysis were searched through the Wiley spectral library and the National Institute of Standards and Technology (NIST) 2.0 spectral library, and only substances with a matching degree of more than 80 were retained, and quantification was performed using the internal standard method.

[0097] 1.3.5 Data processing

[0098] GraphP Prism software was used to draw graphs, and SPSS software was used for one-way analysis of variance. P < 0.05 indicated that the results were statistically significant. All experiments were carried out in parallel three times, and the test results were expressed as "mean ± standard deviation".

[0099] 2. Results and Analysis

[0100] 2.1 Results of single factor experiment on optimization of fermentation process of roxburghii green tea wine

[0101] 2.1.1 Effect of the volume ratio of roxburghii stock solution to tea soup on the quality of roxburghii green tea wine

[0102] The results showed that as the volume ratio of roxburghii stock solution to tea soup varied from 1:2 to 1:6, the sensory score increased first and then decreased, and the alcohol content showed a gradually decreasing trend. The volume ratio of roxburghii stock solution to tea soup had a significant effect on the sensory score and alcohol content (P < 0.05). When the volume ratio of roxburghii stock solution to tea soup was 1:2 and 1:3, the sensory score was low, the wine was dark yellow, the roxburghii flavor was too strong, the tea aroma was masked, and the sour taste was obvious; when the volume ratio of roxburghii stock solution to tea soup was 1:4, the wine was bright yellow, clear and transparent, with the main roxburghii aroma and weak tea aroma; when the volume ratio of roxburghii stock solution to tea soup was 1:5 and 1:6, the wine was bright yellow, clear and transparent, with the roxburghii aroma and tea aroma coordinated. When the volume ratio of roxburghii stock solution to tea soup was 1:6, the wine was slightly sweet and the alcohol content was lower than others (10.42% vol). After comprehensive analysis, the volume ratios of 1:4, 1:5, and 1:6 of the roxburghii stock solution to tea soup were selected for orthogonal test. The results are shown in Table 3.

[0103] Table 3 Effect of the volume ratio of roxburghii stock solution to tea soup on the quality of roxburghii green tea wine

[0104]

[0105] Note: The data in the table represent mean ± standard deviation (N = 3), and lowercase letters and uppercase letters represent the significance of sensory scores and alcohol content, respectively (P < 0.05, Tukey method).

[0106] 2.1.2 Effect of fermentation temperature on the quality of roxburghii green tea wine

[0107] The results showed that with the increase of fermentation temperature in the range of 22-30℃, the sensory score and alcohol content of the roxburghii green tea wine showed a trend of first rising and then falling, and the fermentation temperature had a significant effect on the sensory score and alcohol content (P<0.05). The fermentation temperature will affect the growth and metabolism of yeast. High or low temperature is not conducive to the production and conversion of alcohol, thus affecting the sensory quality of the fruit wine. The sensory score and alcohol content reached the highest when the fermentation temperature was 26℃, which were 80.41 points and 12.28%vol, respectively. At this time, the roxburghii green tea wine had moderate sweetness and sourness, with the flavor of roxburghii and green tea, suitable taste, and strong wine flavor. The sensory scores and alcohol content of the fermentation temperatures of 22℃, 24℃, and 28℃ were all lower than 26℃. When the fermentation temperature was 30℃, the sensory score and alcohol content were the lowest, and the roxburghii green tea wine had a dark color and a heavy bitter taste. The fermentation temperature was high, yeast was intolerant, and metabolism was inhibited, and the alcohol content decreased. Comprehensive analysis, the initial fermentation temperature was selected as 26℃. The results of the effect of fermentation temperature on the quality of roxburghii green tea wine are shown in Figure 2 The bar graph data in the figure represent the mean ± standard deviation (N = 3), and the lowercase letters and uppercase letters represent the significance of sensory scores and alcohol content, respectively (P < 0.05, Tukey method), the same below.

[0108] 2.1.3 Effect of yeast inoculation amount on the quality of roxburghii green tea wine

[0109] The results showed that as the yeast inoculation increased from 0.1% to 0.5%, the sensory score of the roxburghii green tea wine showed a trend of first increasing and then decreasing, and the alcohol content showed a trend of first increasing and then stabilizing, and the yeast inoculation had a significant effect on the sensory score and alcohol content (P < 0.05). During the fermentation of fruit wine, the yeast inoculation will affect the yeast growth rate, fermentation rate and respiratory capacity. When the yeast inoculation was 0.1%, the sensory score and alcohol content were low, which may be due to the small yeast inoculation, resulting in insufficient fermentation, slow fermentation rate and low alcohol conversion rate; when the yeast inoculation was 0.3%, the sensory score was the highest, which was 78.87 points, the alcohol content reached 12.36% vol, and the wine body was coordinated and rich; when the yeast inoculation increased to 0.5%, the alcohol content increased accordingly, but the yeast respiratory capacity accelerated, and the growth and reproduction were too fast, which would lead to the oxidation of some esters, alcohols and other substances, thereby affecting the taste and flavor of the roxburghii fruit wine, and the sensory score decreased. After comprehensive analysis, yeast inoculations of 0.2%, 0.3% and 0.4% were selected for orthogonal experiments. Results Figure 3 .

[0110] 2.1.4 Effect of initial pH value on the quality of roxburghii green tea wine

[0111] The results showed that as the initial pH value increased from 2.5 to 4.5, the sensory score and alcohol content of the roxburghii green tea wine showed a trend of first increasing and then decreasing, and the yeast inoculation amount had a significant effect on the sensory score and alcohol content (P < 0.05). The pH of the fermentation liquid has a certain effect on the growth and metabolism of yeast. When the pH is too low, the yeast's tolerance to CO2 decreases, which inhibits yeast reproduction; when the pH is too high, the yeast metabolism and alcohol production capacity weaken, which easily leads to bacterial infection in the fermentation liquid. When the initial pH value of the fermentation liquid is 2.5 and 3.0, the alcohol content is 0.02% vol and 9.69% vol, the wine is not thick enough, and the sour taste is strong; when the initial pH value of the fermentation liquid is 4.5, the alcohol production decreases, the off-flavor is strong, and the color is dark. When the pH of the fermentation liquid is 3.5 and 4.0, the yeast grows and reproduces well. When the initial pH value is 4.0, the alcohol content reaches the highest of 12.28% vol, and when the initial pH value is 3.5, the sensory score reaches the highest of 79.71, the wine is rich and the flavor is coordinated. After comprehensive analysis, the initial pH value was selected to be 3.5. Figure 4 .

[0112] 2.1.5 Effect of initial sugar content on the quality of roxburghii green tea wine

[0113] The results showed that as the initial sugar content increased from 18 to 26°Brix, the sensory score and alcohol content both showed a trend of first increasing and then decreasing, and the initial sugar content had a significant effect on the sensory score and alcohol content (P < 0.05). Sugar is the main fermentation substance for yeast fermentation and the main source of alcohol. If the sugar content is too low, the fermentation source will be insufficient, which will lead to a decrease in alcohol production, while too high a sugar content will inhibit the growth of yeast and lead to a decrease in alcohol content. When the initial sugar content was 18°Brix, the sensory score was low (71.72 points), the sea buckthorn green tea wine tasted sour, the tea flavor was light, and the taste was thin; when the initial sugar content was 22°Brix, the alcohol content reached a maximum of 12.26%vol, the sensory score was 80.89 points, the taste was coordinated, and the wine was full-bodied; when the initial sugar content was 24°Brix, the alcohol content began to decrease, and the sensory score was the highest (81.95 points), the taste was coordinated, the wine was full-bodied, the sourness and sweetness were moderate, and the taste was better; when the initial sugar content continued to increase to 26°Brix, the alcohol content and sensory score both decreased. The excessively high sugar content made the sea buckthorn green tea wine taste sweet and greasy, and not coordinated enough. After comprehensive analysis, orthogonal experiments were conducted with initial sugar contents of 22°Brix, 24°Brix, and 26°Brix. Results are shown in Figure 5 .

[0114] 2.1.6 Effect of fermentation time on the quality of roxburghii green tea wine

[0115] The results showed that as the fermentation time increased from 1d to 11d, the alcohol content first increased rapidly and then stabilized, and the sensory score first increased and then decreased. The fermentation time had a significant effect on both the sensory score and the alcohol content (P < 0.05). The change in alcohol content corresponds to the growth curve of the yeast. In the early stage of fermentation, the sugar source is sufficient, the yeast grows rapidly, and the alcohol content increases rapidly. In the late stage of fermentation, the yeast enters the decline period, and the alcohol content increases slowly or almost does not increase. When the fermentation time is 1d to 5d, the fermentation time is too short, the fermentation is incomplete, and the flavor is poor; when the fermentation time is 8d, the alcohol content stabilizes (12.26% vol), and the sensory score reaches the highest 80.63 points. At this time, the sea buckthorn green tea wine has a strong aroma and a mellow taste; when the fermentation time continues to increase, the alcohol content rises slowly, and the sensory score begins to decrease. At this time, the astringency becomes heavier, which may be due to the decomposition of ester substances, the increase of sourness and other miscellaneous substances, making the taste inconsistent and the flavor affected. Results are shown in Figure 6 .

[0116] 2.2 Orthogonal test results for optimization of the fermentation process of roxburghii green tea wine

[0117] 2.2.1 Orthogonal experimental design and results

[0118] The orthogonal experimental design and results of the optimization of the fermentation process of sea buckthorn green tea wine are shown in Table 4.

[0119] Table 4 Orthogonal test results for optimization of fermentation process of roxburghii green tea wine

[0120]

[0121]

[0122] Note: The data in the table represent mean ± standard deviation (N = 3).

[0123] As can be seen from Table 4, in the orthogonal test results, when alcohol content was used as the evaluation index, the order of influence of each factor on the alcohol content of sea buckthorn green tea wine was yeast inoculation amount (C)> fermentation time (A)> initial sugar content (B)> volume ratio of sea buckthorn stock solution to tea soup (D), and the optimal fermentation process was A3B1C3D3; when sensory score was used as the evaluation index, the order of influence of each factor on the sensory score of sea buckthorn green tea wine was yeast inoculation amount (C)> fermentation time (A)> volume ratio of sea buckthorn stock solution to tea soup (D)> initial sugar content (B), and the optimal fermentation process was A3B1C3D2. Through the variance analysis of Tables 5 and 6, it can be seen that the factors of fermentation time (A) and yeast inoculation amount (C) have extremely significant effects on the alcohol content of sea buckthorn green tea wine (P < 0.01), and the factors of initial sugar content (B) and the volume ratio of sea buckthorn stock solution to tea soup (D) have significant effects on the alcohol content of sea buckthorn green tea wine (P < 0.05); the factors of fermentation time (A) and yeast inoculation amount (C) have significant effects on the sensory score of sea buckthorn green tea wine (P < 0.05), and the factors of initial sugar content (B) and the volume ratio of sea buckthorn stock solution to tea soup (D) have no significant effects on the sensory score of sea buckthorn green tea wine.

[0124] Table 5 Analysis of variance of orthogonal test factors of roxburghii green tea wine fermentation (alcohol content)

[0125]

[0126]

[0127] Note: In the table, * indicates significant effect (P < 0.05); ** indicates extremely significant effect (P < 0.01), the same below.

[0128] Table 6 Analysis of variance table of orthogonal test factors of roxburghii green tea wine fermentation (sensory score)

[0129]

[0130] After comprehensive consideration, A3B1C3D3 and A3B1C3D2 were selected as the optimal process combination alternatives. A3B1C3D3 had a fermentation time of 9 days, a fermentation sugar content of 22°Brix, a yeast inoculation amount of 0.4%, and a volume ratio of 1:6 for the roxburghii stock solution. A3B1C3D2 had a fermentation time of 9 days, a fermentation sugar content of 22°Brix, a yeast inoculation amount of 0.4%, and a volume ratio of 1:5 for the roxburghii stock solution. Further verification experiments are needed for A3B1C3D3.

[0131] 2.2.2 Experimental verification of the best process of orthogonal optimization

[0132] Through the confirmatory test, it was determined that the optimal combination of roxburghii green tea wine was A3B1C3D2, that is, the optimal process conditions of the optimal roxburghii green tea wine were: fermentation time was 9d, initial sugar content was 22°Brix, yeast inoculation amount was 0.4%, the volume ratio of roxburghii stock solution to tea soup was 1:5, fermentation temperature was 26℃, and the initial pH value of fermentation was 3.5. The alcohol content of roxburghii green tea wine under this process was 12.73%vol, and the sensory score was 84.2 points. The roxburghii green tea wine obtained under the optimal fermentation process conditions was bright and shiny, with a harmonious and rich roxburghii fruit aroma and tea aroma, a soft taste, a full body and a sweet aftertaste. At the same time, the physical and chemical indicators of the roxburghii green tea wine were tested, and the titrated acid content was 6.43±0.07g / L, the tea polyphenol content was 702±16.33mg / L, and the vitamin C content was 724±9.33mg / 100mL. Its sensory requirements, alcohol content, vitamin C, and titrated acid all met the standards of roxburghii wine. The results are shown in Table 7.

[0133] Table 7 Rosa roxburghii green tea wine verification test results

[0134]

[0135] 2.2.3 Comparative analysis of volatile aroma components of roxburghii green tea wine, roxburghii wine alone, and green tea wine alone

[0136] The results showed that 47 aroma components were isolated from roxburghii green tea wine, single roxburghii wine, and single green tea wine, of which 34 were detected in roxburghii green tea wine, 36 were detected in single roxburghii wine, and 24 were detected in single green tea wine. Roxburghii enriched the aroma substances of green tea wine. The aroma components mainly included esters, alcohols, acids, and phenols. The relative content of the above four types of substances accounted for more than 95% of the total aroma components. Isoamyl acetate and linalool, the aroma substances characteristic of green tea, were detected in single green tea wine, and ethyl caproate, ethyl octanoate, and octanoic acid, the aroma substances characteristic of roxburghii, were detected in single roxburghii wine. Both green tea wine and roxburghii wine have the aroma characteristics of green tea and roxburghii.

[0137] Esters are important components of the aroma substances of tea wine and fruit wine, and are mainly produced during the fermentation and aging process. A total of 21 esters were detected in the three fermented wines, of which 15 were found in the roxburghii green tea wine, with a relative content of 68.06%; 18 were found in the single roxburghii wine, with a relative content of 71.13%; and 13 were found in the single green tea wine, with a relative content of 73.07%. Esters contributed the most to the aroma components in the wine samples. The esters shared by the three fermented wines were ethyl acetate, ethyl caproate, isopentyl acetate, ethyl caprylate, phenylethyl acetate, ethyl butyrate, ethyl caprylate, ethyl 3-hexenoate, ethyl phenylacetate, ethyl laurate, and ethyl palmitate. Most of the esters had fruity aroma and unique aromatic odor. Compared with green tea wine alone, more ester substances were detected in the roxburghii green tea wine, including leaf acetate, hexyl formate, ethyl 2-methylbutyrate and ethyl benzoate with apple and grass aromas, of which leaf acetate and ethyl benzoate were derived from roxburghii, and ester substances in green tea wine were retained in the roxburghii green tea wine except for ethyl isovalerate and propyl nonanolide, which were lost. Therefore, roxburghii gives tea wine more ester substances.

[0138] Among the alcohol substances, 13 alcohol substances were detected in the three fermented wines, of which 11 were detected in the roxburghii green tea wine, with a relative content of 5.19%; 9 were detected in the single roxburghii wine, with a relative content of 7.97%; and 5 were detected in the single green tea wine, with a relative content of 2.95%. Roxburghii increased the proportion of alcohol substances in the aroma components of green tea wine. The alcohol substances shared by the three fermented wines are linalool, 2-nonyl alcohol, alpha-terpineol, and trans-nerolidol, which mainly give the wine a floral aroma. Compared with single green tea wine, more alcohol substances were detected in the roxburghii green tea wine, including trans-3-hexen-1-ol, 4-methyl-1-hexanol, 6-methyl-5-heptene-2-ol, 3-methylthiopropanol, 1-nonanol and n-decanol with rose citrus, sweet and floral aromas, and trans-3-hexen-1-ol, 3-methylthiopropanol and 1-nonanol were derived from roxburghii, while all alcohol substances in tea wine were retained in roxburghii green tea wine. Therefore, roxburghii gives tea wine more alcohol substances.

[0139] Acidic substances have a balancing and coordinating effect on the flavor of the wine. A total of 7 acidic substances were detected in the three fermented wines, of which 6 were found in the roxburghii green tea wine, with a relative content of 15.12%; 4 were found in the single roxburghii wine, with a relative content of 12.78%; and 4 were found in the single green tea wine, with a relative content of 14.21%. The proportion of acidic substances in roxburghii green tea wine did not change much compared with that in green tea wine. The alcoholic substances common to the three fermented wines are caprylic acid, decanoic acid and isobutyric acid, which mainly give the wine a milky and fruity aroma. Compared with single green tea wine, more acidic substances were detected in roxburghii green tea wine, including 2-methylbutyric acid and nonanoic acid with cheese aroma. At the same time, all acidic substances in tea wine were retained in roxburghii green tea wine.

[0140] Among other substances, phenols occupy a major position, and a small amount of aldehydes and ketones give the wine a mixed aroma of grassy, ​​coffee and guaiacol. All phenols and ketones in tea wine are retained in sea buckthorn green tea wine, while these substances in sea buckthorn wine are not well retained except for 2,4-di-tert-butylphenol.

[0141] Comprehensive analysis shows that except for the loss of ethyl isovalerate and γ-nonanolide in ester substances, the roxburghii green tea wine retains most of the aroma substances in green tea wine. At the same time, the introduction of roxburghii enriches the esters, alcohols and acids in green tea wine, giving green tea wine more layers of fruity, grassy, ​​floral and milky aromas. The results are shown in Table 8.

[0142] Table 8 Analysis of aroma components of roxburghii green tea wine, single roxburghii wine, and single tea wine

[0143]

[0144]

[0145] Table 8 Analysis of aroma components of roxburghii green tea wine, single roxburghii wine, and single tea wine

[0146]

[0147]

[0148] Note: “a”, aroma characteristics refer to http: / / www.Thegoodscentscompany.com / and http: / / www.chemicalbook.com / ; the data in the table represent mean ± standard deviation (N=3), “-” means the substance was not detected.

[0149] 3. Conclusion

[0150] In this study, sea buckthorn was introduced into tea wine composite fermentation sea buckthorn green tea wine. The optimal fermentation process was obtained through single factor investigation and orthogonal test: fermentation time was 9d, fermentation sugar content was 22°Brix, yeast inoculation amount was 0.4%, volume ratio of sea buckthorn stock solution to tea soup was 1:5, fermentation temperature was 26℃, and initial pH value of fermentation was 3.5. The sensory score of sea buckthorn green tea wine brewed under this process was 84.2 points, alcohol content was 12.73%vol, tea polyphenol content was 702mg / L, vitamin C content was 724mg / 100mL, the wine had bright color, rich sea buckthorn fruit aroma and tea aroma, soft taste, and sweet aftertaste. The product prepared according to the above optimal preparation process is shown in Figure 7At the same time, GC-MS aroma substance analysis showed that the roxburghii green tea wine retained the types of aroma substances in green tea wine very well. The introduction of roxburghii enriched the esters, alcohols and acids in the green tea wine, giving the green tea wine more layers of fruity, grassy, ​​floral and milky aromas.

[0151] Although the present invention has been described in detail above by means of general description, specific implementation methods and experiments, it is obvious to those skilled in the art that some modifications or improvements can be made to the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

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Claims

1. A preparation process of fermented roxburghii green tea wine, characterized in that: The steps include: S1: preparing green tea extract soup: grinding green tea leaves and mixing with water, subjecting them to microwave treatment, water bath extraction, filtering, and removing the filter residue to prepare green tea extract soup; S2 adjusts the ingredients: mix the roxburghii stock solution and the green tea extract in a certain volume ratio, add white sugar to adjust the initial sugar content, add citric acid and baking soda to adjust the initial pH, and add 120 mg / L of potassium metabisulfite to prepare a roxburghii green tea soup preparation; S3 Yeast activation: Weigh a certain amount of BV818 active dry yeast and add it to sucrose aqueous solution for constant temperature culture; S4 fermentation: put the prepared Rosa roxburghii green tea soup into a sterilized fermentation bottle, add the activated yeast solution, shake well, and ferment in a constant temperature and humidity incubator; S5 centrifugation: After stopping the fermentation, centrifuge at 3000r / min for 15min to clarify the wine sample and keep the supernatant; S6 sterilization: Use pasteurization, heat in a water bath until the center temperature of the wine reaches 65-70℃, and keep warm for 30 minutes for sterilization.

2. The preparation process according to claim 1, characterized in that: The specific process of preparing green tea extraction soup in step S1 is as follows: 1) Take an appropriate amount of green tea leaves, crush them with a tissue crusher, and pass them through a 20-mesh sieve; 2) Mix the sieved tea leaves with water at a ratio of 1 g:60 mL, place in a microwave oven, microwave at 720 W for 3 min, and then soak in a 55°C water bath for 25 min; 3) Filter with four layers of gauze, remove the filter residue, and obtain green tea infusion.

3. The preparation process according to claim 1, characterized in that: Step S2 adjusts the volume ratio of the roxburghii stock solution and the green tea extract in the ingredients to 1:2-6, the initial sugar content to 18-26° Brix, and the initial pH to 2.5-4.

5.

4. The preparation process according to claim 3, characterized in that: Step S2 adjusts the volume ratio of the roxburghii stock solution and the green tea extract in the ingredients to 1:4-6, the initial sugar content to 20-24° Brix, and the initial pH to 3-4.

5.

5. The preparation process according to claim 4, characterized in that: The volume ratio of the roxburghii stock solution to the green tea extract is 1:5, the initial sugar content is 22° Brix, and the initial pH is 3.

5.

6. The preparation process according to claim 1, characterized in that: Step S3: The specific process of yeast activation is as follows: a certain amount of BV818 active dry yeast is weighed, added to 10 mL of 20% sucrose aqueous solution, and cultured in a constant temperature incubator at 30° C. for 40 minutes.

7. The preparation process according to claim 1, characterized in that: The specific process of fermentation in step S4 is as follows: the prepared solution of sea buckthorn green tea soup is put into a sterilized fermentation bottle, the activated yeast solution is added at a yeast inoculation rate of 0.1% to 0.5%, the mixture is shaken well, and the mixture is fermented in an incubator at a temperature of 22° C. to 30° C. and a relative humidity of 53% for 1 to 11 days.

8. The preparation process according to claim 7, characterized in that: The specific process of fermentation in step S4 is as follows: the prepared solution of sea buckthorn green tea soup is put into a sterilized fermentation bottle, the activated yeast solution is added at a yeast inoculation rate of 0.2% to 0.5%, the mixture is shaken well, and the mixture is fermented in an incubator at a temperature of 22° C. to 28° C. and a relative humidity of 53% for 6 to 11 days.

9. The preparation process according to claim 8, characterized in that: The yeast inoculation amount is 0.4%, the temperature is 26° C., and the static fermentation time is 9 days.

10. A fermented roxburghii green tea wine, characterized in that: The method is prepared by the preparation process described in any one of claims 1 to 9.

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