CN113180127A - Vine tea and preparation method thereof - Google Patents
Vine tea and preparation method thereof Download PDFInfo
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- CN113180127A CN113180127A CN202110427793.5A CN202110427793A CN113180127A CN 113180127 A CN113180127 A CN 113180127A CN 202110427793 A CN202110427793 A CN 202110427793A CN 113180127 A CN113180127 A CN 113180127A
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Abstract
The invention discloses a preparation method of vine tea, which comprises the following steps: the method comprises the steps of pretreatment, picking, enzyme deactivation, sterilization, fermentation and drying, wherein in the fermentation step, a selenium-rich fermentation mixed solution which takes sodium selenite, glucose, corn flour, water, ammonium sulfate, cyclocarya paliurus extract, plantain extract and zymocyte as raw materials is adopted. The vine tea has good free radical scavenging capacity and blood fat reducing capacity.
Description
Technical Field
The invention belongs to the technical field of tea processing, and particularly relates to vine tea and a preparation method thereof.
Background
The ampelopsis grossedentata is processed from tender stem buds or leaves of Ampelopsis grossedentata of Vitaceae, has multiple effects of inhibiting bacteria, clearing heat and toxic materials, preventing cold, resisting cancer, relieving pain, resisting oxidation, protecting liver, reducing blood sugar and blood fat and the like, and is popular in Hubei, Guangdong, Sichuan, Guizhou and other places. The ampelopsis grossedentata not only contains organic active ingredients such as protein, flavone, polysaccharide, organic acid, steroid and the like, but also contains trace elements required by human bodies such as manganese, copper, iron, zinc, selenium and the like, wherein the selenium is an important trace element for human survival and participates in regulation and control of antioxidant defense, cell growth and apoptosis processes, hormone generation, immunity generation and the like, and in addition, the selenium is also an important vital element for cancer prevention, aging resistance and virus resistance. Therefore, how to improve the selenium content in the ampelopsis grossedentata and the health care and nutritional value of the ampelopsis grossedentata has important significance.
Disclosure of Invention
Aiming at the defects in the prior art, the invention provides vine tea and a preparation method thereof.
In order to solve the technical problems, the invention adopts the technical scheme that:
a preparation method of vine tea comprises the following steps:
s1, pretreatment: controlling the growth temperature of the Ampelopsis grossedentata tea trees to be 20-30 ℃ 25-30 days before the Ampelopsis grossedentata tea leaves are picked, and spraying the selenium-rich nutrient solution on the Ampelopsis grossedentata tea leaves at 4-6 pm every other day, wherein the spraying amount is 60-120L per mu;
s2, picking: picking fresh Ampelopsis grossedentata leaves sprayed with selenium-rich nutrient solution, cleaning with clear water, spreading in shade, and standing for 20-40 hr to obtain pretreated Ampelopsis grossedentata leaves;
s3, fixation: placing the pretreated vine tea leaves in a water-removing machine with the temperature of 130-;
s4, sterilization: sterilizing the deactivated vine tea leaves for 2-5min by steam at the temperature of 100-;
s5, fermentation: soaking sterilized vine tea leaves in a selenium-rich fermentation mixed solution, wherein the mass ratio of the vine tea leaves to the selenium-rich fermentation mixed solution is 1 (10-25), and fermenting for 2-8 days at a relative humidity of 60-70% and a temperature of 30-55 ℃ to obtain fermented vine tea leaves;
s6, drying: drying the fermented Ampelopsis Grossdentata leaves at 35-55 deg.C under relative humidity of 60-80%, turning over the leaves every 0.5-1.5 hr until the water content is 3-7 wt% to obtain Ampelopsis Grossdentata tea.
The preparation method of the selenium-rich nutrient solution in the step S1 is as follows: mixing 2-7 parts by weight of sodium selenite, 1-3 parts by weight of ferrous sulfate, 5-18 parts by weight of citric acid, 10-20 parts by weight of monopotassium phosphate, 5-12 parts by weight of selenomethionine, 0.5-2 parts by weight of absorption promoter and 80-120 parts by weight of water, and stirring at 100-300rpm at room temperature for 2-5min to obtain the selenium-rich nutrient solution.
The first core point of the invention is that the pretreatment is carried out before picking, selenium-rich nutrient solution is sprayed on the fresh leaves of the Ampelopsis grossedentata, and inorganic selenium salt is directly sprayed, so that the inorganic selenium salt is not easy to be absorbed and transformed by plants, a large amount of sodium selenite which is not absorbed can pollute soil, water and the surrounding environment, in addition, the sodium selenite has toxicity, and the sodium selenite which is remained on the surfaces of the plants can cause certain damage to the health of people.
Therefore, the idea of the invention is to convert inorganic selenium salt into organic selenium which is easy to be absorbed by plants, and improve the utilization rate of the fresh leaves of the ampelopsis grossedentata to selenium. Specifically, citric acid is added into the selenium-rich nutrient solution, under the acidic condition of the citric acid, the citric acid is complexed with sodium selenite to obtain organic selenium, under the action of an absorption promoting agent octyl glucoside, the surface of the leaf surface is wetted and permeated, ferrous sulfate can increase the permeability of cells, capillary pores of the leaf surface can be opened, the absorption of the selenium-rich nutrient solution is promoted, and the content of organic selenium in the fresh leaves of the Ampelopsis grossedentata is increased.
The absorption promoting agent is octyl glucoside or polyvinyl alcohol; preferably, the absorption enhancer is octyl glucoside.
The preparation method of the selenium-rich fermentation mixed liquid comprises the following steps: uniformly mixing 2-5 parts by weight of sodium selenite, 5-15 parts by weight of glucose, 1-6 parts by weight of corn flour, 400 parts by weight of 100-plus-one water, 2-6 parts by weight of ammonium sulfate, 1-3 parts by weight of potassium dihydrogen phosphate and 5-10 parts by weight of cyclocarya paliurus extract, sterilizing at the temperature of 100-plus-one water and 130 ℃ for 20-50min, cooling to room temperature to obtain a selenium-rich culture medium, adding 5-10 wt% of zymogen of the selenium-rich culture medium, and culturing at the temperature of 25-35 ℃ and the pH value of 4.0-7.0 for 16-24h to obtain the selenium-rich fermentation mixed solution.
Preferably, the preparation method of the selenium-rich fermentation mixed solution comprises the following steps: uniformly mixing 2-5 parts by weight of sodium selenite, 5-15 parts by weight of glucose, 1-6 parts by weight of corn flour, 400 parts by weight of 100-plus-one water, 2-6 parts by weight of ammonium sulfate, 1-3 parts by weight of potassium dihydrogen phosphate, 5-10 parts by weight of cyclocarya paliurus extract and 1-5 parts by weight of plantain extract, sterilizing at the temperature of 100-plus-one 130 ℃ for 20-50min, cooling to room temperature to obtain a selenium-rich culture medium, adding 5-10 wt% of zymophyte of the selenium-rich culture medium, and culturing at the temperature of 25-35 ℃ and the pH of 4.0-7.0 for 16-24h to obtain the selenium-rich fermentation mixed solution.
The zymocyte is eurotium cristatum selenium-rich and/or orange thermoascus thermophilus; preferably, the fermentation bacteria are prepared from eurotium cristatum selenium-rich and Thermoascus aurantiacus according to the mass ratio of (1-5): 1.
The second core point of the invention is that the selenium content and the antioxidant polysaccharide content in the vine tea are further improved in the fermentation process, the sodium selenite solution on the tea leaves cannot be fully contacted with the eurotium cristatum selenium-rich in the pretreatment spraying selenium-rich nutrient solution, and the eurotium cristatum selenium-rich has the advantages of promoting the utilization rate of the vine tea on organic selenium, improving the yield of converting inorganic selenium into organic selenium, greatly improving the bioavailability of the selenium and improving the utilization rate of raw materials.
The plantain herb is used as a traditional Chinese medicine, can be used for health-care food, has the effects of clearing heat, promoting urination, improving eyesight, protecting liver, reducing blood pressure, inhibiting bacteria, reducing serum cholesterol and the like, and is rich in active ingredients such as plant active polysaccharide, plant polyphenol, trace elements and the like. The cyclocarya paliurus contains abundant flavone, polysaccharide, potassium, calcium, magnesium, phosphorus, iron, copper, chromium, zinc, selenium and other trace elements, the flavone and the polysaccharide in the extract have obvious activities of reducing blood sugar, removing free radicals, resisting oxidation and resisting bacteria, the Eurotium cristatum is added in the fermentation process to enrich selenium, inorganic selenium in the cyclocarya paliurus can be converted into organic selenium, meanwhile, the cyclocarya paliurus extract can be promoted to fully enter the vine tea, and the vine tea is soaked together with effective components in the vine tea when the vine tea is brewed, so that the health care effect is improved.
The preparation method of the cyclocarya paliurus extracting solution comprises the following steps: washing cyclocarya paliurus leaves with water, drying in the air, crushing, sieving with a 30-60-mesh sieve to obtain cyclocarya paliurus leaf powder, taking 40-60 parts by weight of cyclocarya paliurus leaf powder, adding 80-150 parts by weight of water and 0.1-1 part by weight of cellulase, adjusting the pH to 4.5-6, stirring at 200-40 ℃ and 500rpm for 3-10min, extracting at 40-50 ℃ and ultrasonic frequency of 20-40kHz for 20-50min, filtering to obtain a primary filtrate and a primary filter residue, adding a citric acid ethanol solution with the mass of 5-15 wt% of the primary filter residue into the primary filter residue, stirring at 45-55 ℃ and 200-400rpm for 3-5h, filtering to obtain a secondary filtrate and a secondary filter residue, merging the primary filtrate and the secondary filtrate, concentrating by rotary evaporation to 1/20-1/10 of the original mass to obtain cyclocarya paliurus extract.
The preparation method of the plantain herb extracting solution comprises the following steps: cleaning herba plantaginis, air drying, pulverizing, and sieving with 20-80 mesh sieve to obtain herba plantaginis powder; adding 50-60 parts by weight of plantain powder into 80-100 parts by weight of water, uniformly mixing, and performing microwave treatment for 2-6min under the conditions of 200-300W; cooling to room temperature, adding 0.2-0.6 weight part of cellulase and 0.1-0.3 weight part of papain, uniformly mixing, adjusting the pH to 5-7, stirring at 50-100rpm at 40-50 ℃ for 20-30min, filtering to obtain a filtrate I and a filter residue I, adding 60-70 wt% of ethanol aqueous solution into the filter residue I, carrying out reflux extraction at 80-90 ℃ for 150-250min, wherein the mass ratio of the filter residue I to the ethanol aqueous solution is 1 (2-7), filtering to obtain a filtrate II and a filter residue II, combining the filtrate I and the filtrate II, and carrying out rotary evaporation and concentration to 1/10-1/5 of the original mass to obtain the plantain herb extract.
The conditions of the rotary evaporation are as follows: absolute pressure of 0.04-0.06MPa, temperature of 45-55 deg.C, and rotation speed of 30-60 rpm.
A vine tea is prepared by the above method.
The invention has the beneficial effects that: the vine tea of the invention contains rich selenium, and has good free radical scavenging ability and blood fat reducing ability. According to the invention, selenium-rich nutrient solution is sprayed on the fresh ampelopsis grossedentata leaves before picking, so that the selenium absorption rate of the fresh ampelopsis grossedentata leaves in the growth process is increased, the selenium content in the fresh ampelopsis grossedentata leaves is increased, and the quality of the ampelopsis grossedentata is ensured. In addition, the selenium-enriched fermentation mixed liquor obtained in the fermentation step is added with eurotium cristatum selenium-enriched and orange thermoascus aurantiacus, cyclocarya paliurus extracting solution and plantain herb extracting solution, so that the absorption of active polysaccharides, flavones, plant polyphenols, trace elements and the like in the selenium and cyclocarya paliurus extracting solution and the plantain herb extracting solution in the fermentation process of the vine tea can be effectively promoted, and the quality and the health care effect of the vine tea are improved.
Detailed Description
The above summary of the present invention is described in further detail below with reference to specific embodiments, but it should not be understood that the scope of the above subject matter of the present invention is limited to the following examples.
Introduction of some raw materials in this application:
the fresh leaves of Ampelopsis grossedentata in the examples are planted in the planting base of the large river town of the city of Laifeng county, Enshi, Hubei.
Examples sodium selenite was purchased from wuhan brilliant biotechnology limited, CAS: 10102-18-8, food grade.
In the examples, glucose was purchased from denna gmei biotechnology limited, CAS: 50-99-7, cargo number: 89-454, content: 99 percent and is in food grade.
Corn flour in the examples was purchased from shanxi snott biotechnology limited, cat #: 20201103, content: 60% and is in food grade.
In the examples citric acid was purchased from chemical Limited, QINGHAI, JUN, CAS: 77-92-9, model: 121, food grade.
In the examples, selenomethionine is purchased from Jiangsu Bangwei Biotech limited, cat #: fdqwrq, content: 99 percent and is in food grade.
In the examples, ammonium sulfate was purchased from bioengineering limited, CAS: 7783-20-2, cat no: 1236498975126, food grade.
In the examples ferrous sulfate was purchased from Hubei Hengheng science and technology Co., Ltd, CAS: 7720-78-7, food grade.
Octyl glucoside in the examples was purchased from Jiangsu Oufu Biotech, Inc., cat #: sd9+800, content of active substance: 99.9 percent.
In the examples, polyvinyl alcohol was purchased from Jiangsu Xuan Fine chemical Co., Ltd, and the molecular weight: 2 ten thousand g/mol, food grade.
In the embodiment, the eurotium cristatum is rich in selenium, is purchased from China general microbiological culture Collection center of China Committee for culture Collection of microorganisms, and has a preservation number of CGMCC No. 15395.
In the embodiment, the Thermoascus aurantiacus is purchased from China general microbiological culture Collection center with the preservation number of CGMCC 3.17992.
In the examples, the enzyme activity of the cellulase is 3.0 ten thousand U/g.
In the embodiment, the enzyme activity of the papain is 6.0 ten thousand U/g.
In the examples cyclocarya paliurus leaves were purchased from Bozhou Renyuyi Chinese medicinal materials marketing Co., Ltd.
In the examples, plantain was purchased from the Bozhou renyi Chinese medicinal materials marketing Co., Ltd and was a whole plant of plantain.
Example 1
A preparation method of vine tea comprises the following steps:
s1, picking: picking fresh leaves of Ampelopsis grossedentata, cleaning with clear water, spreading in shade, and standing for 24 hr to obtain pretreated Ampelopsis grossedentata leaves;
s2, fixation: placing the pretreated vine tea leaves in a fixation machine with the temperature of 140 ℃ and the rotation speed of 500rpm for fixation treatment until the water content of the vine tea leaves after fixation is 60 wt%;
s3, sterilization: sterilizing the deactivated folium Ampelopsis Grossdentata with 120 deg.C steam for 4 min;
s4, fermentation: soaking sterilized vine tea leaves in a selenium-rich fermentation mixed solution, wherein the mass ratio of the vine tea leaves to the selenium-rich fermentation mixed solution is 1:15, and fermenting for 5 days at a relative humidity of 65% and a temperature of 45 ℃ to obtain fermented vine tea leaves;
s5, drying: drying the fermented Ampelopsis Grossdentata leaves at 50 deg.C under 70% relative humidity, and turning the leaves once every 1 hr until the water content is 5 wt% to obtain Ampelopsis Grossdentata.
The preparation method of the selenium-rich fermentation mixed liquid comprises the following steps: uniformly mixing 4 parts by weight of sodium selenite, 10 parts by weight of glucose, 3 parts by weight of corn flour, 200 parts by weight of deionized water, 5 parts by weight of ammonium sulfate and 2 parts by weight of potassium dihydrogen phosphate to obtain a selenium-rich culture medium, sterilizing at 121 ℃ for 30min, cooling to room temperature, adding 8 wt% of zymocyte of the selenium-rich culture medium, and culturing at 30 ℃ and pH 5.0 for 20h to obtain a selenium-rich fermentation mixed solution; the zymocyte is rich in selenium by eurotium cristatum.
Example 2
A preparation method of vine tea comprises the following steps:
s1, pretreatment: controlling the growth temperature of the Ampelopsis grossedentata tea trees at 25 ℃ 28 days before the Ampelopsis grossedentata tea leaves are picked, and spraying the selenium-rich nutrient solution on the Ampelopsis grossedentata tea leaves every other day at 5 pm for one time, wherein the spraying amount is 80L per mu;
s2, picking: picking fresh Ampelopsis grossedentata leaves sprayed with selenium-rich nutrient solution, cleaning with clear water, spreading in shade, and standing for 24 hr to obtain pretreated Ampelopsis grossedentata leaves;
s3, fixation: placing the pretreated vine tea leaves in a fixation machine with the temperature of 140 ℃ and the rotation speed of 500rpm for fixation treatment until the water content of the vine tea leaves after fixation is 60 wt%;
s4, sterilization: sterilizing the deactivated folium Ampelopsis Grossdentata with 120 deg.C steam for 4 min;
s5, fermentation: soaking sterilized vine tea leaves in a selenium-rich fermentation mixed solution, wherein the mass ratio of the vine tea leaves to the selenium-rich fermentation mixed solution is 1:15, and fermenting for 5 days at a relative humidity of 65% and a temperature of 45 ℃ to obtain fermented vine tea leaves;
s6, drying: drying the fermented Ampelopsis Grossdentata leaves at 50 deg.C under 70% relative humidity, and turning the leaves once every 1 hr until the water content is 5 wt% to obtain Ampelopsis Grossdentata.
The preparation method of the selenium-rich nutrient solution comprises the following steps: mixing 4 parts by weight of sodium selenite, 2 parts by weight of ferrous sulfate, 8 parts by weight of citric acid, 15 parts by weight of monopotassium phosphate, 8 parts by weight of selenomethionine, 1 part by weight of absorption promoter and 100 parts by weight of deionized water, and stirring at room temperature and 200rpm for 3min to obtain a selenium-rich nutrient solution;
the absorption promoting agent is octyl glucoside;
the preparation method of the selenium-rich fermentation mixed liquid comprises the following steps: uniformly mixing 4 parts by weight of sodium selenite, 10 parts by weight of glucose, 3 parts by weight of corn flour, 200 parts by weight of deionized water, 5 parts by weight of ammonium sulfate and 2 parts by weight of potassium dihydrogen phosphate to obtain a selenium-rich culture medium, sterilizing at 121 ℃ for 30min, cooling to room temperature, adding 8 wt% of zymocyte of the selenium-rich culture medium, and culturing at 30 ℃ and pH 5.0 for 20h to obtain a selenium-rich fermentation mixed solution; the zymocyte is rich in selenium of eurotium cristatum.
Example 3
Essentially the same as example 2, except that: the absorption promoter is polyvinyl alcohol.
Example 4
A preparation method of vine tea comprises the following steps:
s1, pretreatment: controlling the growth temperature of the Ampelopsis grossedentata tea trees at 25 ℃ 28 days before the Ampelopsis grossedentata tea leaves are picked, and spraying the selenium-rich nutrient solution on the Ampelopsis grossedentata tea leaves every other day at 5 pm for one time, wherein the spraying amount is 80L per mu;
s2, picking: picking fresh Ampelopsis grossedentata leaves sprayed with selenium-rich nutrient solution, cleaning with clear water, spreading in shade, and standing for 24 hr to obtain pretreated Ampelopsis grossedentata leaves;
s3, fixation: placing the pretreated vine tea leaves in a fixation machine with the temperature of 140 ℃ and the rotation speed of 500rpm for fixation treatment until the water content of the vine tea leaves after fixation is 60 wt%;
s4, sterilization: sterilizing the deactivated folium Ampelopsis Grossdentata with 120 deg.C steam for 4 min;
s5, fermentation: soaking sterilized vine tea leaves in a selenium-rich fermentation mixed solution, wherein the mass ratio of the vine tea leaves to the selenium-rich fermentation mixed solution is 1:15, and fermenting for 5 days at a relative humidity of 65% and a temperature of 45 ℃ to obtain fermented vine tea leaves;
s6, drying: drying the fermented Ampelopsis Grossdentata leaves at 50 deg.C under 70% relative humidity, and turning the leaves once every 1 hr until the water content is 5 wt% to obtain Ampelopsis Grossdentata.
The preparation method of the selenium-rich nutrient solution comprises the following steps: mixing 4 parts by weight of sodium selenite, 2 parts by weight of ferrous sulfate, 8 parts by weight of citric acid, 15 parts by weight of monopotassium phosphate, 8 parts by weight of selenomethionine, 1 part by weight of absorption promoter and 100 parts by weight of deionized water, and stirring at room temperature and 200rpm for 3min to obtain a selenium-rich nutrient solution;
the absorption promoting agent is octyl glucoside;
the preparation method of the selenium-rich fermentation mixed liquid comprises the following steps: uniformly mixing 4 parts by weight of sodium selenite, 10 parts by weight of glucose, 3 parts by weight of corn flour, 200 parts by weight of deionized water, 5 parts by weight of ammonium sulfate, 2 parts by weight of potassium dihydrogen phosphate and 8 parts by weight of cyclocarya paliurus extract to obtain a selenium-rich culture medium, sterilizing at 121 ℃ for 30min, cooling to room temperature, adding 8 wt% of zymocyte of the selenium-rich culture medium, and culturing at 30 ℃ and pH 5.0 for 20h to obtain the selenium-rich fermentation mixed solution.
The zymocyte is rich in selenium of eurotium cristatum;
the preparation method of the cyclocarya paliurus extracting solution comprises the following steps: washing cyclocarya paliurus leaves with deionized water, drying in the air, crushing, sieving with a 40-mesh sieve to obtain cyclocarya paliurus leaf powder, taking 50 parts by weight of cyclocarya paliurus leaf powder, adding 100 parts by weight of deionized water and 0.5 part by weight of cellulase, adjusting the pH to 5.0, stirring at 300rpm and 35 ℃ for 5min, extracting at 45 ℃, the ultrasonic power of 150W and the ultrasonic frequency of 25kHz for 30min, filtering to obtain a primary filtrate and a primary filter residue, adding a citric acid ethanol solution with the mass of 8 wt% of the primary filter residue into the primary filter residue, stirring at 50 ℃ and 300rpm for 4h, filtering to obtain a secondary filtrate and a secondary filter residue, combining the primary filtrate and the secondary filtrate, and performing rotary evaporation and concentration under the absolute pressure of 0.05MPa and at 50 ℃ and 50rpm to 1/15 of the original mass to obtain the cyclocarya paliurus extract.
Example 5
Essentially the same as example 4, except that: the zymocyte is orange thermoascus aurantiacus.
Comparative example 1
Essentially the same as example 4, except that:
the preparation method of the selenium-rich fermentation mixed liquid comprises the following steps: uniformly mixing 4 parts by weight of sodium selenite, 10 parts by weight of glucose, 3 parts by weight of corn flour, 200 parts by weight of deionized water, 5 parts by weight of ammonium sulfate, 2 parts by weight of potassium dihydrogen phosphate and 8 parts by weight of cyclocarya paliurus extract to obtain a selenium-rich culture medium, and culturing at 30 ℃ and pH of 5.0 for 20 hours to obtain a selenium-rich fermentation mixed solution.
Comparative example 2
Essentially the same as example 4, except that:
the preparation method of the cyclocarya paliurus extracting solution comprises the following steps: washing cyclocarya paliurus leaves with deionized water, drying in the air, crushing, sieving with a 40-mesh sieve to obtain cyclocarya paliurus leaf powder, taking 50 parts by weight of cyclocarya paliurus leaf powder, adding 100 parts by weight of deionized water and 0.5 part by weight of cellulase, stirring at 300rpm and 35 ℃ for 5min, adjusting the pH to 5.0, extracting at 45 ℃, the ultrasonic power of 150W and the ultrasonic frequency of 25kHz for 30min, filtering to obtain filtrate and filter residue, and performing rotary evaporation and concentration on the filtrate under the conditions of absolute pressure of 0.05MPa, 50 ℃ and 50rpm to 1/15 of the original mass to obtain cyclocarya paliurus extract.
Example 6
Essentially the same as example 4, except that: the zymocyte is composed of eurotium cristatum selenium-rich and Thermoascus aurantiacus in a mass ratio of 3: 1.
Example 7
A preparation method of vine tea comprises the following steps:
s1, pretreatment: controlling the growth temperature of the Ampelopsis grossedentata tea trees at 25 ℃ 28 days before the Ampelopsis grossedentata tea leaves are picked, and spraying the selenium-rich nutrient solution on the Ampelopsis grossedentata tea leaves every other day at 5 pm for one time, wherein the spraying amount is 80L per mu;
s2, picking: picking fresh Ampelopsis grossedentata leaves sprayed with selenium-rich nutrient solution, cleaning with clear water, spreading in shade, and standing for 24 hr to obtain pretreated Ampelopsis grossedentata leaves;
s3, fixation: placing the pretreated vine tea leaves in a fixation machine with the temperature of 140 ℃ and the rotation speed of 500rpm for fixation treatment until the water content of the vine tea leaves after fixation is 60 wt%;
s4, sterilization: sterilizing the deactivated folium Ampelopsis Grossdentata with 120 deg.C steam for 4 min;
s5, fermentation: soaking sterilized vine tea leaves in a selenium-rich fermentation mixed solution, wherein the mass ratio of the vine tea leaves to the selenium-rich fermentation mixed solution is 1:15, and fermenting for 5 days at a relative humidity of 65% and a temperature of 45 ℃ to obtain fermented vine tea leaves;
s6, drying: drying the fermented Ampelopsis Grossdentata leaves at 50 deg.C under 70% relative humidity, and turning the leaves once every 1 hr until the water content is 5 wt% to obtain Ampelopsis Grossdentata.
The preparation method of the selenium-rich nutrient solution comprises the following steps: mixing 4 parts by weight of sodium selenite, 2 parts by weight of ferrous sulfate, 8 parts by weight of citric acid, 15 parts by weight of monopotassium phosphate, 8 parts by weight of selenomethionine, 1 part by weight of absorption promoter and 100 parts by weight of deionized water, and stirring at room temperature and 200rpm for 3min to obtain a selenium-rich nutrient solution;
the absorption promoting agent is octyl glucoside;
the preparation method of the selenium-rich fermentation mixed liquid comprises the following steps: uniformly mixing 4 parts by weight of sodium selenite, 10 parts by weight of glucose, 3 parts by weight of corn flour, 200 parts by weight of deionized water, 5 parts by weight of ammonium sulfate, 2 parts by weight of potassium dihydrogen phosphate, 5 parts by weight of cyclocarya paliurus extract and 3 parts by weight of plantain extract to obtain a selenium-rich culture medium, sterilizing at 121 ℃ for 30min, cooling to room temperature, adding 8 wt% of zymocyte of the selenium-rich culture medium, and culturing at 30 ℃ and pH 5.0 for 20h to obtain the selenium-rich fermentation mixed solution.
The fermentation bacteria consist of eurotium cristatum selenium-rich and Thermoascus aurantiacus in a mass ratio of 3: 1;
the preparation method of the cyclocarya paliurus extracting solution comprises the following steps: washing cyclocarya paliurus leaves with deionized water, drying in the air, crushing, sieving with a 40-mesh sieve to obtain cyclocarya paliurus leaf powder, taking 50 parts by weight of cyclocarya paliurus leaf powder, adding 100 parts by weight of deionized water and 0.5 part by weight of cellulase, adjusting the pH to 5.0, stirring at 300rpm and 35 ℃ for 5min, extracting at 45 ℃, the ultrasonic power of 150W and the ultrasonic frequency of 25kHz for 30min, filtering to obtain a primary filtrate and a primary filter residue, adding a citric acid ethanol solution with the mass of 8 wt% of the primary filter residue into the primary filter residue, stirring at 50 ℃ and 300rpm for 4h, filtering to obtain a secondary filtrate and a secondary filter residue, combining the primary filtrate and the secondary filtrate, and performing rotary evaporation and concentration under the absolute pressure of 0.05MPa and at 50 ℃ and 50rpm to 1/15 of the original mass to obtain the cyclocarya paliurus extract.
The preparation method of the plantain herb extracting solution comprises the following steps: cleaning herba plantaginis, air drying, pulverizing, and sieving with 60 mesh sieve to obtain herba plantaginis powder; adding 55 parts by weight of plantain powder into 95 parts by weight of deionized water, uniformly mixing, and performing microwave treatment for 5min under the condition of 280W; cooling to room temperature, adding 0.4 weight part of cellulase and 0.2 weight part of papain, uniformly mixing, adjusting the pH to 5.0, stirring at 80rpm and 45 ℃ for 25min, filtering to obtain a filtrate I and a residue I, adding 65 wt% of ethanol aqueous solution into the residue I, carrying out reflux extraction at 86 ℃ for 200min, wherein the mass ratio of the residue I to the ethanol aqueous solution is 1:5, filtering to obtain a filtrate II and a residue II, combining the filtrate I and the filtrate II, and carrying out rotary evaporation concentration under the conditions of absolute pressure of 0.05MPa, 50 ℃ and 50rpm to 1/10 of the original mass to obtain the plantain herb extracting solution. The ampelopsis grossedentata of example 7 was subjected to a blood lipid lowering effect test in accordance with the method of test example 3, and the triglyceride lowering range was 29.3% and the cholesterol lowering range was 23.9%.
Comparative example 3
A preparation method of vine tea comprises the following steps:
s1, pretreatment: controlling the growth temperature of the Ampelopsis grossedentata tea trees at 25 ℃ 28 days before the Ampelopsis grossedentata tea leaves are picked, and spraying the selenium-rich nutrient solution on the Ampelopsis grossedentata tea leaves every other day at 5 pm for one time, wherein the spraying amount is 80L per mu;
s2, picking: picking fresh Ampelopsis grossedentata leaves sprayed with selenium-rich nutrient solution, cleaning with clear water, spreading in shade, and standing for 24 hr to obtain pretreated Ampelopsis grossedentata leaves;
s3, fixation: placing the pretreated vine tea leaves in a fixation machine with the temperature of 140 ℃ and the rotation speed of 500rpm for fixation treatment until the water content of the vine tea leaves after fixation is 60 wt%;
s4, sterilization: sterilizing the deactivated folium Ampelopsis Grossdentata with 120 deg.C steam for 4 min;
s5, fermentation: soaking sterilized vine tea leaves in a selenium-rich fermentation mixed solution, wherein the mass ratio of the vine tea leaves to the selenium-rich fermentation mixed solution is 1:15, and fermenting for 5 days at a relative humidity of 65% and a temperature of 45 ℃ to obtain fermented vine tea leaves;
s6, drying: drying the fermented Ampelopsis Grossdentata leaves at 50 deg.C under 70% relative humidity, and turning the leaves once every 1 hr until the water content is 5 wt% to obtain Ampelopsis Grossdentata.
The preparation method of the selenium-rich nutrient solution comprises the following steps: mixing 4 parts by weight of sodium selenite, 2 parts by weight of ferrous sulfate, 8 parts by weight of citric acid, 15 parts by weight of monopotassium phosphate, 8 parts by weight of selenomethionine, 1 part by weight of absorption promoter and 100 parts by weight of deionized water, and stirring at room temperature and 200rpm for 3min to obtain a selenium-rich nutrient solution;
the absorption promoting agent is octyl glucoside;
the preparation method of the selenium-rich fermentation mixed liquid comprises the following steps: uniformly mixing 4 parts by weight of sodium selenite, 10 parts by weight of glucose, 3 parts by weight of corn flour, 200 parts by weight of deionized water, 5 parts by weight of ammonium sulfate, 2 parts by weight of monopotassium phosphate and 8 parts by weight of plantain herb extracting solution to obtain a selenium-rich culture medium, sterilizing at 121 ℃ for 30min, cooling to room temperature, adding 8 wt% of zymogen of the selenium-rich culture medium, and culturing at 30 ℃ and pH 5.0 for 20h to obtain the selenium-rich fermentation mixed solution.
The fermentation bacteria consist of eurotium cristatum selenium-rich and Thermoascus aurantiacus in a mass ratio of 3: 1;
the preparation method of the plantain herb extracting solution comprises the following steps: cleaning herba plantaginis, air drying, pulverizing, and sieving with 60 mesh sieve to obtain herba plantaginis powder; adding 55 parts by weight of plantain powder into 95 parts by weight of deionized water, uniformly mixing, and performing microwave treatment for 5min under the condition of 280W; cooling to room temperature, adding 0.4 weight part of cellulase and 0.2 weight part of papain, uniformly mixing, adjusting the pH to 5.0, stirring at 80rpm and 45 ℃ for 25min, filtering to obtain a filtrate I and a residue I, adding 65 wt% of ethanol aqueous solution into the residue I, carrying out reflux extraction at 86 ℃ for 200min, wherein the mass ratio of the residue I to the ethanol aqueous solution is 1:5, filtering to obtain a filtrate II and a residue II, combining the filtrate I and the filtrate II, and carrying out rotary evaporation concentration under the conditions of absolute pressure of 0.05MPa, 50 ℃ and 50rpm to 1/10 of the original mass to obtain the plantain herb extracting solution. The ampelopsis grossedentata of comparative example 3 was tested for its blood lipid-lowering effect by referring to the method of test example 3, and its triglyceride-lowering range was 28.0% and cholesterol-lowering range was 21.8%.
Test example 1
Determining the total selenium content of the vine tea: referring to 'evaluation of leaching rate and effectiveness of selenium in selenium-rich tea' of Chengyong Bo et al, 1g of the ampelopsis grossedentata samples in examples 1-6 and comparative examples 1-2 were weighed into a small beaker, and leached with boiling ultrapure water for 3 times, 100ml of water was added each time, the leaching time was 10min, 2 times of parallel leaching were carried out, 3 times of leaching liquor were combined, and the total selenium content in the ampelopsis grossedentata was determined. The results are shown in Table 1.
TABLE 1 Total selenium content determination of Ampelopsis grossedentata
From the above results, it can be seen that, compared with examples 1-2 and 2, the selenium-rich nutrient solution is sprayed on the growing ampelopsis grossedentata leaves before spraying and picking, so that the absorption of organic selenium can be well promoted, compared with examples 2-3, the absorption promoting agent added in the selenium-rich nutrient solution is different from the absorption promoting agent added in the selenium-rich nutrient solution, and the octyl glucoside adopted in example 2 as the absorption promoting agent can not only wet the surface of the ampelopsis grossedentata leaves, but also play a role in permeation, when ferrous sulfate increases the permeability of cells, the capillary pores on the leaves can be opened, so that the absorption of the selenium-rich nutrient solution is further promoted, and the content of organic selenium in the fresh leaves of the ampelopsis grossedentata vines is increased; comparing example 4 with comparative example 1, in example 4, fermentation bacteria are added to the selenium-rich fermentation mixed solution, so that the conversion rate and the absorption rate of organic selenium can be promoted, and the selenium content can be improved by adding the cyclocarya paliurus extract in the fermentation process, because the cyclocarya paliurus extract also contains selenium element and is absorbed by the vine tea leaves in the fermentation process, the total selenium content of the vine tea is increased; finally, comparing examples 4-6, example 6 is that the Eurotium cristatum selenium-rich and the Thermoascus aurantiacus are adopted as zymocyte to play a role in synergy, so that the transformation and absorption of selenium are promoted together, the two synergies are used for further improving the content of organic selenium in the vine tea.
Test example 2
Determination of DPPH radical scavenging activity: the method adopts a colorimetric method for determination, and the determination principle is as follows: the DPPH free gene has a single electron, so that the DPPH free gene has extremely strong absorption (dark purple) at 525nm, when a free radical scavenger exists, the absorption is gradually disappeared due to the pairing with the single electron, the fading degree is in quantitative relation with the number of the received electrons, and therefore, the DPPH free gene can be quantitatively analyzed by a spectrophotometry method. The higher the scavenging rate of the antioxidant for scavenging free radicals, the more antioxidant it is.
(1) Sample preparation: weighing 1g of the vine tea obtained in the embodiments 2-6 and the comparative example 2, adding 30mL of deionized water at 98 ℃, leaching for 30min in a water bath at 98 ℃, filtering with 300-mesh filter cloth while hot, and fixing the volume to 50mL to obtain the vine tea.
(2) Reagent: 0.1mM DPPH-methanol solution, 0.05mol/L Tris-HCl buffer.
(3) The method comprises the following operation steps: sample tube: 2mL of DPPH-methanol solution, 0.9mL of Tris-HCl buffer and 0.5mL of sample. And (4) control: 2mL of DPPH-methanol solution, 0.9mL of buffer, and 0.5mL of methanol. And (5) zero setting of the distilled water pipe. After adding the sample, the reaction was carried out in the dark for 30min, and the absorbance was measured at 517 nm.
Calculating the formula: clearance (%) - (control tube absorbance-sample tube absorbance)/control tube absorbance × 100. Groups 4 were tested in parallel and averaged, and the results are shown in table 2.
TABLE 2 measurement results of DPPH radical scavenging activity
| DPPH radical scavenging ratio (%) | |
| Example 2 | 60.7 |
| Example 4 | 67.3 |
| Example 5 | 66.5 |
| Example 6 | 69.7 |
| Comparative example 2 | 64.2 |
From the above results, it can be seen that, the active ingredients dihydromyricetin in the vine tea and the active substances in the cyclocarya paliurus extract, such as phenols, terpenoids, polysaccharides, etc., have a strong radical scavenging effect, and the vine tea itself also has a strong radical scavenging function, specifically, comparative example 2 and example 4 show that the DPPH radical scavenging rate of the cyclocarya paliurus extract added in example 4 is significantly higher than the DPPH radical scavenging rate of the cyclocarya paliurus extract not added in example 2, the reason for this is simple, the active substances in the cyclocarya paliurus extract, such as phenols, terpenoids, polysaccharides, etc., have a strong radical scavenging effect, the vine tea leaves absorb the extractive solution of the vine liquid during the fermentation process, the active ingredients with radical scavenging effect are leached from the extractive solution of the vine tea brewed with boiling water, comparative example 4 and comparative example 2, and comparative example 2 do not adopt secondary extraction during the extractive solution of the vine liquid, the content of active ingredients in the green cash liquid extracting solution is reduced, and the content of the active ingredients in the green cash liquid extracting solution absorbed by the vine tea is correspondingly reduced during fermentation, so that the capacity of removing free radicals is also reduced.
Test example 3
And (3) testing the blood fat reducing effect: weighing 3g of Ampelopsis grossedentata prepared in examples 2-6 and comparative examples 1-2, soaking in 100mL of 90 deg.C water for 10min, filtering to remove tea, extracting for 3 times, retaining the filtrate for 3 times, and concentrating the filtrate to a volume of 5mL to obtain Ampelopsis grossedentata concentrated solution. The hyperlipidemic rats are divided into 1 control group and 3 experimental groups, each group comprises 30 rats, each group is half male and female, and the weight of the hyperlipidemic rats in each group is 150 +/-2 g. The experimental group respectively gavage the ampelopsis grossedentata concentrated solution of the examples 4-5 and the comparative example 2 according to the dose of 3g/kg for rats with 12 hours in the open abdomen, and the control group continuously gavage tail blood after 30 days of continuous gavage according to the dose of 3g/kg, and then the cholesterol and triglyceride content in the serum is measured. The experimental group and the control group were kept under normal conditions except for gavage, and the rats were allowed to freely take water and food. The contents of cholesterol and triglyceride in the serum of the experimental group and the control group after 30 days of gastric lavage are compared, and the reduction amplitude is calculated. The cholesterol reduction amplitude is equal to (average cholesterol content in control group-average cholesterol content in experimental group)/average cholesterol content in control group is multiplied by 100%. The triglyceride reduction amplitude is (average content of triglyceride in control group-average content of triglyceride in experimental group)/average content of triglyceride in control group x 100%. The test results are shown in Table 3.
TABLE 3 test results of blood lipid lowering effect
| Triglyceride reduction amplitude (%) | Cholesterol reduction amplitude (%) | |
| Example 2 | 16.9 | 17.8 |
| Example 4 | 26.4 | 20.8 |
| Example 5 | 27.9 | 21.6 |
| Example 6 | 28.9 | 23.1 |
| Comparative example 1 | 20.4 | 18.5 |
| Comparative example 2 | 24.2 | 18.8 |
From the above results, it can be seen that the flavone, which is an active ingredient in the vine tea, can remove harmful ketone bodies from human bodies and remove blood stains in blood, so as to achieve the effect of regulating blood fat, and the polysaccharide in the cyclocarya paliurus extract has a good function of reducing blood fat.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (7)
1. The preparation method of the vine tea is characterized by comprising the following steps:
s1, pretreatment: controlling the growth temperature of the Ampelopsis grossedentata tea trees to be 20-30 ℃ 25-30 days before the Ampelopsis grossedentata tea leaves are picked, and spraying the selenium-rich nutrient solution on the Ampelopsis grossedentata tea leaves at 4-6 pm every other day, wherein the spraying amount is 60-120L per mu;
s2, picking: picking fresh Ampelopsis grossedentata leaves sprayed with selenium-rich nutrient solution, cleaning with clear water, spreading in shade, and standing for 20-40 hr to obtain pretreated Ampelopsis grossedentata leaves;
s3, fixation: placing the pretreated vine tea leaves in a water-removing machine with the temperature of 130-;
s4, sterilization: sterilizing the deactivated vine tea leaves for 2-5min by steam at the temperature of 100-;
s5, fermentation: soaking sterilized vine tea leaves in a selenium-rich fermentation mixed solution, wherein the mass ratio of the vine tea leaves to the selenium-rich fermentation mixed solution is 1 (10-25), and fermenting for 2-8 days at a relative humidity of 60-70% and a temperature of 30-55 ℃ to obtain fermented vine tea leaves; the preparation method of the selenium-rich fermentation mixed liquid comprises the following steps: uniformly mixing 2-5 parts by weight of sodium selenite, 5-15 parts by weight of glucose, 1-6 parts by weight of corn flour, 400 parts by weight of 100-plus materials of water, 2-6 parts by weight of ammonium sulfate, 1-3 parts by weight of potassium dihydrogen phosphate, 5-10 parts by weight of cyclocarya paliurus extract and 1-5 parts by weight of plantain herb extract, sterilizing at the temperature of 100-plus materials of 130 ℃ for 20-50min, cooling to room temperature to obtain a selenium-rich culture medium, adding 5-10 wt% of zymophyte of the selenium-rich culture medium, and culturing at the temperature of 25-35 ℃ and the pH of 4.0-7.0 for 16-24h to obtain a selenium-rich fermentation mixed solution;
s6, drying: drying the fermented Ampelopsis Grossdentata leaves at 35-55 deg.C under relative humidity of 60-80%, turning over the leaves every 0.5-1.5 hr until the water content is 3-7 wt% to obtain Ampelopsis Grossdentata tea.
2. The method for preparing ampelopsis grossedentata according to claim 1, wherein the method for preparing the extract of plantain comprises the steps of: cleaning herba plantaginis, air drying, pulverizing, and sieving with 20-80 mesh sieve to obtain herba plantaginis powder; adding 50-60 parts by weight of plantain powder into 80-100 parts by weight of water, uniformly mixing, and performing microwave treatment for 2-6min under the conditions of 200-300W; cooling to room temperature, adding 0.2-0.6 weight part of cellulase and 0.1-0.3 weight part of papain, uniformly mixing, adjusting the pH to 5-7, stirring at 50-100rpm at 40-50 ℃ for 20-30min, filtering to obtain a filtrate I and a filter residue I, adding 60-70 wt% of ethanol aqueous solution into the filter residue I, carrying out reflux extraction at 80-90 ℃ for 150-250min, wherein the mass ratio of the filter residue I to the ethanol aqueous solution is 1 (2-7), filtering to obtain a filtrate II and a filter residue II, combining the filtrate I and the filtrate II, and carrying out rotary evaporation and concentration to 1/10-1/5 of the original mass to obtain the plantain herb extract.
3. The preparation method of the ampelopsis grossedentata according to claim 1, wherein the preparation method of the cyclocarya paliurus extract comprises the following steps: washing cyclocarya paliurus leaves with water, drying in the air, crushing, sieving with a 30-60-mesh sieve to obtain cyclocarya paliurus leaf powder, taking 40-60 parts by weight of cyclocarya paliurus leaf powder, adding 80-150 parts by weight of water and 0.1-1 part by weight of cellulase, adjusting the pH to 4.5-6, stirring at 200-40 ℃ and 500rpm for 3-10min, extracting at 40-50 ℃ and ultrasonic frequency of 20-40kHz for 20-50min, filtering to obtain a primary filtrate and a primary filter residue, adding a citric acid ethanol solution with the mass of 5-15 wt% of the primary filter residue into the primary filter residue, stirring at 45-55 ℃ and 200-400rpm for 3-5h, filtering to obtain a secondary filtrate and a secondary filter residue, merging the primary filtrate and the secondary filtrate, concentrating by rotary evaporation to 1/20-1/10 of the original mass to obtain cyclocarya paliurus extract.
4. A method of preparing ampelopsis grossedentata according to claim 1, wherein said selenium-enriched nutrient solution of step S1 is prepared by: mixing 2-7 parts by weight of sodium selenite, 1-3 parts by weight of ferrous sulfate, 5-18 parts by weight of citric acid, 10-20 parts by weight of monopotassium phosphate, 5-12 parts by weight of selenomethionine, 0.5-2 parts by weight of absorption promoter and 80-120 parts by weight of water, and stirring at 100-300rpm at room temperature for 2-5min to obtain the selenium-rich nutrient solution.
5. A process for preparing Ampelopsis grossedentata according to claim 4, wherein said absorption promoter is octyl glucoside or polyvinyl alcohol.
6. The method for preparing Ampelopsis grossedentata according to claim 1, wherein said fermentation tubes are Eurotium cristatum selenium-enriched and/or Thermoascus aurantiacus.
7. Vine tea, characterized in that it is prepared by the process according to any one of claims 1 to 6.
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