阅读说明：本技术 一种青钱柳金桔红茶菌益生饮料的制备 (Preparation method of cyclocarya paliurus, kumquat and black tea fungus probiotic beverage ) 是由 申椰雯 朱孔飞 田方 金俊 岳孟雪 于 2021-02-05 设计创作，主要内容包括：本发明提供了一种以青钱柳茶叶、金桔原汁为原料,通过红茶菌发酵制备一款青钱柳金桔红茶菌益生饮料,该青钱柳金桔红茶菌益生饮料具有降三高的功效,能有效降低人体内的甘油三酯和胆固醇。与传统红茶菌饮料或青钱柳茶相比,功效更强、口感更佳、营养价值更高。(The invention provides a cyclocarya paliurus tea and kumquat raw juice probiotic beverage prepared by fermenting black tea bacteria, which has the efficacy of reducing high blood pressure, high blood sugar and high blood fat, and can effectively reduce triglyceride and cholesterol in human bodies. Compared with the traditional black tea fungus beverage or cyclocarya paliurus tea, the black tea fungus beverage has stronger efficacy, better taste and higher nutritional value.)

1. A preparation method of cyclocarya paliurus, kumquat and black tea fungus probiotic beverage is characterized by comprising the following steps: the cyclocarya paliurus and kumquat black tea fungus probiotic beverage is prepared by taking cyclocarya paliurus tea water and kumquat raw juice as raw materials, taking cane sugar as a carbon source and inoculating black tea fungus mother liquor.

2. The preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage according to claim 1, wherein the preparation method comprises the following steps: the preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage specifically comprises the following steps:

1) preparing cyclocarya paliurus tea: weighing cyclocarya paliurus tea leaves, brewing the cyclocarya paliurus tea leaves in boiling water for 10-15 min to prepare tea water, and filtering to obtain tea water filtrate;

2) preparing a black tea fungus mother solution: adding sucrose into the tea water filtrate, stirring for dissolving, cooling, adding black tea fungus starter, standing at 25 deg.C, and ventilating to culture until film appears on the surface to obtain black tea fungus mother liquor;

3) preparation of kumquat raw juice

a. Selecting raw materials: the cumquats are required to have the maturity of about ninety-five, golden yellow, strong fragrance, crisp texture, no scab and no rot, and the impurities such as fruit stalks and the like are removed and cleaned for standby;

b. blanching, softening and debitterizing: heating to 90-95 ℃ by introducing steam into a stainless steel jacketed kettle containing two thirds of water, then adding the cleaned kumquats, and blanching for 2-3 min; soaking in saturated limewater for 3.5 h;

c. pulping: cleaning the soaked kumquats with purified water, adding purified water according to the proportion of 1:2, placing the kumquats in a beater to be beaten, and separating pulp through a stainless steel screen with the aperture of 100 meshes to obtain filtrate;

d. homogenizing: adding 0.4% of beta-cyclodextrin and 0.02% of triglycerol monoglyceride, preheating the juice solution to 50-60 ℃, and homogenizing by a high-pressure homogenizer to further micronize particles;

e. sterilization and cooling: sterilizing the mixture at 85 deg.C for 15-20 min, and rapidly cooling in cold water to about 35 deg.C to obtain kumquat juice;

4) fermentation of

Firstly, putting sucrose into a clean glass ware, pouring a proper amount of boiling water, stirring and dissolving, and transferring to a volumetric flask to fix the volume to prepare a 10% sucrose aqueous solution; adding kumquat juice, cooling, inoculating black tea fungus mother liquor, ventilating, standing, fermenting and culturing at 20-30 ℃, and stopping fermentation when the pH value of the fermentation solution is 3.5-4.5;

5) blending: stevioside is added into the fermentation liquor for seasoning, most of sugar in the fermented kumquat and black tea fungus composite fermentation liquor is consumed by lactic acid bacteria, and natural sweet substances are selected for blending the beverage in order to improve the flavor;

6) and (3) sterilization: and (3) instantly sterilizing at the high temperature of 90-95 ℃ for 10-15 s to meet the sterilization requirement, then carrying out aseptic hot filling, and cooling to 37-38 ℃ to obtain the cyclocarya paliurus kumquat and black tea fungus probiotic beverage.

3. The preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage according to claim 2, wherein the preparation method comprises the following steps: the weight-volume ratio of the cyclocarya paliurus tea leaves to the boiling water in the step 1) is 1:100, and the unit is g/ml.

4. The preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage according to claim 2, wherein the preparation method comprises the following steps: the weight volume ratio of the sucrose to the tea water filtrate in the step 2) is 1:10 unit g/ml.

5. The preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage according to claim 2, wherein the preparation method comprises the following steps: the black tea fungus starter in the step 2) comprises acetic acid bacteria, lactic acid bacteria and saccharomycetes.

6. The preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage according to claim 5, wherein the preparation method comprises the following steps: the content of each strain in the black tea fungus starter in the step 2) is calculated by the total number of colonies, and the unit cfu/mL: acetic acid bacteria 2.0 × 10⁶～6.0×10⁶(ii) a Lactobacillus 2.0X 10⁶～6.0×10⁶(ii) a Yeast 1.0 × 10⁶～3.0×10⁶。

7. The preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage according to claim 2, wherein the preparation method comprises the following steps: the volume ratio of the black tea fungus starter to the tea water filtrate in the step 2) is 1: 20.

8. The preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage according to claim 2, wherein the preparation method comprises the following steps: the volume percentages of the 10% sucrose aqueous solution, the kumquat raw juice and the black tea fungus mother liquor in the step 4) are as follows:

10 to 20 percent of kumquat raw juice

1-5% of 10% sucrose aqueous solution

75-89% of black tea fungus mother liquor.

9. The preparation method of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage according to claim 2, wherein the preparation method comprises the following steps: and in the step 5), stevioside is added into the fermentation liquor for seasoning, and the stevioside is added to adjust the sugar degree of the beverage to be 18 degrees Bx.

Technical Field

The invention relates to the technical field of food, and in particular relates to a preparation method of a cyclocarya paliurus kumquat and black tea fungus probiotic beverage.

Background

The black tea fungus fermented drink is a folk traditional acidic drink with a long history. Generally, the black tea sugar water is used as a raw material and is fermented together by a plurality of microorganisms such as acetic acid bacteria, saccharomycetes, lactic acid bacteria and the like. The fermented beverage is rich in nutritional ingredients such as lactic acid, acetic acid, yeast amino acid, vitamins, lecithin, mineral substances and the like, has various probiotic and health-care effects of improving intestinal functions, resisting tumors, delaying senescence, enhancing mouthfeel and the like, is sour, sweet and delicious, is cool and antipyretic, and has unique flavor. At present, few probiotic fermented tea beverage products are sold in the market, and the research on the cyclocarya paliurus and black tea fungus fermented beverage is nearly blank. Most of the cyclocarya paliurus tea is brewed and drunk as tea water, and the taste of the cyclocarya paliurus tea is bitter and astringent, so that the palatability of the product is poor. The black tea fungus fermented beverage is early in origin in China, but has single taste, lacks deep processing, is difficult to adapt to the innovative and diversified demands of consumers on the beverage and is difficult to be pushed to a wider market.

Disclosure of Invention

The invention provides a cyclocarya paliurus tea and kumquat raw juice probiotic beverage prepared by fermenting black tea bacteria, which has the efficacy of reducing high blood pressure, high blood sugar and high blood fat, and can effectively reduce triglyceride and cholesterol in human bodies. Compared with the traditional black tea fungus beverage or cyclocarya paliurus tea, the black tea fungus beverage has stronger efficacy, better taste and higher nutritional value.

A preparation method of cyclocarya paliurus and kumquat black tea fungus probiotic beverage comprises taking cyclocarya paliurus tea water and kumquat raw juice as raw materials, taking sucrose as a carbon source, and inoculating black tea fungus mother liquor.

Further, the preparation of the cyclocarya paliurus kumquat and black tea fungus probiotic beverage specifically comprises the following steps:

1) preparing cyclocarya paliurus tea: weighing cyclocarya paliurus tea leaves, brewing the cyclocarya paliurus tea leaves in boiling water for 10-15 min to prepare tea water, and filtering to obtain tea water filtrate;

2) preparing a black tea fungus mother solution: adding sucrose into the tea water filtrate, stirring for dissolving, cooling, adding black tea fungus starter, standing at 25 deg.C, and ventilating to culture until film appears on the surface to obtain black tea fungus mother liquor;

3) preparation of kumquat raw juice

a. Selecting raw materials: the cumquats are required to have the maturity of about ninety-five, golden yellow, strong fragrance, crisp texture, no scab and no rot, and the impurities such as fruit stalks and the like are removed and cleaned for standby;

b. blanching, softening and debitterizing: heating to 90-95 ℃ by introducing steam into a stainless steel jacketed kettle containing two thirds of water, then adding the cleaned kumquats, and blanching for 2-3 min; taking out after blanching, and putting into saturated limewater to soak for 3.5 h;

c. pulping: cleaning the soaked kumquats with purified water, adding the purified water according to the weight ratio of 1:2, placing the kumquats in a beater to be beaten, and separating pulp through a stainless steel screen mesh with the aperture of 100 meshes to obtain filtrate;

d. homogenizing: adding 0.4% of beta-cyclodextrin and 0.02% of triglycerol monoglyceride, preheating the juice solution to 50-60 ℃, and homogenizing by a high-pressure homogenizer to further micronize particles;

e. sterilization and cooling: sterilizing the mixture at 85 deg.C for 15-20 min, and rapidly cooling in cold water to about 35 deg.C to obtain kumquat juice;

4) fermentation: firstly, putting sucrose into a clean glass ware, pouring a proper amount of boiling water, stirring and dissolving, and transferring to a volumetric flask to fix the volume to prepare a 10% sucrose aqueous solution; adding kumquat juice, cooling, inoculating black tea fungus mother liquor, ventilating, standing, fermenting and culturing at 20-30 ℃, and stopping fermentation when the pH value of the fermentation solution is 3.5-4.5;

5) blending: stevioside is added into the fermentation liquor for seasoning, most of sugar in the fermented kumquat and black tea fungus composite fermentation liquor is consumed by lactic acid bacteria, and natural sweet substances are selected for blending the beverage in order to improve the flavor;

6) and (3) sterilization: and (3) instantly sterilizing at the high temperature of 90-95 ℃ for 10-15 s to meet the sterilization requirement, then carrying out aseptic hot filling, and cooling to 37-38 ℃ to obtain the cyclocarya paliurus kumquat and black tea fungus probiotic beverage.

Preferably, the weight-volume ratio of the cyclocarya paliurus tea leaves to the boiling water in the step 1) is 1:100, and the unit is g/ml.

Preferably, the weight-volume ratio of the sucrose to the tea water filtrate in the step 2) is 1:10, and the unit is g/ml.

Preferably, the black tea fungus starter in the step 2) comprises acetic acid bacteria, lactic acid bacteria and yeast.

Further preferably, the content of each strain in the black tea fungus starter in the step 2) is calculated by the total number of colonies (cfu/mL): acetic acid bacteria 2.0 × 10⁶～6.0×10⁶(ii) a Lactobacillus 2.0X 10⁶～6.0×10⁶(ii) a Yeast 1.0 × 10⁶～3.0×10⁶。

Preferably, the volume ratio of the black tea fungus starter to the tea water filtrate in the step 2) is 1: 20.

Preferably, the 10% sucrose aqueous solution, the kumquat raw juice and the black tea fungus mother liquor in the step 4) have the following volume percentages:

10 to 20 percent of kumquat raw juice

1-5% of 10% sucrose aqueous solution

75 to 89 percent of black tea fungus mother liquor

Preferably, stevioside serving as a 'natural' raw material is added into the fermentation liquor in the step 5) to replace the traditional cane sugar for seasoning, and the stevioside is added to adjust the sugar degree of the beverage to be 18 degrees Bx.

The cyclocarya paliurus and kumquat black tea fungus probiotic beverage is prepared by taking cyclocarya paliurus tea as a matrix, fermenting the cyclocarya paliurus tea by using black tea fungus, enriching kumquat and harmonizing flavor and utilizing a probiotic composite fermentation method. Compared with a single black tea fungus fermented beverage or cyclocarya paliurus tea, the product is novel and unique in taste, rich in nutrition and diversified in health care effect.

A. The product has the characteristic of low caffeine content and multiple health care effects. The traditional black tea and green tea contain certain content of purine alkaloids such as caffeine, theobromine, theophylline and the like, the cyclocarya paliurus tea contains low content of the alkaloids, and the cyclocarya paliurus leaves contain various functional substances (saccharides, flavones, terpenes, proteins, mineral substances, phenolic acids, vitamins, saponins and the like), so that the effects of resisting oxidation, reducing blood sugar, resisting tumors, resisting bacteria, improving immunity and the like are achieved.

B. The product is rich in various probiotic fermentation products. The black tea fungus contains three probiotics beneficial to human bodies: yeast, acetic acid bacteria and lactic acid bacteria. The probiotic bacteria can convert cyclocarya paliurus macromolecular polysaccharide into oligomeric polysaccharide in the fermentation process, and the produced abundant organic acids and esters can endow the beverage with good flavor, and meanwhile, the fermentation liquor contains metabolites such as vitamins, amino acids, tea polyphenol and the like, so that the beverage has various health-care functions of promoting digestion, regulating organism immunity, inhibiting bacteria, resisting oxidation, preventing cardiovascular diseases and the like.

C. The product has unique and innovative flavor. The kumquat juice can provide a certain carbon source for probiotic fermentation together with substances such as polysaccharide in the cyclocarya paliurus, so that the use of cane sugar is reduced, and the original bitter taste of the cyclocarya paliurus tea can be neutralized or masked, so that a new flavor is created. Meanwhile, the product is rich in vitamin C, kumquat glycoside and other nutritional ingredients in kumquats, and the antioxidant effect of the product is improved.

D. The product has low sugar and low calorie. The safe and healthy food becomes the mainstream appeal of modern consumers, the product uses natural sweet substances to replace cane sugar, the consumption of sugar is reduced again on the basis of ensuring the taste of the beverage, and the aim is to create a low-sugar and low-calorie healthy beverage.

Detailed Description

The following examples are intended to further illustrate the present invention, but they are not intended to limit or restrict the scope of the invention. The solvent and apparatus used in the present invention are not particularly limited, and commercially available conventional solvents and apparatuses can be used.

Example 1 preparation of kumquat raw juice

Selecting clean and non-rotten kumquats with the maturity of about ninety-five and golden yellow, removing 800g of fruit stalks and other impurities, cleaning, putting the kumquats into a stainless steel jacketed pan filled with two thirds of water, introducing steam to heat the kumquats to 95 ℃, then adding the cleaned kumquats, and blanching for 2 min; taking out, soaking in saturated limewater for about 3.5h, cleaning with purified water, adding purified water at a weight ratio of 1:2, pulping in a pulping machine, separating with 100 mesh stainless steel sieve, and removing seed, skin, and coarse fiber; adding 0.4% (weight ratio of beta-cyclodextrin to filtrate) of beta-cyclodextrin and 0.02% (weight ratio of triglycerol monoglyceride to filtrate) of triglycerol monoglyceride into the filtrate, preheating the juice solution to 55 deg.C before homogenizing, homogenizing with a high pressure homogenizer, sterilizing at 85 deg.C for 20min, and rapidly cooling to about 35 deg.C in cold water.

Example 2 cyclocarya paliurus kumquat and black tea fungus probiotic beverage preparation

1) Preparing cyclocarya paliurus tea: weighing 60g parts of cyclocarya paliurus tea, soaking in 6000ml of boiling water for 15min to prepare tea water, and filtering to obtain tea water filtrate;

2) preparing a black tea fungus mother solution: adding 300g sucrose into 3000ml tea water filtrate obtained in step 1), stirring for dissolving, cooling, adding 150ml black tea fungus starter (acetic acid bacteria 4.0 × 10)⁶cfu/mL; lactic acid bacteria 4.0X 10⁶cfu/mL; yeast 2.0 × 10⁶cfu/mL), standing and ventilating at 25 ℃ until a film appears on the surface, namely the black tea fungus mother liquor, for later use;

3) preparation of kumquat raw juice

The kumquat raw juice prepared in example 1 was used directly.

4) Fermentation: firstly, putting sucrose into a clean glass ware, pouring a proper amount of boiling water, stirring and dissolving, and then transferring to a volumetric flask to fix the volume to prepare a 10% sucrose syrup aqueous solution; adding kumquat juice, cooling, and inoculating black tea fungus mother liquor, wherein the volume of each component is as follows:

250ml of kumquat raw juice

50ml of 10% sucrose aqueous solution

Black tea fungus mother liquor 1700ml

Ventilating, standing, fermenting and culturing at 20 ℃, and stopping fermentation when the pH of the fermentation solution is 3.5-4.5;

5) blending: adding stevioside serving as a 'natural' raw material into the fermentation liquor to replace the traditional cane sugar for seasoning until the sugar degree is 18 degrees Bx;

6) and (3) sterilization: the sterilizing requirement can be met by instantly sterilizing at the high temperature of 95 ℃ for 15s, then carrying out aseptic hot filling, and cooling to 38 ℃ to obtain the cyclocarya paliurus kumquat black tea fungus probiotic beverage.

Example 3 cyclocarya paliurus kumquat and black tea fungus probiotic beverage preparation

1) Preparing cyclocarya paliurus tea: weighing 50g parts of cyclocarya paliurus tea, soaking in 5000ml of boiling water for 12min to obtain tea water, and filtering to obtain tea water filtrate;

2) preparing a black tea fungus mother solution: adding 300g sucrose into 3000ml tea water filtrate obtained in step 1), stirring for dissolving, cooling, adding 150ml black tea fungus starter (acetic acid bacteria 4.0 × 10)⁶cfu/mL; lactic acidBacterium 4.0X 10⁶cfu/mL; yeast 2.0 × 10⁶cfu/mL), standing and ventilating at 25 ℃ until a film appears on the surface, namely the black tea fungus mother liquor, for later use;

3) preparation of kumquat raw juice

The kumquat raw juice prepared in example 1 was used directly.

4) Fermentation: firstly, putting sucrose into a clean glass ware, pouring a proper amount of boiling water, stirring and dissolving, and then transferring to a volumetric flask to fix the volume to prepare a 10% sucrose syrup aqueous solution; adding kumquat juice, cooling, and inoculating black tea fungus mother liquor, wherein the added content of each component is as follows:

kumquat juice 400ml

100ml of 10% sucrose aqueous solution

Black tea fungus mother liquor 1500ml

Ventilating, standing, fermenting and culturing at 20 ℃, and stopping fermentation when the pH value of a fermentation solution is 3.5-4.5;

5) blending: adding stevioside serving as a 'natural' raw material into the fermentation liquor to replace the traditional cane sugar for seasoning until the sugar degree is 18 degrees Bx; 6) and (3) sterilization: the sterilizing requirement can be met by instantly sterilizing at the high temperature of 95 ℃ for 15s, then carrying out aseptic hot filling, and cooling to 38 ℃ to obtain the cyclocarya paliurus kumquat black tea fungus probiotic beverage.

Example 4 cyclocarya paliurus kumquat and black tea fungus probiotic beverage preparation

1) Preparing cyclocarya paliurus tea: weighing 40g parts of cyclocarya paliurus tea, soaking in 4000ml of boiling water for 10min to prepare tea water, and filtering to obtain tea water filtrate;

2) preparing a black tea fungus mother solution: adding 300g sucrose into 3000ml tea water filtrate obtained in step 1), stirring for dissolving, cooling, adding 150ml black tea fungus starter (acetic acid bacteria 4.0 × 10)⁶cfu/mL; lactic acid bacteria 4.0X 10⁶cfu/mL; yeast 2.0 × 10⁶cfu/mL), standing and ventilating at 25 ℃ until a film appears on the surface, namely the black tea fungus mother liquor, for later use;

3) preparation of kumquat raw juice

The kumquat raw juice prepared in example 1 was used directly.

4) Fermentation: firstly, putting sucrose into a clean glass ware, pouring a proper amount of boiling water, stirring and dissolving, and then transferring to a volumetric flask to fix the volume to prepare a 10% sucrose syrup aqueous solution; adding kumquat juice, cooling, and inoculating black tea fungus mother liquor, wherein the volume of each component is as follows:

300ml of kumquat raw juice

100ml of 10% sucrose aqueous solution

1600ml of black tea fungus mother liquor

Ventilating, standing, fermenting and culturing at 20 ℃, and stopping fermentation when the pH value of a fermentation solution is 3.5-4.5;

5) blending: stevioside, a "natural" material, was added to the fermentation broth to replace traditional sucrose to flavor to a sugar level of 18 Bx. Most of sugar in the fermented kumquat and black tea fungus composite fermentation liquid is consumed by microorganisms, and natural sweet substances are selected to prepare the beverage so as to improve the flavor of the beverage.

6) And (3) sterilization: the sterilizing requirement can be met by instantly sterilizing at the high temperature of 95 ℃ for 15s, then carrying out aseptic hot filling, and cooling to 38 ℃ to obtain the cyclocarya paliurus kumquat black tea fungus probiotic beverage.

Example 5 preparation of control Black tea soup

1) Preparing black tea water: weighing 40g of black tea, soaking in 4000ml of boiling water for 10min to prepare tea water, and filtering to obtain tea water filtrate; 2) preparing a black tea fungus mother solution: adding 300g sucrose into 3000ml tea water filtrate obtained in step 1), stirring for dissolving, cooling, adding 150ml black tea fungus starter (acetic acid bacteria 4.0 × 10)⁶cfu/mL; lactic acid bacteria 4.0X 10⁶cfu/mL; yeast 2.0 × 10⁶cfu/mL), standing and ventilating at 25 ℃ until a film appears on the surface, namely the black tea fungus mother liquor, for later use;

3) preparation of kumquat raw juice

The kumquat raw juice prepared in example 1 was used directly.

4) Fermentation: firstly, putting sucrose into a clean glass ware, pouring a proper amount of boiling water, stirring and dissolving, and then transferring to a volumetric flask to fix the volume to prepare a 10% sucrose syrup aqueous solution; adding kumquat juice, cooling, and inoculating black tea fungus mother liquor, wherein the added content of each component is as follows:

300ml of kumquat raw juice

100ml of 10% sucrose aqueous solution

1600ml of black tea fungus mother liquor

Ventilating, standing, fermenting and culturing at 20 ℃, and stopping fermentation when the pH value of a fermentation solution is 3.5-4.5;

5) blending: adding stevioside serving as a 'natural' raw material into the fermentation liquor to replace the traditional cane sugar for seasoning until the sugar degree is 18 degrees Bx;

6) and (3) sterilization: the sterilization requirement can be met by instantly sterilizing at 90 ℃ for 15s, then carrying out sterile hot filling, and cooling to 38 ℃ to obtain the black tea soup fermented beverage.

Oxidation resistance test

Determination of DPPH radical scavenging Rate:

1) preparing a 0.2mmol/L ethanol DPPH solution: 19.7mg of DPPH is weighed and dissolved by absolute ethyl alcohol, the volume is determined to be 250ml, and the brown bottle is stored in dark place.

2) Sample DPPH free radical scavenging assay reagents added to each group: each group was run in triplicate.

(ii) sample set (A)₁): 2ml of the sample was placed in a test tube with a plug, and 2ml of a 0.2mmol/L DPPH ethanol solution was added.

② sample blank group (A)₂): 2ml of sample is put into a test tube with a plug scale, and then 2ml of absolute ethyl alcohol is added.

③ control group (A)₀): 2ml of absolute ethanol is put into a test tube with a plug scale, and then 2ml of 0.2mmol/L DPPH ethanol solution is added.

3) Mixing uniformly, reacting in dark room at room temperature for 30min, measuring absorbance at 517nm, and calculating DPPH free radical clearance (%). 2. Superoxide anion (O)^2-) Determination of clearance rate:

1) preparing 0.05mol/L Tris-HCl (pH8.2) buffer solution: 3.028g Tris is weighed, added with about 450ml pure water to dissolve, added with a proper amount of concentrated HCl, and the pH is measured by a pH meter while adding, and then the volume is adjusted to 500ml by pure water.

Preparing a pyrogallol solution with the concentration of 6 mmol/L: 75.7mg of pyrogallol reagent is weighed and dissolved by 10mmol/L of dilute hydrochloric acid, and the volume is determined to be 100 ml.

2) Sample superoxide anion scavenging experiment each set of reagents added: each group was run in triplicate.

(ii) sample set (A)₁): 4.5ml of 0.05mol/L Tris-HCl (pH8.2) buffer solution is put into a test tube with a plug scale, water bath is carried out at 25 ℃ for 10min, and then 1ml of sample and 0.4ml of 6mmol/L pyrogallol solution are added.

② sample blank group (A)₂): 4.5ml of 0.05mol/L Tris-HCl (pH8.2) buffer solution is put into a test tube with a plug scale, water bath is carried out at 25 ℃ for 10min, and then 1ml of sample and 0.4ml of pure water are added.

③ control group (A)₀): 4.5ml of 0.05mol/L Tris-HCl (pH8.2) buffer solution is put into a test tube with a plug scale, water bath is carried out at 25 ℃ for 10min, and then 1ml of pure water and 0.4ml of 6mmol/L pyrogallol solution are added.

3) Mixing uniformly, standing at normal temperature for reaction for 4min, adding 1ml of 8mol/L HCl solution respectively to stop the reaction, measuring absorbance at 320nm, and calculating superoxide anion clearance (%).

3. Hydroxyl radical (OH)^-) Determination of clearance rate:

1) preparing 9mmol/L FeSO₄Solution: 0.25g of FeSO was weighed₄Dissolving with pure water, and diluting to 100 ml.

Preparing 9mmol/L salicylic acid ethanol solution: 0.124g of salicylic acid reagent is weighed, dissolved by absolute ethyl alcohol and is added to 100 ml.

③ preparation of 8.8mmol/L H₂O₂Solution: aspirate 90. mu.l of 30% H₂O₂The solution was made to 100ml with pure water.

2) Sample hydroxyl radical scavenging experiment reagents added in each group: each group was run in triplicate.

(ii) sample set (A)₁): 1ml of sample, 1ml of 9mmol/LFeSO₄Solution, 1ml of 9mmol/L ethanol salicylateThe solution was placed in a test tube with a stoppered scale.

② sample blank group (A)₂): 1ml of sample, 1ml of 9mmol/LFeSO₄The solution and 1ml of pure water were put into a test tube with a stopper.

③ control group (A)₀): 1ml of pure water and 1ml of 9mmol/LFeSO were taken₄The solution, 1ml of 9mmol/L ethanol solution of salicylic acid, was placed in a test tube with a stoppered scale.

3) Mixing, adding 1ml of 8.8mmol/L H₂O₂The solution starts to react, water bath is carried out at 37 ℃ for 30min, absorbance is measured at 512nm, and hydroxyl radical clearance (%) is calculated.

Examples 1, 2, 3 after 7 days of fermentation were tested for DPPH clearance of 84%, 78%, 81%, respectively; superoxide anion (O)^2-) The removal capacity is 38%, 32% and 36%; hydroxyl radical (OH)^-) Clearing capacity 51%, 42%, 48%. Comparative fermentation 7d fermented black tea fungus tea soup with black tea as fermentation raw material, DPPH scavenging ability 52%, superoxide anion (O)^2-) Scavenging capacity of 23%, hydroxyl radical (OH)^-) The clearance capacity is increased by 33%. The product obtained by fermenting the black tea fungus by using the cyclocarya paliurus tea has stronger antioxidant activity, and the test result is shown in the following table 1.

TABLE 1

Comprehensive sensory evaluation test

The Qingqian willow kumquat and black tea fungus composite fermented drink prepared in examples 2 to 4 was scored by 10 professionals according to the sensory evaluation criteria of Table 2, the maximum value and the minimum value were discarded, and the average value was calculated, and the results are shown in Table 3:

TABLE 2

TABLE 3

	Color/minute	Flavor/score	Taste/score	Tissue status/score	Total score/minute
						Example 2	18	17	25	28	88
Example 3	17	19	26.5	27.5	90
						Example 4	16.5	18	27	27.5	89

As can be seen from Table 3, the cyclocarya paliurus and kumquat black tea fungus composite fermented beverage prepared under the conditions has the advantages of harmonious taste, moderate sweet and sour taste, unique fragrance, brownish yellow color, clearness, transparency, uniformity and stability, and can emit the fermented fragrance.