Enzymolysis low-temperature extraction bear gall body-building wine and preparation method thereof

Abstract

The invention relates to the technical field of health food processing, and discloses an enzymolysis low-temperature extraction bear gall body-building wine and a preparation method thereof.The preparation method comprises the steps that fructus lycii and fresh bear bile are mixed in a water phase, and bile endogenous cholate is utilized for self-assembly to form micelles; carrying out first-stage enzymolysis-ultrasonic synergistic extraction under low ionic strength, and preferentially extracting weak-polarity components; then adding an ionic strength regulator to improve the ionic strength of the system, and inducing the micelle structure to reconstruct; and carrying out second-stage extraction in the reconstructed high-ion-strength system, and extracting strong-polarity components in a targeted manner. And carrying out enzyme inactivation, and alcoholizing and aging all the materials to obtain the finished wine. According to the method disclosed by the invention, by constructing an ionic strength response dynamic extraction system, efficient and targeted extraction of the functional components is realized under a low-temperature condition, degradation of thermosensitive substances is avoided, and the finally obtained finished wine is high in functional component content, clear and stable in wine body and harmonious in taste.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of health food processing, in particular to an enzyme hydrolysis low-temperature extraction bear bile body-building wine and a preparation method thereof. BACKGROUND

[0002] Bear bile and medlar are both valuable resources of traditional medicine and food, and preparing them into health wine is a common way to exert their health value. At present, the preparation method of related products mainly adopts the traditional long-time soaking method, that is, the raw materials are soaked in high-concentration liquor for several months or even longer. However, this method has significant disadvantages: the extraction process completely depends on the passive penetration of the solvent, resulting in a long extraction period and low efficiency, and a large amount of active ingredients such as medlar polysaccharide and betaine remain in the residue, causing great waste of raw materials.

[0003] To improve the extraction efficiency, some processes introduce heating means such as heat reflux or decoction. Although heating can accelerate molecular motion and shorten the extraction time to a certain extent, the high-temperature environment will inevitably damage the heat-sensitive active substances in medlar and bear bile, and also introduce the adverse flavor caused by boiling, affecting the quality of the final product. In addition, the existing extraction technology usually adopts a single solvent system, which is difficult to simultaneously and efficiently extract a variety of active ingredients with large differences in polarity in medlar, such as weakly polar carotenoids and strongly polar polysaccharides. This non-selective and inefficient extraction method not only causes the loss of active ingredients, but also often leads to turbidity and precipitation of the obtained wine during storage due to the precipitation of substances, poor product stability, and poor integration of the bitter flavor of bear bile and the wine body, making the taste difficult to coordinate. Therefore, the present application provides an enzyme hydrolysis low-temperature extraction bear bile body-building wine and a preparation method thereof to solve the problems in the prior art. SUMMARY

[0004] In view of the deficiencies in the prior art, the present application provides an enzyme hydrolysis low-temperature extraction bear bile body-building wine and a preparation method thereof, which solves the problems in the background art.

[0005] To achieve the above-mentioned purposes, the present application is implemented by the following technical solutions: an enzyme hydrolysis low-temperature extraction bear bile body-building wine, which is prepared from the following raw materials in the following proportions by weight: medlar 50-100 parts; fresh bear bile 1-5 parts; honey 20-40 parts; and corn and sorghum wine 500-600 parts. The fresh bear bile, medlar, honey, and corn and sorghum wine are combined in a specific ratio to provide a material basis for subsequent efficient and targeted extraction under low-temperature conditions, and the final product has high active ingredient content, clear and stable wine body, and coordinated flavor.

[0006] A preparation method of enzymatic low-temperature extraction of bear gall health wine, applied to the above-mentioned enzymatic low-temperature extraction of bear gall health wine, comprising the following steps: (1) Mix wolfberry and fresh bear bile in purified water to construct a water phase extraction system containing bile salt micelles; (2) Perform first-stage enzymatic-ultrasonic synergistic extraction in the water phase extraction system; (3) Add an ion strength regulator to the system after the first-stage extraction is completed to increase the ion strength of the system; (4) Perform second-stage enzymatic-ultrasonic synergistic extraction in the system with increased ion strength to obtain an extract material containing wolfberry residues; (5) Mix the extract material with sorghum, corn wine, and honey to perform alcoholization aging; In step (1), fresh bear bile is placed in the water phase together with wolfberry powder at the initial stage of the extraction process. The bile acid salt molecules in the fresh bear bile have a concentration higher than the critical micelle concentration, and form bile salt micelles in situ in the system. The micelles form a large number of hydrophobic core solubilization microzones in the aqueous solution.

[0007] In step (2), under the initial environment of low ion strength, the formed bile salt micelle structure is relatively loose, and its hydrophobic core preferentially captures and solubilizes weakly polar or nonpolar components in the wolfberry matrix. At the same time, the enzymatic preparation breaks the cell wall of the plant, and the cavitation effect of ultrasonic waves promotes the mass transfer process of these components from the solid matrix to the micelle core.

[0008] In steps (3) and (4), an ion strength regulator is introduced into the system. The presence of a large number of electrolyte ions compresses the electric double layer between the hydrophilic head groups of the micelle surface, producing a salting-out effect, which causes the micelles to restructure into smaller secondary micelles with a more compact structure. The change in the microenvironment is conducive to the extraction and dispersion of components with stronger polarity or amphiphilic properties in wolfberry. Through two stages of extraction, step-by-step, targeted, and efficient extraction of different polar target components in the same reaction kettle is achieved.

[0009] In step (5), full-material alcoholization aging is used, which avoids the loss of functional components adsorbed on the medicinal residues due to solid-liquid separation in traditional processes. High-concentration alcohol can destroy the micelle structure, allowing various components to be fully released and dissolved in the wine body, ensuring the maximum utilization of functional components and also giving the finished wine a full flavor.

[0010] Preferably, in step (1), the wolfberry is superfine pulverized and sieved through 80-120 mesh screen before mixing with fresh bear bile. Superfine pulverization of the wolfberry significantly increases the specific surface area of the material, increasing its contact area with the extraction medium and enzyme preparation, thereby improving the mass transfer efficiency and reaction rate of the subsequent extraction process.

[0011] Preferably, the preparation method has a material-to-liquid ratio of 1:15 to 1:25 (w / v) between the wolfberry and purified water in step (1), which ensures that the material forms a slurry with suitable fluidity in the reaction kettle, facilitating heat and mass transfer, while ensuring that the concentrations of various components in the extraction system are within the high-efficiency reaction range.

[0012] Preferably, the process parameters for the first-stage extraction in step (2) are: temperature 38-45℃, pH 4.5-5.5, ultrasonic frequency 25-40kHz, acoustic power density 100-150W / L, time 40-60 minutes. The low temperature of 38-45℃ can ensure the activity of the enzyme preparation and maintain the stability of the bile salt micelle structure; the pH range of 4.5-5.5 is the optimal environment for the action of the complex enzyme system; the introduction of ultrasonic waves intensifies the extraction process through its mechanical and cavitation effects.

[0013] Preferably, the ionic strength adjusting agent in step (3) is an aqueous solution of sodium chloride or potassium chloride, which makes the final ionic strength of the system reach 0.1-0.2mol / L. Sodium chloride or potassium chloride is a commonly used food industry regulator with high safety, and an ionic strength range of 0.1-0.2mol / L induces effective reconfiguration of the bile salt micelles without causing excessive salting-out precipitation of macromolecules such as proteins in the system.

[0014] Preferably, the process parameters for the second-stage extraction in step (4) are: temperature 38-45℃, time 40-60 minutes. The extraction is carried out at the same low temperature as the first stage to avoid damage to heat-sensitive functional ingredients at high temperatures, ensuring the quality of the final product.

[0015] Preferably, the enzymolysis in step (2) is achieved by adding a complex enzyme preparation, which contains cellulase and pectinase, with a total addition amount of 0.5-1.5% of the dry weight of the wolfberry. The synergistic action of cellulase and pectinase can efficiently hydrolyze the main components of the cell wall and intercellular substance of the wolfberry, breaking down the physical barrier and allowing the intracellular contents to be fully released.

[0016] Preferably, in step (4), the obtained extract is heated to 85-95 DEG C and maintained for 10-15 minutes to deactivate the enzyme, the instant high temperature treatment can completely deactivate the residual enzyme preparation in the system, terminate the enzymatic reaction, avoid the problems of turbidity of wine body and generation of peculiar smell due to continuous action of the enzyme in subsequent aging process, and ensure long-term stability of the product.

[0017] Preferably, the process parameters of the alcohol aging in step (5) are as follows: aging temperature 15-25 DEG C, and aging time 90-180 days. Aging at suitable temperature and for sufficient time is helpful to esterification, condensation and other series of complex physical and chemical changes of water, alcohol and various flavor substances from the raw materials in the wine body, so that the wine body becomes mellow and smooth, and the aroma is more coordinated.

[0018] The application provides an enzyme hydrolysis low-temperature extraction bear bile health wine and a preparation method thereof. 1. The application adds fresh bear bile at the initial extraction stage, uses endogenous bile salt to construct a bile salt micelle micro-reaction field in situ, and realizes step-by-step targeted extraction of different polar active ingredients through subsequent ion intensity response regulation, so that the total extraction rate of various active ingredients such as gypenosides, betaine and carotenoids is significantly improved.

[0019] 2. The whole extraction process is carried out at a low temperature of 38-45 DEG C in combination with enzyme hydrolysis and ultrasonic wave synergistic effect, the mild process conditions effectively avoid the damage to heat-sensitive active substances in gynostemma pentaphyllum and fresh bear bile in the traditional hot reflux or decoction process, so that the biological activity and natural flavor of the raw materials are maximally retained, and the quality of the final product is improved.

[0020] 3. The application uses the solubilization of endogenous bile salt micelles, and adopts a process of directly alcohol aging of the whole material after extraction, so that various active ingredients are uniformly dispersed in the wine body in a stable state, therefore, the final product is clear and bright, and is not easy to produce precipitation during long-term storage, and the wine body flavor is full and the taste is coordinated, and the problem of too prominent bitterness of traditional bear bile products is effectively improved. BRIEF DESCRIPTION OF DRAWINGS

[0021] Figure 1 The application provides a bear bile health wine preparation method flow chart. DETAILED DESCRIPTION

[0022] The technical solutions in the embodiments of the application will be clearly and completely described below with reference to the drawings in the specification of the application. Obviously, the described embodiments are only part of the embodiments of the application, rather than all the embodiments of the application. Based on the embodiments in the application, all other embodiments obtained by those skilled in the art without creative labor fall within the protection scope of the application.

[0023] Reference to the accompanying drawings Figure 1 The embodiment of the present application provides an enzymatic low-temperature extraction bear gall health wine, and the main raw materials and reagent sources and specifications used in the following examples and comparative examples are as follows, and the reagents not specifically described are commercially available analytical pure or higher grade products: Medlar: 50-100 parts, the medlar used is the dried mature fruit of Lycium barbarum L. of Solanaceae, and the market is superior grade product; Fresh bear bile: 1-5 parts, the fresh bear bile used is the liquid obtained by dissolving and recovering the dried bile of Ursidae animal black bear provided by the artificial breeding base in accordance with the relevant national regulations, or the liquid bile directly obtained; Honey: 20-40 parts, the honey used is the natural sweet substance obtained by collecting the nectar, secretion or honeydew of plants by Apidae insects Chinese honeybee or Italian bee, mixing with its own secretion, and fully brewing; Sorghum and corn wine: 500-600 parts; Tauroursodeoxycholic acid (CAS No. 14605-22-2); The present application preferably uses commercially available first-grade acacia honey, and the sorghum and corn wine used is pure grain base wine made of sorghum and corn as raw materials through solid-state fermentation, distillation, aging and blending, with an alcohol content of 55-65% (v / v), which meets the relevant regulations of GB / T10781.1 for superior quality clear type liquor; The cellulase (CAS No. 9012-54-8, EC 3.2.1.4) used is a commercially available food-grade enzyme preparation, with an enzyme activity potency range of 50,000-80,000 U / g; The pectinase (CAS No. 9032-75-1, EC 3.2.1.15) used is a commercially available food-grade enzyme preparation, with an enzyme activity potency range of 30,000-50,000 U / g; The sodium chloride (CAS No. 7647-14-5) used is an analytical pure product commercially available, with a chemical formula of NaCl; The citric acid (CAS No. 77-92-9) used is a commercially available food-grade anhydrous citric acid; The sodium citrate used is trisodium citrate dihydrate (CAS No. 6132-04-3), which is a commercially available food-grade product.

[0024] Example 1 The embodiment provides a preparation method of bear gall health wine based on ion intensity response regulation, and the raw materials used are as follows in terms of weight fraction: medlar 50 parts, fresh bear bile 1 part, honey 20 parts, and sorghum and corn wine with an alcohol content of 60% (v / v) 500 parts. The method specifically comprises the following steps: (1) 50 parts of wolfberry fruit are ultra-micronized and passed through a 120-mesh screen to obtain wolfberry fruit powder.

[0025] (2) 750 parts of purified water are added to a reaction kettle, stirring is started and the temperature is raised to 38°C, the wolfberry fruit powder of step (1) and 1 part of fresh bear gall are added, stirring is uniform, and the pH value of the system is adjusted to 5.5 using citric acid.

[0026] (3) A composite enzyme preparation accounting for 0.5% of the dry weight of wolfberry fruit is added to the system, the cellulase and pectinase are mixed at a mass ratio of 2:1, the ultrasonic wave is turned on, the frequency is 25 kHz, the acoustic power density is 100 W / L, the first stage extraction is carried out at 38°C, and the duration is 40 minutes.

[0027] (4) The temperature, stirring and ultrasonic state are kept unchanged, and 1.0 mol / L of sodium chloride solution is quickly pumped into the reaction kettle to make the final concentration of sodium chloride in the system reach 0.1 mol / L.

[0028] (5) The second stage extraction is continued under the conditions of step (4) for 40 minutes.

[0029] (6) After the extraction is completed, the material in the kettle is raised to 85°C and maintained for 15 minutes to completely inactivate the enzyme, and then cooled to room temperature.

[0030] (7) All the materials in the kettle are pumped into a aging tank, 500 parts of sorghum and corn wine with an alcohol content of 60% (v / v) and 20 parts of honey are added, stirring is uniform, and then sealed.

[0031] (8) The aging is carried out at 20°C in the dark for 90 days, after the aging is completed, the residue is removed by plate and frame filter to obtain the clear finished product wine.

[0032] Example 2: The present embodiment provides a preparation method of bear gall health wine based on ion strength response regulation, and the raw materials used are 100 parts of wolfberry fruit, 5 parts of fresh bear gall, 40 parts of honey, and 600 parts of sorghum and corn wine with an alcohol content of 65% (v / v), and the method comprises the following steps: (1) 100 parts of wolfberry fruit are ultra-micronized and passed through an 80-mesh screen to obtain wolfberry fruit powder; (2) 2500 parts of purified water are added to a reaction kettle, stirring is started and the temperature is raised to 45°C, the wolfberry fruit powder of step (1) and 5 parts of fresh bear gall are added, stirring is uniform, and the pH value of the system is adjusted to 4.5 using citric acid; (3) A composite enzyme preparation accounting for 1.5% of the dry weight of wolfberry fruit is added to the system, the cellulase and pectinase are mixed at a mass ratio of 2:1, the ultrasonic wave is turned on, the frequency is 40 kHz, the acoustic power density is 150 W / L, the first stage extraction is carried out at 45°C, and the duration is 60 minutes; (4) keeping the temperature, stirring and ultrasonic state unchanged, quickly pump 1.2 mol / L sodium chloride solution into the reactor, so that the final concentration of sodium chloride in the system reaches 0.2 mol / L; (5) continue the second stage extraction under the condition of step (4) for 60 minutes; (6) after the extraction is completed, the material in the reactor is heated to 95℃ and maintained for 10 minutes to completely inactivate the enzyme, and then cooled to room temperature; (7) pump all the materials in the reactor into the aging tank, add 600 parts of sorghum, corn wine with an alcohol content of 65% (v / v) and 40 parts of honey, stir uniformly and seal; (8) store in a dark environment at 15℃ for 180 days, and then filter the residue through a diatomite filter to obtain a clear finished product wine.

[0033] Example 3: The present embodiment provides a preparation method of bear gall fitness wine based on ion strength response regulation, and the raw materials used are as follows in terms of weight fraction: 75 parts of wolfberry, 3 parts of fresh bear bile, 30 parts of honey, 550 parts of sorghum and corn wine with an alcohol content of 60% (v / v), which comprises the following steps: (1) ultra-micro powder 75 parts of wolfberry, pass through a 100 mesh screen to obtain wolfberry powder; (2) add 1500 parts of purified water to the reactor, start stirring and heat to 42℃, add the wolfberry powder of step (1) and 3 parts of fresh bear bile, stir uniformly, and use citric acid to adjust the pH value of the system to 5.0; (3) add 1.0% of a composite enzyme preparation based on the dry weight of wolfberry to the system, mix cellulase and pectinase at a mass ratio of 2:1, start ultrasonic wave with a frequency of 30 kHz and a sound power density of 125 W / L, and carry out the first stage extraction at 42℃ for 50 minutes; (4) keep the temperature, stirring and ultrasonic state unchanged, quickly pump 1.0 mol / L potassium chloride solution into the reactor, so that the final concentration of potassium chloride in the system reaches 0.15 mol / L; (5) continue the second stage extraction under the condition of step (4) for 50 minutes; (6) after the extraction is completed, the material in the reactor is heated to 90℃ and maintained for 12 minutes to completely inactivate the enzyme, and then cooled to room temperature; (7) pump all the materials in the reactor into the aging tank, add 550 parts of sorghum and corn wine with an alcohol content of 60% (v / v) and 30 parts of honey, stir uniformly and seal; (8) store in a dark environment at 22℃ for 120 days, and then filter the residue through a plate and frame filter to obtain a clear finished product wine.

[0034] Comparative Example 1: Compared with Example 1, the difference lies in that a traditional soaking method is adopted: 50 parts of wolfberry fruit, which is crushed by the same method, is directly added into 500 parts of sorghum and corn wine, and then the mixture is soaked in a dark environment at 20℃ for 90 days after being sealed; after the soaking is completed, the wine liquid is filtered, and then 1 part of fresh bear bile and 20 parts of honey are added into the wine liquid and stirred uniformly, thereby obtaining the product of Comparative Example 1.

[0035] Comparative Example 2: Compared with Example 1, the difference lies in that the fresh bear bile is not used as a process aid in advance: the fresh bear bile is not added in step (2); after step (7) is completed, the solid-liquid separation is performed on the materials in the kettle, and the supernatant is obtained; in step (7), the supernatant is mixed with 500 parts of sorghum and corn wine, 20 parts of honey and 1 part of fresh bear bile, and the remaining steps and parameters are the same.

[0036] Comparative Example 3: Compared with Example 1, the difference lies in that the ion strength response regulation is not performed: the operation of adding the sodium chloride solution in step (4) is omitted; in order to ensure that the total extraction time is consistent, the first-stage extraction time in step (3) is extended to 80 minutes, and the remaining steps and parameters are the same.

[0037] Comparative Example 4: Compared with Comparative Example 2, the difference lies in that an exogenous surfactant is used to perform the solubilization extraction: in step (2) of Comparative Example 2, Tween-80 is added into the system at an equivalent molar concentration of the bile acid salt in the fresh bear bile in Example 1, and the remaining steps and parameters are the same as those in Comparative Example 2.

[0038] Test Example 1: The test example aims to verify that the present application can realize the time-sharing and targeted extraction of different polar target components by regulating the ion strength, and the experimental object is the preparation process of Example 2. By analyzing the samples at two key time points in the extraction process, the dissolution rules of weakly polar and strongly polar components are characterized.

[0039] Experimental steps: Sample preparation: according to the method described in Example 2, a part of the liquid material is taken out from the reaction kettle before the first-stage extraction is completed and the sodium chloride solution is pumped in, and the supernatant is taken after centrifugation at 10000 r / min for 10 minutes, which is marked as sample A. A sample is taken from the reaction kettle again before the second-stage extraction is completed and the enzyme inactivation treatment is performed, and the supernatant is taken after centrifugation under the same conditions, which is marked as sample B.

[0040] Total carotenoids (weak polar component indicator) content determination: accurately pipette a certain volume of sample A and sample B, add equal volume of n-hexane for liquid-liquid extraction, repeat extraction three times, combine n-hexane phase, after constant volume, measure its absorbance at 450 nm wavelength. Calculate the content of total carotenoids in the sample according to the standard sample curve, unit: mg / L.

[0041] Gouqi polysaccharide (strong polar component indicator) content determination: using phenol-sulfuric acid method, accurately pipette the appropriately diluted sample A and sample B, add phenol solution and concentrated sulfuric acid in turn, after color reaction, measure its absorbance at 490 nm wavelength, calculate the content of gouqi polysaccharide in the sample according to the standard sample curve, unit: mg / L.

[0042] Experimental data: Table 1. Determination results of target component content in sample at different extraction stages The above test data shows that from the end of the first stage (sample A) to the end of the second stage (sample B), the content increment of total carotenoids in the system is 3.33 mg / L, while the content increment of gouqi polysaccharide is as high as 372.3 mg / L.

[0043] The results are consistent with the technical mechanism of the present application. In the low ionic strength environment of the first stage, the micellar structure formed in situ from fresh bear bile is relatively loose, and the unfolded hydrophobic core provides an excellent solubilization microzone for weakly polar carotenoids, which are preferentially dissolved from the gouqi matrix. When the ionic strength of the system increases before the start of the second stage, the salt-out effect causes the electrostatic repulsion between the hydrophilic heads of the micelles to be compressed, and the micellar structure is temporarily restructured into a more compact form, so that the microreaction field environment is conducive to the dissolution and dispersion of the strongly polar gouqi polysaccharide macromolecules.

[0044] Therefore, the results of the test example confirm that the method of regulating the micellar structure by ionic strength response can realize the step-by-step and targeted extraction of different polar target components in a continuous process, proving the technical feasibility of the present application.

[0045] Test example 2: The purpose of this test example is to comprehensively evaluate the superiority of the method of the present application relative to different comparative methods by determining the content of key functional components and physical stability indicators in the finished wine.

[0046] Experimental steps: Sample preparation: prepare the finished wine sample according to the method described in example 1 and comparative example 1, comparative example 2, comparative example 3, comparative example 4, respectively.

[0047] Functional component content determination: Tauroursodeoxycholic acid (TUDCA) content determination: HPLC was used, and each sample was filtered through a 0.22 μm microporous filter and then injected. The chromatographic column was a C18 column (4.6 mm x 250 mm, 5 μm); the mobile phase was methanol-0.02 mol / L potassium dihydrogen phosphate solution (70:30, v / v); the flow rate was 1.0 mL / min; the detection wavelength was 210 nm; and the column temperature was 30°C. The TUDCA content was calculated according to the external standard method.

[0048] Total carotenoid content determination: Each sample was accurately pipetted and liquid-liquid extracted with n-hexane. The n-hexane phase was combined and diluted to volume, and the absorbance was determined at 450 nm. The content was calculated according to the standard curve.

[0049] Lycium barbarum polysaccharide content determination: The phenol-sulfuric acid method was used. Each sample was accurately pipetted after appropriate dilution, and the absorbance was determined at 490 nm after color development. The content was calculated according to the standard curve.

[0050] Physical stability determination: Each finished wine sample was sealed and placed in a 4°C constant temperature environment for 7 days. After being taken out and restored to room temperature, the turbidity value (NTU) was determined using a turbidimeter, so as to evaluate the low-temperature stability.

[0051] Experimental data: Table 2. Detection results of key indicators of finished wine of each example and comparative example According to the data in Table 2, the content of each functional ingredient of the finished wine prepared by Example 1 of the present application is significantly higher than that of all comparative examples, and the turbidity after low-temperature standing is the lowest, indicating that the product quality and stability are overall superior.

[0052] Compared with Comparative Example 1 (traditional soaking method), the content of each functional ingredient of Example 1 is increased by several times, proving that the enzyme-ultrasound synergistic extraction process of the present application is much better than the traditional passive soaking.

[0053] Compared with Comparative Example 2 (post-addition of bear bile), the contents of carotenoids and lycium barbarum polysaccharide of Example 1 are increased by about 68% and 33%, respectively, and the turbidity is significantly reduced, proving that the prepositioning of fresh bear bile as a process additive to construct a micellar micro-reaction field is the key to improving the extraction efficiency and product stability, rather than simply adding ingredients.

[0054] The comparison with Comparative Example 3 (without ion intensity response regulation) is particularly key. Under the same total extraction time, the content of lycium barbarum polysaccharide of Example 1 is increased by about 95% compared with Comparative Example 3. This result strongly confirms the core mechanism of the present application: by regulating the micellar structure through ion intensity improvement, targeted and efficient extraction of strong polar polysaccharide ingredients can be achieved, and the lack of this step will lead to incomplete extraction of functional ingredients.

[0055] Compared with Comparative Example 4 (using an exogenous surfactant), Example 1 not only contains the efficacy component TUDCA of bear bile itself, but also has better comprehensive extraction efficiency of endogenous components of wolfberry and stability of the final product, indicating that the self-assembly and response characteristics of the endogenous bile salt system of fresh bear bile are better than those of conventional non-specific solubilizers.

[0056] Test Example 4: The present test example aims to evaluate the comprehensive quality of the product of the present application relative to the product of the traditional process in terms of color, aroma, taste and clarity, etc. through professional sensory evaluation.

[0057] Experimental steps: Sample preparation: Take the finished products prepared in Example 1, Comparative Example 1 and Comparative Example 2 as the samples to be evaluated, and place them in a 20°C constant temperature environment after being uniformly numbered.

[0058] Evaluation team: Select 10 personnel who have been trained in professional sensory evaluation and are familiar with wine evaluation to form the evaluation team.

[0059] Evaluation method: Use double-blind method, pour the samples into standard wine glasses, and have the evaluation team refer to the following scoring standards to score the color, aroma, taste and clarity of each sample independently (full score is 10).

[0060] Color: uniform color, clear and transparent amber or golden yellow is the best.

[0061] Aroma: unique aroma of bear bile, harmonious integration of fruit aroma and wine aroma of wolfberry, no foreign odor.

[0062] Taste: smooth and smooth taste, aftertaste, moderate bitterness of bear bile without prominence.

[0063] Clarity: clear and transparent wine body, no suspended matter, no precipitate.

[0064] Data processing: collect all scores, remove the highest and lowest scores, and calculate the average score of each index.

[0065] Experimental data: Table 3. Average scores of sensory evaluation of different samples The sensory evaluation results show that the scores of Example 1 sample in all evaluation dimensions are significantly higher than those of Comparative Example 1 and Comparative Example 2.

[0066] Compared with Comparative Example 1 (traditional soaking method), the advantages of Example 1 in aroma and taste are particularly prominent. This is due to the low-temperature extraction and full-material aging process of the present application, which not only efficiently extracts flavor substances, but more importantly, the encapsulation of bitter components by bile salt micelles and their slow release and transformation during the aging process, so that the bitterness of bear bile is well integrated with the wine body, no longer prominent, achieving the harmony and coordination of the taste.

[0067] Compared with Comparative Example 2 (postpositioning of bear bile), Example 1 performs better in clarity and taste. This shows that the micellar system formed by the fresh bear bile as a functional additive plays a key role in the stable dispersion of various components, which ensures the long-term clarity of the wine body. Simply adding bear bile as a component in the later stage cannot play this stabilizing role, which can easily lead to the precipitation of substances and the stratification of taste.

Claims

1. An enzyme hydrolysis low-temperature extraction bear gall health wine, characterized in that, The following raw materials are included by weight: Wolfberry: 50-100 parts; Fresh bear bile: 1-5 parts; Honey: 20-40 parts; Sorghum and corn wine: 500-600 parts.

2. A preparation method of enzymatic low-temperature extraction of bear gall health wine, applied to the bear gall health wine of claim 1, characterized in that, The following steps are included: (1) Wolfberry is mixed with fresh bear bile in purified water to create a water phase extraction system containing bile salt micelles; (2) First-stage enzyme-ultrasound synergistic extraction is performed in the water phase extraction system; (3) An ion strength regulator is added to the system after the first-stage extraction to increase the ion strength of the system; (4) Second-stage enzyme-ultrasound synergistic extraction is performed in the system with increased ion strength to obtain an extract material containing wolfberry residue; (5) The extract material is mixed with sorghum and corn wine and honey for alcoholification aging.

3. The preparation method of the enzyme hydrolysis low-temperature extraction bear gall health wine according to claim 2, characterized in that, In step (1), the wolfberry is ultra-finely pulverized and sieved through an 80-120 mesh screen before being mixed with fresh bear bile.

4. The preparation method of the enzyme hydrolysis low-temperature extraction bear gall health wine according to claim 2, characterized in that, In step (1), the ratio of wolfberry to purified water is 1:15 to 1:25 (w / v).

5. The preparation method of the enzyme hydrolysis low-temperature extraction bear gall health wine according to claim 2, characterized in that, In step (2), the process parameters for the first-stage extraction are: temperature 38-45°C, pH 4.5-5.5, ultrasonic frequency 25-40 kHz, acoustic power density 100-150 W / L, and time 40-60 minutes.

6. The preparation method of the enzyme hydrolysis low-temperature extraction bear gall health wine according to claim 2, characterized in that, In step (3), the ion strength regulator is an aqueous solution of sodium chloride or potassium chloride, and the final ion strength of the system after addition is 0.1-0.2 mol / L.

7. The preparation method of the enzyme hydrolysis low-temperature extraction bear gall health wine of claim 2, characterized in that, In step (4), the process parameters for the second-stage extraction are: temperature 38-45°C and time 40-60 minutes.

8. The preparation method of the enzyme hydrolysis low-temperature extraction bear gall health wine according to claim 2, characterized in that, The ceramic composite ultrafiltration membrane is prepared by the following steps: In step (2), the enzyme hydrolysis is achieved by adding a composite enzyme preparation containing cellulase and pectinase, with a total addition amount of 0.5-1.5% of the dry weight of wolfberry.

9. The preparation method of the enzyme hydrolysis low-temperature extraction bear gall health wine according to claim 2, characterized in that, In step (4), the obtained extract material is heated to 85-95°C and maintained for 10-15 minutes to inactivate the enzymes.

10. The preparation method of the enzyme hydrolysis low-temperature extraction bear gall health wine according to claim 2, characterized in that, In step (5), the process parameters for alcoholification aging are: aging temperature 15-25°C and aging time 90-180 days.

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