Compound preparation process of multi-effect synergistic enhanced bear gall body-building wine

Abstract

The invention relates to the technical field of health wine preparation, and discloses a multi-effect synergistic enhanced bear gall body-building wine compound preparation process which comprises the following steps: taking sorghum, corn and fructus lycii as fermentation base materials, and carrying out double-strain synergistic fermentation by adopting saccharomyces cerevisiae and lactobacillus plantarum; adding fresh ursodesoxycholic acid during fermentation, carrying out in-situ enzymatic debinding conversion by using a compound enzyme system generated by a fermentation system, and converting the bound bile acid into free ursodesoxycholic acid; after fermentation, distilling to obtain raw wine; a molecularly imprinted polymer prepared by taking a wolfberry polyphenol key monomer as a template molecule is adopted to carry out targeted anchoring refining on raw wine, selective capture and enrichment of wolfberry polyphenol are carried out, and finally aging is carried out to obtain a finished product. According to the method disclosed by the invention, upstream double-strain fermentation in-situ enzymatic conversion and downstream molecular imprinting targeted refining are systematically integrated, so that the technical contradictions that the conversion rate of combined bile acid is low and a large amount of polyphenol functional components are synchronously lost during purification and impurity removal in a traditional process are solved.

Description

Technical Field

[0001] This invention relates to the field of health wine preparation technology, specifically a compound preparation process for a multi-functional synergistic enhancement type of bear bile health wine. Background Technology

[0002] Health-enhancing wines, as a product integrating traditional dietary concepts with modern brewing techniques, aim to regulate bodily functions by incorporating food and medicinal substances with specific health benefits into the wine base. Among the many ingredients, bear bile and wolfberry have attracted much attention due to their clearly documented efficacy. The core active ingredient of bear bile is ursodeoxycholic acid (UDCA), which has a definite effect on protecting the liver and promoting bile secretion; while wolfberry is rich in various active substances such as wolfberry polyphenols, possessing various physiological activities such as antioxidation. Combining these two ingredients to prepare health-enhancing wines, in order to obtain synergistic health benefits, is a development direction in this field.

[0003] Currently, the preparation process of such health wines mainly relies on the traditional soaking method, which involves directly immersing the raw materials in high-proof liquor for prolonged static extraction. However, existing technologies face significant technical bottlenecks in achieving efficient conversion and synergistic retention of active ingredients. Firstly, in bear bile directly derived from organisms, the core active ingredient, ursodeoxycholic acid, exists primarily in a bound form, conjugated with glycine or taurine. This bound bile acid not only has relatively low bioavailability but also possesses a strong bitter and fishy odor, severely degrading the sensory quality of the final product. Traditional soaking, as a passive physical extraction process, cannot facilitate the conversion of bound bile acids into free forms with higher bioactivity and better flavor, thus limiting the product's efficacy and market acceptance.

[0004] Furthermore, for plant-based raw materials such as goji berries, the soaking method, while extracting polyphenols and other functional components, also dissolves a large number of non-target components, such as macromolecular pigments and off-flavor substances, resulting in an overly dark color, rough taste, and uncoordinated aroma in the wine. To improve the taste, existing technologies sometimes use non-specific adsorbent materials such as activated carbon for post-treatment. Although this treatment can remove some off-flavors to a certain extent, its effect lacks selectivity and simultaneously adsorbs and removes valuable polyphenolic functional components and esters, resulting in a simultaneous loss of efficacy and flavor, leading to a thinner body and ultimately being counterproductive. Therefore, how to effectively transform one core functional component while selectively retaining and enriching another, and ultimately achieving a unity of efficacy and sensory quality, is a technical problem that urgently needs to be solved in this field. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a multi-functional synergistic enhancement process for bear bile health wine, which solves the technical contradictions of low conversion efficiency of bound bile acids leading to limited efficacy in traditional processes and the simultaneous and substantial loss of polyphenolic active ingredients due to non-specific adsorption during purification and impurity removal.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a multi-functional synergistic enhancement process for bear bile health wine, comprising the following steps: Step S1: Co-fermentation of the functional substrate with two strains: Sorghum and corn are mixed in a preset ratio, then crushed and passed through a 20-40 mesh sieve. The crushed grains are mixed with whole goji berries to form a fermentation base. Brewing yeast is added to the fermentation base for primary fermentation. When the primary fermentation reaches a certain stage, Lactobacillus plantarum is added for co-fermentation, ultimately yielding a compound fermented mash.

[0007] In one specific implementation, the dry weight ratio of sorghum, corn, and goji berries is 6:3:1. The amount of brewing yeast added is 0.3-0.5% of the total dry weight of the feed, and the amount of *Lactobacillus plantarum* added is 0.1-0.2% of the total dry weight of the feed; wherein, the total dry weight of the feed is defined as the sum of the dry weights of sorghum, corn, and goji berries used in step S1. The addition of *Lactobacillus plantarum* is carried out when the main fermentation temperature rises to 35-38℃.

[0008] The mechanism of this step is as follows: alcoholic fermentation is carried out using brewer's yeast to provide the base wine for subsequent steps; the introduced Lactobacillus plantarum produces flavor substances such as lactic acid to optimize the microenvironment of the mash, while the various extracellular enzymes it produces provide the enzyme source basis for the conversion of bear bile functional components in the next step.

[0009] Step S2: In-situ enzymatic debinding and transformation of the core active ingredient: Fresh bear bile is added to the compound fermented mash obtained in step S1 at a specific temperature, and maintained under specific temperature and time conditions to carry out in situ enzymatic debinding transformation.

[0010] In one specific implementation, the amount of fresh bear bile added is 0.5-1.5% of the total dry weight of the feed. The in-situ enzymatic debinding conversion is controlled at a temperature of 30-35°C for 48-72 hours.

[0011] The mechanism of this step is as follows: using the various hydrolytic enzymes produced by microorganisms in the compound fermentation mash system, as well as the autolytic enzymes contained in bear bile itself, under suitable temperature and pH conditions, they act on the bound bile acids (such as tauroursodeoxycholic acid) in bear bile, causing them to hydrolyze and break down, converting the bound functional core molecules (such as ursodeoxycholic acid) into free states, thereby increasing the proportion of their usable forms in the final product.

[0012] Step S3: Flavor Coupling and Distillation Honey is added to the compound fermented mash after the transformation in step S2 to couple the flavors. The mash is then distilled, and the distillate is collected in segments, with the middle segment of the original wine being extracted.

[0013] In one specific embodiment, the amount of honey added is 2.0-3.0% of the total dry weight of the feed. In the distillation step, the middle section of the original liquor with an alcohol content (volume fraction) of 60-70% vol is collected.

[0014] Step S4: Molecular Imprinting-Based Targeted Anchoring Refining Process The raw wine obtained in step S3 is passed through a chromatography column filled with a molecularly imprinted polymer. First, the main wine liquid A effluent from the chromatography column is collected; then, the molecularly imprinted polymer in the chromatography column is eluted using a specific eluent, and the flavor essence B rich in wolfberry polyphenols eluted during this process is collected; finally, the flavor essence B is added back to the main wine liquid A to complete the refining and recombination.

[0015] The molecularly imprinted polymer used in this step is prepared as follows: chlorogenic acid is used as the template molecule, 4-vinylpyridine as the functional monomer, ethylene glycol dimethacrylate as the crosslinking agent, and azobisisobutyronitrile as the initiator. A thermally initiated polymerization reaction is carried out under specific conditions to obtain a crude polymer. The molar ratio of the template molecule, functional monomer, and crosslinking agent is 1:4:20. The thermally initiated polymerization reaction is carried out at a constant temperature of 60-70°C for 20-28 hours. After polymerization, the crude polymer is subjected to Soxhlet extraction using a methanol-glacial acetic acid mixture with a volume ratio of 9:1 to remove the template molecule, ultimately obtaining the molecularly imprinted polymer with recognition pores capable of specifically recognizing chlorogenic acid and its structural analogs.

[0016] In one specific embodiment, the eluent in step S4 is an aqueous solution of ethanol with a volume fraction of 70-80% and containing 0.5-1.0% glacial acetic acid by volume.

[0017] The mechanism of this step lies in utilizing the imprinted pores on the surface of the molecularly imprinted polymer, which are highly matched with the template molecule (chlorogenic acid, a representative of wolfberry polyphenols) in terms of spatial structure and chemical sites, to achieve specific adsorption and capture of wolfberry polyphenols in the raw wine. When the main wine liquid A flows through, most of the alcohols, water, esters, and the converted bear bile active ingredients pass through, while the wolfberry polyphenols are anchored within the column. Subsequently, by changing the polarity and pH of the eluent, the interaction between the polyphenols and the imprinted sites is disrupted, eluting them out to obtain a high-purity functional flavor essence B. This process achieves targeted separation and enrichment of specific functional components while removing some off-flavor substances from the raw wine.

[0018] Step S5: Aging: The refined and reconstituted liquor from step S4 is aged for 3-6 months in a constant temperature and light-proof environment at 15-20℃ to finally obtain a multi-functional synergistic enhancement bear bile health wine.

[0019] This invention provides a compound formulation process for a multi-functional, synergistically enhanced bear bile health tonic. It possesses the following beneficial effects: 1. A process combining dual-strain synergistic fermentation and in-situ enzymatic debinding and transformation utilizes various hydrolytic enzymes produced by microbial metabolism to act on conjugated bile acids in fresh bear bile, promoting their hydrolysis into free, functional core molecules. This significantly improves the conversion rate and bioavailability of the core functional components of bear bile, solving the technical problem of the ineffective utilization of functional components in traditional processes.

[0020] 2. By employing a targeted anchoring refining process based on molecular imprinting technology, particularly using molecularly imprinted polymers prepared with chlorogenic acid as a template molecule, selective separation and efficient enrichment of the polyphenolic active ingredients in the raw wine were achieved. This method not only effectively avoids the significant loss of these heat-sensitive and easily oxidized active ingredients in traditional processes, but also achieves the technical effect of removing some off-flavor substances and purifying the wine.

[0021] 3. Through a systematic process design that differentiates the active ingredients from different sources (enzymatic transformation, molecular imprinting enrichment) and then refines and recombines them, it is ensured that the core active ingredients in bear bile and goji berries coexist in the final product in a highly active and pure form. After subsequent aging, the various components are fully coordinated and integrated in the wine, resulting in a bear bile health wine with a mellow flavor, harmonious taste, and effective complementarity and synergy among the multiple active ingredients. Detailed Implementation

[0022] The technical solutions of the present invention will be clearly and completely described below with reference to embodiments, comparative examples, and test examples. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0023] Examples 1-3: Example 1: This embodiment provides a compound formulation process for a multi-functional, synergistically enhanced bear bile health wine. The specific steps are as follows: Step S1: Co-fermentation of two strains of active ingredients Sorghum, corn, and goji berries were weighed according to a dry weight ratio of 6:3:1. The sorghum and corn were mixed, crushed, and sieved through a 20-mesh sieve. This mixture was then thoroughly mixed with whole goji berries to form the fermentation base. The total dry weight of the fermentation base was recorded as the total feed dry weight. 0.3% (by total feed dry weight) of brewing yeast was added to the fermentation base for primary fermentation. When the primary fermentation temperature reached 35℃, 0.1% (by total feed dry weight) of *Lactobacillus plantarum* was added for co-fermentation, yielding a compound fermented mash.

[0024] Step S2: In-situ enzymatic deconjugation and transformation of core active ingredients Add 0.5% of the total dry weight of fresh bear bile to the compound fermented mash obtained in step S1, and carry out in-situ enzymatic debinding conversion under constant temperature of 30°C for 48 hours.

[0025] Step S3: Flavor Coupling and Distillation Add 2.0% (by dry weight of the total feed) of honey to the compound fermented mash after step S2 to couple the flavors. Then distill the mash and collect the middle section of the original liquor with an alcohol content of 60% vol.

[0026] Step S4: Targeted Anchoring Refining Process Based on Molecular Imprinting First, the molecularly imprinted polymer was prepared: chlorogenic acid was used as the template molecule, 4-vinylpyridine as the functional monomer, ethylene glycol dimethacrylate as the crosslinking agent, and azobisisobutyronitrile as the initiator. The polymers were mixed in a molar ratio of 1:4:20 and then subjected to thermally initiated polymerization at a constant temperature of 60°C for 20 hours. After polymerization, the polymer was subjected to Soxhlet extraction with a mixed solution of methanol and glacial acetic acid in a volume ratio of 9:1 to remove the template molecule. After drying, the molecularly imprinted polymer was obtained.

[0027] The prepared molecularly imprinted polymer was packed into a chromatography column. The raw wine obtained in step S3 was passed through the chromatography column, and the eluent main wine liquid A was collected. Subsequently, an ethanol-water solution with a volume fraction of 70% ethanol and a volume fraction of 0.5% glacial acetic acid was used as the eluent to elute the chromatography column, and the functional flavor essence B rich in wolfberry polyphenols was collected. Finally, the functional flavor essence B was added back into the main wine liquid A to complete the refining and recombination.

[0028] Step S5: Aging The refined and reconstituted liquor from step S4 is aged in a constant temperature and light-proof environment at 15°C for 3 months to obtain a multi-functional synergistic enhancement bear bile health liquor.

[0029] Example 2: This embodiment provides a compound formulation process for a multi-functional, synergistically enhanced bear bile health wine. The specific steps are as follows: Step S1: Co-fermentation of two strains of active ingredients Sorghum, corn, and goji berries were weighed according to a dry weight ratio of 6:3:1. The sorghum and corn were mixed, crushed, and sieved through a 40-mesh sieve. This mixture was then thoroughly mixed with whole goji berries to form the fermentation base. The total dry weight of the fermentation base was recorded as the total feed dry weight. 0.5% (by total feed dry weight) of brewing yeast was added to the fermentation base for primary fermentation. When the primary fermentation temperature reached 38℃, 0.2% (by total feed dry weight) of *Lactobacillus plantarum* was added for co-fermentation, yielding a compound fermented mash.

[0030] Step S2: In-situ enzymatic deconjugation and transformation of core active ingredients Add 1.5% of the total dry weight of fresh bear bile to the compound fermented mash obtained in step S1, and carry out in-situ enzymatic debinding conversion under constant temperature of 35℃ for 72 hours.

[0031] Step S3: Flavor Coupling and Distillation Add 3.0% (by dry weight of the total feed) of honey to the compound fermented mash after step S2 to couple the flavors. Then distill the mash and collect the middle section of the original liquor with an alcohol content of 70% vol.

[0032] Step S4: Targeted Anchoring Refining Process Based on Molecular Imprinting First, the molecularly imprinted polymer was prepared: chlorogenic acid was used as the template molecule, 4-vinylpyridine as the functional monomer, ethylene glycol dimethacrylate as the crosslinking agent, and azobisisobutyronitrile as the initiator. The polymers were mixed in a molar ratio of 1:4:20 and then subjected to thermally initiated polymerization at a constant temperature of 70°C for 28 hours. After polymerization, the polymer was subjected to Soxhlet extraction using a mixed solution of methanol and glacial acetic acid in a volume ratio of 9:1 to remove the template molecule. After drying, the molecularly imprinted polymer was obtained.

[0033] The prepared molecularly imprinted polymer was packed into a chromatography column. The raw wine obtained in step S3 was passed through the chromatography column, and the eluent main wine liquid A was collected. Subsequently, an ethanol-water solution with an ethanol volume fraction of 80% and containing 1.0% glacial acetic acid was used as the eluent to elute the chromatography column, and the functional flavor essence B rich in wolfberry polyphenols was collected. Finally, the functional flavor essence B was added back into the main wine liquid A to complete the refining and recombination.

[0034] Step S5: Aging The refined and reconstituted liquor from step S4 is aged in a constant temperature and light-proof environment at 20°C for 6 months to obtain a multi-functional synergistic enhancement bear bile health liquor.

[0035] Example 3: This embodiment provides a compound formulation process for a multi-functional, synergistically enhanced bear bile health wine. The specific steps are as follows: Step S1: Co-fermentation of two strains of active ingredients Sorghum, corn, and goji berries were weighed according to a dry weight ratio of 6:3:1. The sorghum and corn were mixed, crushed, and sieved through a 30-mesh sieve. This mixture was then thoroughly mixed with whole goji berries to form the fermentation base. The total dry weight of the fermentation base was recorded as the total feed dry weight. 0.4% (by total feed dry weight) of brewing yeast was added to the fermentation base, and primary fermentation was initiated. When the primary fermentation temperature reached 36.5℃, 0.15% (by total feed dry weight) of *Lactobacillus plantarum* was added for co-fermentation, yielding a compound fermented mash.

[0036] Step S2: In-situ enzymatic deconjugation and transformation of core active ingredients Add 1.0% of the total dry weight of fresh bear bile to the compound fermented mash obtained in step S1, and carry out in-situ enzymatic debinding conversion under constant temperature conditions of 32.5℃ for 60 hours.

[0037] Step S3: Flavor Coupling and Distillation Add 2.5% honey (by dry weight of the total feed) to the compound fermented mash after step S2 to couple the flavors. Then distill the mash and collect the middle section of the original liquor with an alcohol content of 65% vol.

[0038] Step S4: Targeted Anchoring Refining Process Based on Molecular Imprinting First, the molecularly imprinted polymer was prepared: chlorogenic acid was used as the template molecule, 4-vinylpyridine as the functional monomer, ethylene glycol dimethacrylate as the crosslinking agent, and azobisisobutyronitrile as the initiator. The polymers were mixed in a molar ratio of 1:4:20 and then subjected to thermally initiated polymerization at a constant temperature of 65°C for 24 hours. After polymerization, the polymer was subjected to Soxhlet extraction using a mixed solution of methanol and glacial acetic acid in a volume ratio of 9:1 to remove the template molecule. After drying, the molecularly imprinted polymer was obtained.

[0039] The prepared molecularly imprinted polymer was packed into a chromatography column. The raw wine obtained in step S3 was passed through the chromatography column, and the eluent main wine liquid A was collected. Subsequently, an ethanol-water solution with a volume fraction of 75% ethanol and a volume fraction of 0.75% glacial acetic acid was used as the eluent to elute the chromatography column, and the functional flavor essence B rich in wolfberry polyphenols was collected. Finally, the functional flavor essence B was added back into the main wine liquid A to complete the refining and recombination.

[0040] Step S5: Aging The refined and reconstituted liquor from step S4 is aged in a constant temperature and light-proof environment at 17.5℃ for 4.5 months to obtain a multi-functional synergistic enhancement bear bile health liquor.

[0041] Comparative Examples 1-4: Comparative Example 1: Compared to Example 3, the difference lies in that the in-situ enzymatic debinding conversion in step S2 is not performed. Specifically, after step S1 is completed, the fresh bear bile that should have been added in step S2 is added together with the honey in step S3 to the compound fermented mash, followed by distillation. All other raw materials, proportions, and process parameters are the same as in Example 3.

[0042] Comparative Example 2: Compared with Example 3, the difference is that in step S1, only Saccharomyces cerevisiae is added for primary fermentation, and Lactobacillus plantarum is not added for co-fermentation. All other raw materials, proportions, and process parameters are the same as in Example 3.

[0043] Comparative Example 3: Compared to Example 3, the difference lies in that: in step S4, molecularly imprinted polymers are not used; instead, an equal volume of commercially available food-grade activated carbon is used as the adsorbent to treat the raw wine, and no subsequent elution or refilling is performed. All other raw materials, proportions, and process parameters are the same as in Example 3.

[0044] Comparative Example 4: Compared with Example 3, the difference is that this comparative example does not use fermentation, distillation, and molecular imprinting refining processes, but instead uses a traditional soaking process. Specifically, the same amount of wolfberry, fresh bear bile, and honey as in Example 3 are directly added to a grain base liquor with an alcohol content of 65% vol, and then sealed and soaked for 4.5 months in a constant temperature and light-proof environment at 17.5°C. After filtration, the finished product is obtained.

[0045] Test Example 1-2: Test Example 1: Detection method: The content of free ursodeoxycholic acid (UDCA) and the total content of wolfberry polyphenols in the final finished wines prepared in Examples 1-3 and Comparative Examples 1-4 were determined by high performance liquid chromatography (HPLC).

[0046] Instruments and columns: Agilent 1260 Infinity II high performance liquid chromatograph, equipped with a diode array detector (DAD); column: ZORBAX Eclipse Plus C18 column (4.6 mm × 250 mm, 5 µm).

[0047] Sample pretreatment: Accurately measure 5.0 mL of each sample of wine, filter it through a 0.22 µm microporous membrane, and take the filtrate as the test solution.

[0048] Determination conditions for free ursodeoxycholic acid (UDCA): Mobile phase: methanol and 0.5% glacial acetic acid aqueous solution, in a volume ratio of 85:15.

[0049] Flow rate: 1.0 mL / min.

[0050] Column temperature: 30℃.

[0051] Detection wavelength: 210nm.

[0052] Injection volume: 10µL.

[0053] Quantitative method: The external standard method was used, and a standard curve was plotted by measuring a series of UDCA standard solutions of known concentrations for quantitative calculation.

[0054] Determination conditions for total polyphenol content of wolfberry (calculated as chlorogenic acid): Mobile phase A: acetonitrile; Mobile phase B: 0.4% aqueous phosphoric acid solution.

[0055] Gradient elution program: 0-10 min, 5%-15% A; 10-25 min, 15%-25% A.

[0056] Flow rate: 0.8 mL / min.

[0057] Column temperature: 35℃.

[0058] Detection wavelength: 327nm.

[0059] Injection volume: 10µL.

[0060] Quantitative method: The external standard method was adopted. A standard curve was plotted by measuring a series of chlorogenic acid standard solutions of known concentrations. The peak areas of the main polyphenolic substances detected were summed and the total polyphenol content was calculated based on chlorogenic acid.

[0061] Test results: The results of the determination of the content of key functional components of each sample are recorded in Table 1 below.

[0062] Table 1. Results of determination of key active ingredients in each sample Table 1 shows that the content of free ursodeoxycholic acid in the finished wines prepared in Examples 1-3 was significantly higher than that in Comparative Examples 1, 2, and 4. Comparative Example 1 did not include a separate enzymatic conversion step, and Comparative Example 2 did not include *Lactobacillus plantarum*. The low level of free ursodeoxycholic acid in their finished products indicates that utilizing the complex enzyme system produced by the co-fermentation system of *Saccharomyces cerevisiae* and *Lactobacillus plantarum* to perform in-situ enzymatic hydrolysis of conjugated bile acids in fresh bear bile is a necessary process for obtaining high levels of free active ingredients.

[0063] Meanwhile, data shows that the total content of wolfberry polyphenols in the finished wines of Examples 1-3 is significantly higher than that in Comparative Examples 3 and 4. Comparative Example 3 used activated carbon to treat the raw wine, and the significant reduction in polyphenol content in its finished product indicates that non-specific adsorption materials can cause irreversible loss of these active ingredients. This technical solution uses a molecularly imprinted polymer prepared with chlorogenic acid as a template molecule to selectively capture polyphenolic components through specific recognition sites, followed by desorption and recovery using a specific eluent. This process achieves highly efficient enrichment and retention of the target components.

[0064] Based on the data in the two columns of Table 1, only the process schemes of Examples 1-3 can simultaneously ensure that both free ursodeoxycholic acid and wolfberry polyphenols are at high levels in the finished wine. This result confirms that this technical scheme systematically integrates the upstream dual-strain fermentation in-situ enzymatic conversion unit with the downstream molecular imprinting targeted anchoring refining unit, thereby solving the conversion and retention problems of different sources of functional components during the formulation process, ultimately enabling the two core functional components to coexist effectively in the finished product.

[0065] Test Example 2: Evaluation method: Ten professionally trained wine tasters conducted a double-blind evaluation of the final wines prepared in Examples 1-3 and Comparative Examples 1-4. All samples were randomly coded, and under standard tasting conditions, tasters independently scored each sample on three indicators: color, aroma harmony, and body of mouth. Each indicator was scored on a 10-point scale, with higher scores indicating better sensory performance. The average score for each sample on each indicator was calculated, and a comprehensive score was also determined.

[0066] Evaluation results: The average sensory evaluation scores for each sample are recorded in Table 2 below.

[0067] Table 2. Sensory evaluation results for each sample Table 2 shows that the finished wines prepared in Examples 1-3 scored significantly higher in aroma harmony than Comparative Examples 1, 2, and 4. In step S1, the introduction of *Lactobacillus plantarum* and *Saccharomyces cerevisiae* for synergistic fermentation resulted in a richer variety of esters and alcohols in the fermentation system. Simultaneously, the in-situ enzymatic transformation during fermentation allowed the flavor precursors in the bear bile to fully interact with the mash matrix under microbial metabolism, resulting in a more harmonious and unified complex aroma system compared to the post-addition method in Comparative Example 1. Comparative Example 2, lacking the synergistic effect of *Lactobacillus plantarum*, exhibited a single aroma profile and a lower harmony score.

[0068] The body and flavor scores of Examples 1-3 were significantly higher than those of Comparative Examples 1, 3, and 4. Comparative Example 3 used activated carbon treatment, whose non-specific adsorption caused the removal of some alcohols, esters, and polyphenols from the original wine, resulting in structural damage and a thinner mouthfeel. In this technical solution, a molecularly imprinted polymer is used in step S4. This material has selective recognition and adsorption capabilities for wolfberry polyphenols. While removing some off-flavors, the target active ingredients and flavor compounds are retained in the wine through elution and refilling, maintaining the structural integrity and body of the wine. The unconverted bound bile acids and other substances in Comparative Example 1 resulted in the lowest mouthfeel score.

[0069] The comprehensive sensory evaluation results demonstrate that this technical solution achieves harmonious sensory quality through a process design that differentiates and ultimately recombines components from different sources. The upstream dual-strain co-fermentation and in-situ enzymatic transformation steps provide the material basis for the formation of harmonious aroma characteristics. The downstream molecular imprinting targeted anchoring refining step ensures the fullness of the wine while preserving key flavor components. This systematic process combination results in a final product that outperforms its counterparts in terms of color, aroma, and taste, confirming the effectiveness of this technical solution in improving the sensory quality of the product.

Claims

1. A multi-effect synergistically enhanced bear gall health wine compound preparation process, characterized in that, The method comprises the following steps: S1, mix sorghum and corn which are mixed and crushed and passed through a 20-40 mesh sieve with wolfberry in whole particle form, add brewing yeast for primary fermentation, then add lactobacillus plantarum for synergistic fermentation to obtain a compound fermented wine mash; S2, add fresh bear bile to the compound fermented wine mash obtained in step S1 for in-situ enzymatic debonding conversion; S3, add honey to the compound fermented wine mash after conversion in step S2 for flavor coupling, then distill to collect base liquor; S4, pass the base liquor obtained in step S3 through a chromatographic column filled with molecularly imprinted polymers, collect the main liquor A flowing out, then elute the molecularly imprinted polymers with an eluent, collect the efficacy and flavor essence B rich in wolfberry polyphenols, finally add the efficacy and flavor essence B back to the main liquor A to complete refining and recombination to obtain refined and recombined liquor; S5, age the refined and recombined liquor obtained in step S4 in a constant temperature and light-proof environment at 15-20℃ for 3-6 months to obtain multi-efficacy synergistically enhanced bear bile health wine.

2. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 1, characterized in that, In step S1, the dry weight ratio of sorghum, corn and wolfberry is 6:3:

1.

3. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 1, characterized in that, In step S1, the adding amount of brewing yeast is 0.3-0.5% of the total dry weight of raw materials, and the adding amount of lactobacillus plantarum is 0.1-0.2% of the total dry weight of raw materials; wherein the total dry weight of raw materials is the sum of the dry weights of sorghum, corn and wolfberry in step S1.

4. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 1, characterized in that, In step S1, the operation of adding lactobacillus plantarum is performed when the temperature of the primary fermentation product rises to 35-38℃.

5. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 1, characterized in that, In step S2, the adding amount of fresh bear bile is 0.5-1.5% of the total dry weight of raw materials, the temperature of in-situ enzymatic debonding conversion is 30-35℃, and the time is 48-72 hours.

6. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 1, characterized in that, In step S3, the adding amount of honey is 2.0-3.0% of the total dry weight of raw materials, and the intermediate section base liquor with an alcohol content of 60-70%vol is collected in the distillation step.

7. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 1, characterized in that, In step S4, the preparation method of the molecularly imprinted polymers is as follows: chloroformic acid is used as a template molecule, 4-vinylpyridine is used as a functional monomer, ethylene glycol dimethacrylate is used as a crosslinking agent, and azobisisobutyronitrile is used as an initiator to perform a thermal initiation polymerization reaction, wherein the molar ratio of the template molecule, the functional monomer and the crosslinking agent is 1:4:20 to obtain the molecularly imprinted polymers.

8. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 1, characterized in that, In step S4, the eluent is an ethanol aqueous solution with an ethanol volume fraction of 70-80% and containing 0.5-1.0% by volume of glacial acetic acid.

9. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 7, characterized in that, The thermal initiation polymerization reaction is performed at a constant temperature of 60-70℃ for 20-28 hours.

10. The multi-function synergistically enhanced bear gall health wine compound preparation process according to claim 7, characterized in that, The preparation method of the molecularly imprinted polymers further comprises: after the thermal initiation polymerization reaction, a Soxhlet extraction is performed on the polymer using a mixed solution of methanol and glacial acetic acid with a volume ratio of 9:1 to remove the template molecule, thereby obtaining the molecularly imprinted polymers with specific recognition cavities.

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