Garlic powder with core-shell structure as well as preparation method and application of garlic powder
By combining enzymatic hydrolysis and specific ultrafiltration membrane purification with a core-shell structure, a method for preparing garlic powder was developed, which solved the problems of insufficient stability and targeting of active ingredients in garlic powder during NASH treatment. This method achieved targeted sustained release in liver tissue, improved the therapeutic effect, and reduced the risk of systemic adverse reactions.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- 物生生物科技(北京)有限公司
- Filing Date
- 2026-01-29
- Publication Date
- 2026-04-17
AI Technical Summary
Existing garlic powder treatments for non-alcoholic steatohepatitis (NASH) suffer from poor stability of active ingredients, lack of targeting and sustained-release effects, resulting in low bioavailability and potentially increased risk of systemic adverse reactions. Existing core-shell structured materials are not stable enough in acidic environments, and the preparation process may cause residue problems.
The active ingredients of garlic were purified by compound enzymatic hydrolysis and ultrafiltration membrane with a specific molecular weight cutoff. The core-shell structure was formed by combining pullulan, galactose, and polycarboxylic acids. The core-shell structured garlic powder with a particle size of 1-2 μm and a PDI between 0.3 and 0.35 was prepared by vacuum freeze-drying technology, achieving a sustained-release effect with a cumulative release rate of <15% in gastric juice over 2 hours and >70% in intestinal juice over 12 hours.
It improves the stability and targeting of garlic powder, avoids the degradation of active ingredients in the stomach, and achieves targeted sustained release in the intestine, effectively treating non-alcoholic steatohepatitis.
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Abstract
Description
Technical Field
[0001] This application relates to the field of pharmaceutical technology, specifically to a core-shell structured garlic powder, its preparation method, and its application. Background Technology
[0002] Nonalcoholic fatty liver disease (NAFLD) is one of the most prevalent chronic liver diseases worldwide, characterized by metabolic stress-induced liver injury caused by excessive fat deposition in hepatocytes. Its core pathological features include hepatocyte steatosis, inflammatory infiltration, ballooning degeneration of hepatocytes, and liver fibrosis. It is also highly comorbid with metabolic-related diseases such as type 2 diabetes, metabolic syndrome, and obesity. Nonalcoholic steatohepatitis (NASH), a severe progressive form of NAFLD, is characterized by significant inflammatory response, hepatocyte damage, and liver fibrosis in addition to steatosis.
[0003] The pathogenesis of NASH is complex, involving the interaction of multiple factors such as environment, diet, metabolism, and genetics. Its core pathological processes include insulin resistance, lipid metabolism disorders, oxidative stress, inflammatory responses, and hepatocyte ferroptosis. Currently, there are no specific drugs for treating NASH in clinical practice; treatment mainly relies on lifestyle interventions, but these have limited effectiveness and poor patient adherence. Therefore, developing safe and effective drugs for the treatment of NASH has significant clinical value and social implications.
[0004] Garlic, a traditional food and medicine source, contains active ingredients such as allicin and sulfides, which possess various pharmacological activities including anti-inflammatory, antioxidant, and metabolic regulation, showing potential application prospects in the prevention and treatment of liver diseases. However, natural garlic or conventional garlic powder has significant limitations: on the one hand, the active ingredients such as allicin are unstable and easily oxidized and degraded by factors such as temperature, oxygen, and light, resulting in low bioavailability; on the other hand, the active ingredients in conventional garlic powder lack targeting and are widely distributed in the body, making it difficult to accumulate in liver tissue, which affects the therapeutic effect and may increase the risk of systemic adverse reactions. In addition, the release rate of garlic's active ingredients is difficult to control, and they are easily released rapidly and destroyed in the gastrointestinal tract, further limiting its application in the treatment of NASH.
[0005] To address these issues, researchers have attempted to improve the delivery efficiency of garlic's active ingredients through formulation technology. Core-shell structures, as a novel drug delivery system, utilize a core carrier to load the active ingredient, while the outer shell provides protection, targeting, and sustained-release functions, and have garnered widespread attention in the pharmaceutical field. Currently, existing core-shell structures for garlic formulations primarily use sodium alginate and chitosan as shell materials. While these materials possess some biocompatibility, they suffer from the following drawbacks: the gel strength of sodium alginate is easily affected by ionic strength, leading to insufficient shell structure stability; chitosan has poor water solubility and is prone to protonation in acidic environments, potentially causing gastrointestinal discomfort, thus limiting its application in food-grade or high-safety drugs. Furthermore, existing core-shell structures often feature single-function shell designs, lacking comprehensive consideration of active ingredient stability, targeted delivery, sustained-release regulation, and synergistic therapy. Additionally, some preparation processes utilize chemical cross-linking agents, posing residual risks and failing to meet the requirements for high-safety drugs.
[0006] Therefore, how to provide a core-shell structured garlic powder that can treat non-alcoholic steatohepatitis has become a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention
[0007] The purpose of this application is to overcome the shortcomings of the existing technology and provide a core-shell structured garlic powder, its preparation method and application. The core-shell structured garlic powder of this application can effectively treat non-alcoholic steatohepatitis.
[0008] To achieve the above objectives, the first aspect of this application provides a method for preparing core-shell structured garlic powder, comprising the following steps: (1) Grind the garlic and water together to obtain a paste; (2) Add the compound enzyme to the slurry and hydrolyze it to obtain the hydrolysate; (3) The enzymatic hydrolysate is first ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 1000 Da, and then ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 100 Da to obtain the ultrafiltrate. (4) The ultrafiltrate was freeze-dried under vacuum to obtain garlic powder; (5) Add pullulan, galactose, and polycarboxylic acid to water, stir well, then add branched starch, chitosan and maltodextrin, stir well to obtain shell liquid; (6) Add garlic powder to the shell liquid, stir evenly with ultrasound, and freeze dry under vacuum to obtain core-shell structured garlic powder.
[0009] This application involves first pulping the garlic to promote the full release of active garlic components from the cells into the pulp, providing sufficient reaction substrate for subsequent enzymatic hydrolysis; then, through enzymatic hydrolysis with a complex enzyme, promoting the complete disruption of garlic cells and the full release of active components that treat non-alcoholic fatty liver disease; finally, through bipolar ultrafiltration using an ultrafiltration membrane with a specific molecular weight cutoff, retaining small-molecule active components, effectively removing impurities, and effectively improving purity, making it more conducive to human absorption; and then, through vacuum freeze-drying, improving the retention rate of active components, preventing oxidation and decomposition of active components, and effectively controlling the particle size and PDI of the garlic powder, preventing the garlic powder from absorbing moisture or agglomerating. The obtained garlic powder is then coated with shell liquid to obtain core-shell structured garlic powder. The average particle size of the core-shell structured garlic powder is between 1 and 2 μm, and the PDI is between 0.3 and 0.35, exhibiting excellent stability. At the same time, the core-shell structured garlic powder has a good sustained-release effect. In simulated gastric juice, the cumulative release rate should be <15% after 2 hours, reflecting the barrier effect of the shell layer on gastric acid and preventing the degradation of active ingredients in the stomach. In simulated intestinal juice, the cumulative release rate should be >70% after 12 hours, achieving targeted sustained release of active ingredients in the intestine, which can effectively treat non-alcoholic steatohepatitis.
[0010] As an embodiment of this application, the mass ratio of garlic to water is 1:(2~10).
[0011] As an embodiment of this application, the mass ratio of the compound enzyme to the slurry is (0.008~0.02):1.
[0012] As an embodiment of this application, the complex enzyme comprises cellulase, neutral protease and papain in a mass ratio of 1:(0.5~1.2):(0.5~0.8).
[0013] This application uses cellulase, neutral protease, and papain in a mass ratio of 1:(0.5~1.2):(0.5~0.8) as a complex enzyme to target and degrade the cellulose skeleton of garlic cell walls, break down the cell barrier, and hydrolyze large molecular proteins and poorly soluble proteins in garlic under mild conditions. The proteins are hydrolyzed into small molecular peptides and amino acids, promoting the full release of active ingredients that have therapeutic effects on non-alcoholic steatohepatitis (NAH), and effectively improving the therapeutic effect on NHA.
[0014] As an embodiment of this application, the enzymatic hydrolysis temperature is 45~50℃ and the time is 0.5~2h.
[0015] As an implementation scheme of this application, the vacuum freeze-drying in step (4) specifically involves: first drying at a vacuum of 28~32 Pa and a temperature of -10~-5℃ for 0.5~2 h; then drying at a vacuum of 60~65 Pa and a temperature of -10~-5℃ for 1~3 h; then drying at a vacuum of 12~16 Pa and a temperature of -2~2℃ for 0.5~1 h; and finally drying at a vacuum of 8~12 Pa and a temperature of 18~22℃ for 1~3 h. This application, through specific vacuum freeze-drying, can improve the retention rate of active ingredients, avoid oxidation and decomposition of active ingredients, and effectively control the particle size and PDI of garlic powder, preventing the garlic powder from absorbing moisture or agglomerating.
[0016] As an embodiment of this application, the mass ratio of pullulan, galactose, polycarboxylic acid, water, amylopectin, chitosan and hydroxypropyl-β-cyclodextrin is (2~3):1:(0.08~0.12):(15~30):(0.1~0.2):(0.8~1.5).
[0017] This application uses a mass ratio of (2~3):1:(0.08~0.12):(15~30):(0.1~0.2):(0.8~1.5) as the shell coating material, and pullulan and chitosan as the main film-forming materials. These materials possess appropriate water solubility and film-forming properties, forming a dense coating layer. The galactose enhances the targeting effect and strengthens the affinity between the shell and the core. The polycarboxylic acid undergoes an esterification reaction with chitosan, improving the stability and water resistance of the shell layer. The branched starch and hydroxypropyl... -β-cyclodextrin improves the flowability and structural stability of garlic powder. Under specific proportions of the above-mentioned raw materials, a shell-shell liquid with suitable viscosity and uniform dispersion is obtained, which has good film-forming and coating capabilities and can promote the formation of garlic powder with a core-shell structure that has excellent sustained-release effect and stability. The average particle size of the core-shell structured garlic powder is between 1 and 2 μm, and the PDI is between 0.3 and 0.35, exhibiting excellent stability. At the same time, the core-shell structured garlic powder has a good sustained-release effect. In simulated gastric juice, the cumulative release rate should be <15% after 2 hours, reflecting the barrier effect of the shell layer on gastric acid and preventing the degradation of active ingredients in the stomach; in simulated intestinal juice, the cumulative release rate should be >70% after 12 hours.
[0018] As an embodiment of this application, the mass ratio of garlic powder to shell liquid is 1:(3~4).
[0019] As an implementation scheme of this application, the vacuum freeze drying in step (6) specifically involves: first drying at a vacuum degree of 32~38 Pa and a temperature of -16~-14℃ for 0.5~2h; then drying at a vacuum degree of 45~50 Pa and a temperature of -8~-5℃ for 1~2h; then drying at a vacuum degree of 12~16 Pa and a temperature of -2~2℃ for 0.5~2h; and finally drying at a vacuum degree of 8~12 Pa and a temperature of 22~26℃ for 1~3h.
[0020] As an embodiment of this application, the polycarboxylic acid includes at least one of malic acid and citric acid.
[0021] As an embodiment of this application, the power of the ultrasound is 200~800W, for example, it can be 200W, 300W, 400W, 500W, 600W, 700W, 800W or any two of these values.
[0022] The second aspect of this application provides a core-shell structured garlic powder, prepared using the preparation method described above.
[0023] A third aspect of this application provides the use of core-shell structured garlic powder in the preparation of a medicament for treating non-alcoholic steatohepatitis.
[0024] The beneficial effects of this application are as follows: First, garlic is pulped to promote the full release of active garlic components from the cells into the pulp, providing sufficient reaction substrate for subsequent enzymatic hydrolysis. Second, enzymatic hydrolysis with a complex enzyme promotes the complete disruption of garlic cells, facilitating the full release of active components that treat non-alcoholic fatty liver disease. Third, ultrafiltration using a specific molecular weight cutoff membrane retains small-molecule active components, effectively removes impurities, and significantly improves purity, making it more suitable for human absorption. Fourth, vacuum freeze-drying further enhances the retention rate of active components, prevents oxidation and decomposition, and effectively controls the particle size and PDI of the garlic powder, preventing moisture absorption or agglomeration. The obtained garlic powder is then coated with shell liquid to obtain core-shell structured garlic powder. The average particle size of the core-shell structured garlic powder is between 1 and 2 μm, and the PDI is between 0.3 and 0.35, exhibiting excellent stability. At the same time, the core-shell structured garlic powder has a good sustained-release effect. In simulated gastric juice, the cumulative release rate should be <15% after 2 hours, reflecting the barrier effect of the shell layer on gastric acid and preventing the degradation of active ingredients in the stomach. In simulated intestinal juice, the cumulative release rate should be >70% after 12 hours, achieving targeted sustained release of active ingredients in the intestine, which can effectively treat non-alcoholic steatohepatitis. Detailed Implementation
[0025] In this application, the technical features described in an open-ended manner include both closed technical solutions consisting of the listed features and open technical solutions that include the listed features.
[0026] In this application, numerical ranges are referred to as continuous unless otherwise specified, and include the minimum and maximum values of the range, as well as every value between the minimum and maximum values. Furthermore, when the range refers to integers, it includes every integer between the minimum and maximum values of the range. Additionally, when multiple ranges are provided to describe a feature or characteristic, the ranges may be merged. In other words, unless otherwise specified, all ranges disclosed herein should be understood to include any and all subranges to which they are incorporated.
[0027] In this application, there are no particular restrictions on the specific dispersion and mixing methods.
[0028] Unless otherwise specified, all components, raw materials, or instruments used in the embodiments and comparative examples of this application are commercially available, and the components and raw materials used in each parallel experiment are the same.
[0029] The amylopectin in this application is derived from Beijing Solarbio.
[0030] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of this application, not all embodiments. Based on the embodiments in this application, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of this application.
[0031] Example 1 A method for preparing garlic powder with a core-shell structure includes the following steps: (1) Take fresh garlic, remove rotten and broken garlic cloves and impurities, rinse the surface mud and residual dirt with running deionized water, dry at 60℃ for 10h, add water to the dried garlic and pulp it at 8000rpm for 5min, filter it to remove uncrushed garlic and residue, and obtain pulp. (2) Add the compound enzyme to the slurry and hydrolyze it at 46°C for 1.5 h to obtain the hydrolysate; the mass ratio of the compound enzyme to the slurry is 0.018:1, and the compound enzyme includes cellulase, neutral protease and papain in a mass ratio of 1:0.8:0.6.
[0032] (3) The enzymatic hydrolysate is first ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 1000 Da, and then ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 100 Da to obtain the ultrafiltrate. (4) The ultrafiltrate was added to a freeze dryer and dried for 1 hour at a vacuum of 30 Pa and a temperature of -8 °C; then dried for 2 hours at a vacuum of 62 Pa and a temperature of -8 °C; then dried for 0.8 hours at a vacuum of 15 Pa and a temperature of 0 °C; and finally dried for 2 hours at a vacuum of 10 Pa and a temperature of 20 °C to obtain garlic powder. (5) Add pullulan, galactose, and malic acid to water and stir at 100 rpm for 0.5 h at 45 °C. Then add amylopectin, chitosan, and maltodextrin and stir at 100 rpm for 1 h at 45 °C to obtain a shell liquid. The mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin is 2.5:1:0.1:20:0.16:1.
[0033] (6) Add garlic powder to shell liquid (the mass ratio of garlic powder to shell liquid is 1:4), stir at 200 rpm for 30 min under 500W ultrasonication, and add the resulting material to a freeze dryer. First, dry at a vacuum of 35 Pa and a temperature of -15℃ for 1 h; then dry at a vacuum of 50 Pa and a temperature of -6℃ for 1.5 h; then dry at a vacuum of 15 Pa and a temperature of 0℃ for 1 h; finally dry at a vacuum of 10 Pa and a temperature of 25℃ for 2 h to obtain core-shell structured garlic powder.
[0034] Example 2 The difference between Example 2 and Example 1 is that the composition of the complex enzyme is different, but everything else is the same.
[0035] The complex enzyme in Example 2 comprises cellulase, neutral protease, and papain in a mass ratio of 1:0.5:0.5.
[0036] Example 3 The difference between Example 3 and Example 1 is that the composition of the complex enzyme is different, but everything else is the same.
[0037] The complex enzyme in Example 3 comprises cellulase, neutral protease, and papain in a mass ratio of 1:1.2:0.8.
[0038] Example 4 Example 4 differs from Example 1 in that the mass ratios of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin are different, while all other components are the same.
[0039] In Example 4, the mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin was 2:1:0.08:20:0.1:0.8.
[0040] Example 5 Example 5 differs from Example 1 in that the mass ratios of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin are different, while all other components are the same.
[0041] In Example 5, the mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin was 3:1:0.12:20:0.2:1.5.
[0042] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that the preparation methods of the core-shell structured garlic powder are different.
[0043] A method for preparing garlic powder with a core-shell structure includes the following steps: (1) Take fresh garlic, remove rotten and broken garlic cloves and impurities, rinse the surface mud and residual dirt with running deionized water, dry at 60℃ for 10h, add water to the dried garlic and pulp it at 8000rpm for 5min, filter it to remove uncrushed garlic and residue, and obtain pulp. (2) Add the compound enzyme to the slurry and hydrolyze it at 46°C for 1.5 h to obtain the hydrolysate; the mass ratio of the compound enzyme to the slurry is 0.018:1, and the compound enzyme includes cellulase, neutral protease and papain in a mass ratio of 1:0.8:0.6.
[0044] (3) The ultrafiltrate was added to a freeze dryer and dried for 1 hour at a vacuum of 30 Pa and a temperature of -8 °C; then dried for 2 hours at a vacuum of 62 Pa and a temperature of -8 °C; then dried for 0.8 hours at a vacuum of 15 Pa and a temperature of 0 °C; and finally dried for 2 hours at a vacuum of 10 Pa and a temperature of 20 °C to obtain garlic powder. (4) Add pullulan, galactose, and malic acid to water and stir at 100 rpm for 0.5 h at 45 °C. Then add amylopectin, chitosan, and maltodextrin and stir at 100 rpm for 1 h at 45 °C to obtain a shell liquid. The mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin is 2.5:1:0.1:20:0.16:1.
[0045] (5) Add garlic powder to shell liquid (the mass ratio of garlic powder to shell liquid is 1:4), stir at 200 rpm for 30 min under 500W ultrasonication, and add the resulting material to a freeze dryer. First, dry at a vacuum of 35 Pa and a temperature of -15℃ for 1 h; then dry at a vacuum of 50 Pa and a temperature of -6℃ for 1.5 h; then dry at a vacuum of 15 Pa and a temperature of 0℃ for 1 h; finally dry at a vacuum of 10 Pa and a temperature of 25℃ for 2 h to obtain core-shell structured garlic powder.
[0046] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that the preparation method of the core-shell structured garlic powder is different.
[0047] A method for preparing garlic powder with a core-shell structure includes the following steps: (1) Take fresh garlic, remove rotten and broken garlic cloves and impurities, rinse the surface mud and residual dirt with running deionized water, dry at 60℃ for 10h, add water to the dried garlic and pulp it at 8000rpm for 5min, filter it to remove uncrushed garlic and residue, and obtain pulp. (2) Add the compound enzyme to the slurry and hydrolyze it at 46°C for 1.5 h to obtain the hydrolysate; the mass ratio of the compound enzyme to the slurry is 0.018:1, and the compound enzyme includes cellulase, pectinase and trypsin in a mass ratio of 1:0.8:0.6.
[0048] (3) The enzymatic hydrolysate is first ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 1000 Da, and then ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 100 Da to obtain the ultrafiltrate. (4) The ultrafiltrate was added to a freeze dryer and dried for 1 hour at a vacuum of 30 Pa and a temperature of -8 °C; then dried for 2 hours at a vacuum of 62 Pa and a temperature of -8 °C; then dried for 0.8 hours at a vacuum of 15 Pa and a temperature of 0 °C; and finally dried for 2 hours at a vacuum of 10 Pa and a temperature of 20 °C to obtain garlic powder. (5) Add pullulan, galactose, and malic acid to water and stir at 100 rpm for 0.5 h at 45 °C. Then add amylopectin, chitosan, and maltodextrin and stir at 100 rpm for 1 h at 45 °C to obtain a shell liquid. The mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin is 2.5:1:0.1:20:0.16:1.
[0049] (6) Add garlic powder to shell liquid (the mass ratio of garlic powder to shell liquid is 1:4), stir at 200 rpm for 30 min under 500W ultrasonication, and add the resulting material to a freeze dryer. First, dry at a vacuum of 35 Pa and a temperature of -15℃ for 1 h; then dry at a vacuum of 50 Pa and a temperature of -6℃ for 1.5 h; then dry at a vacuum of 15 Pa and a temperature of 0℃ for 1 h; finally dry at a vacuum of 10 Pa and a temperature of 25℃ for 2 h to obtain core-shell structured garlic powder.
[0050] Comparative Example 3 The difference between Comparative Example 3 and Example 1 is that the preparation method of the core-shell structured garlic powder is different.
[0051] A method for preparing garlic powder with a core-shell structure includes the following steps: (1) Take fresh garlic, remove rotten and broken garlic cloves and impurities, rinse the surface mud and residual dirt with running deionized water, dry at 60℃ for 10h, add water to the dried garlic and pulp it at 8000rpm for 5min, filter it to remove uncrushed garlic and residue, and obtain pulp. (2) Add the compound enzyme to the slurry and hydrolyze it at 46°C for 1.5 h to obtain the hydrolysate; the mass ratio of the compound enzyme to the slurry is 0.018:1, and the compound enzyme includes pectinase, β-glucanase and neutral protease in a mass ratio of 1:0.8:0.6.
[0052] (3) The enzymatic hydrolysate is first ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 1000 Da, and then ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 100 Da to obtain the ultrafiltrate. (4) The ultrafiltrate was added to a freeze dryer and dried for 1 hour at a vacuum of 30 Pa and a temperature of -8 °C; then dried for 2 hours at a vacuum of 62 Pa and a temperature of -8 °C; then dried for 0.8 hours at a vacuum of 15 Pa and a temperature of 0 °C; and finally dried for 2 hours at a vacuum of 10 Pa and a temperature of 20 °C to obtain garlic powder. (5) Add pullulan, galactose, and malic acid to water and stir at 100 rpm for 0.5 h at 45 °C. Then add amylopectin, chitosan, and maltodextrin and stir at 100 rpm for 1 h at 45 °C to obtain a shell liquid. The mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin is 2.5:1:0.1:20:0.16:1.
[0053] (6) Add garlic powder to shell liquid (the mass ratio of garlic powder to shell liquid is 1:4), stir at 200 rpm for 30 min under 500W ultrasonication, and add the resulting material to a freeze dryer. First, dry at a vacuum of 35 Pa and a temperature of -15℃ for 1 h; then dry at a vacuum of 50 Pa and a temperature of -6℃ for 1.5 h; then dry at a vacuum of 15 Pa and a temperature of 0℃ for 1 h; finally dry at a vacuum of 10 Pa and a temperature of 25℃ for 2 h to obtain core-shell structured garlic powder.
[0054] Comparative Example 4 The difference between Comparative Example 4 and Example 1 is that the composition of the complex enzyme is different, but everything else is the same.
[0055] The complex enzyme in Comparative Example 4 consisted of cellulase, neutral protease, and papain in a mass ratio of 1:0.25:0.25.
[0056] Comparative Example 5 The difference between Comparative Example 5 and Example 1 is that the composition of the complex enzyme is different, but everything else is the same.
[0057] The complex enzyme in Comparative Example 5 consisted of cellulase, neutral protease, and papain in a mass ratio of 1:2:2.
[0058] Comparative Example 6 The difference between Comparative Example 6 and Example 1 is that the mass ratios of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin are different in Comparative Example 6, while all other components are the same.
[0059] The mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan and hydroxypropyl-β-cyclodextrin in Comparative Example 6 was 1:1:0.05:20:0.05:0.5.
[0060] Comparative Example 7 The difference between Comparative Example 7 and Example 1 is that the mass ratios of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin are different in Comparative Example 7, while all other components are the same.
[0061] The mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan and hydroxypropyl-β-cyclodextrin in Comparative Example 7 was 4:1:0.2:20:0.4:2.
[0062] Comparative Example 8 The difference between Comparative Example 8 and Example 1 is that an equal amount of sodium alginate was used to replace pullulan and galactose, while all other aspects were the same.
[0063] A method for preparing garlic powder with a core-shell structure includes the following steps: (1) Take fresh garlic, remove rotten and broken garlic cloves and impurities, rinse the surface mud and residual dirt with running deionized water, dry at 60℃ for 10h, add water to the dried garlic and pulp it at 8000rpm for 5min, filter it to remove uncrushed garlic and residue, and obtain pulp. (2) Add the compound enzyme to the slurry and hydrolyze it at 46°C for 1.5 h to obtain the hydrolysate; the mass ratio of the compound enzyme to the slurry is 0.018:1, and the compound enzyme includes cellulase, neutral protease and papain in a mass ratio of 1:0.8:0.6.
[0064] (3) The enzymatic hydrolysate is first ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 1000 Da, and then ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 100 Da to obtain the ultrafiltrate. (4) The ultrafiltrate was added to a freeze dryer and dried for 1 hour at a vacuum of 30 Pa and a temperature of -8 °C; then dried for 2 hours at a vacuum of 62 Pa and a temperature of -8 °C; then dried for 0.8 hours at a vacuum of 15 Pa and a temperature of 0 °C; and finally dried for 2 hours at a vacuum of 10 Pa and a temperature of 20 °C to obtain garlic powder. (5) Sodium alginate and malic acid are added to water and stirred at 100 rpm for 0.5 h at 45 °C. Then, amylopectin, chitosan and maltodextrin are added and stirred at 100 rpm for 1 h at 45 °C to obtain shell liquid. The mass ratio of sodium alginate, malic acid, water, amylopectin, chitosan and hydroxypropyl-β-cyclodextrin is 3.5:0.1:20:0.16:1.
[0065] (6) Add garlic powder to shell liquid (the mass ratio of garlic powder to shell liquid is 1:4), stir at 200 rpm for 30 min under 500W ultrasonication, and add the resulting material to a freeze dryer. First, dry at a vacuum of 35 Pa and a temperature of -15℃ for 1 h; then dry at a vacuum of 50 Pa and a temperature of -6℃ for 1.5 h; then dry at a vacuum of 15 Pa and a temperature of 0℃ for 1 h; finally dry at a vacuum of 10 Pa and a temperature of 25℃ for 2 h to obtain core-shell structured garlic powder.
[0066] Comparative Example 9 The difference between Comparative Example 9 and Example 1 is that the preparation method of the core-shell structured garlic powder in Comparative Example 9 is different from that in Example 1.
[0067] A method for preparing garlic powder with a core-shell structure includes the following steps: (1) Take fresh garlic, remove rotten and broken garlic cloves and impurities, rinse the surface mud and residual dirt with running deionized water, dry at 60℃ for 10h, add water to the dried garlic and pulp it at 8000rpm for 5min, filter it to remove uncrushed garlic and residue, and obtain pulp. (2) Add the compound enzyme to the slurry and hydrolyze it at 46°C for 1.5 h to obtain the hydrolysate; the mass ratio of the compound enzyme to the slurry is 0.018:1, and the compound enzyme includes cellulase, neutral protease and papain in a mass ratio of 1:0.8:0.6.
[0068] (3) The enzymatic hydrolysate is first ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 1000 Da, and then ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 100 Da to obtain the ultrafiltrate. (4) The ultrafiltrate was added to a freeze dryer and dried for 1 hour at a vacuum of 30 Pa and a temperature of -8 °C; then dried for 2 hours at a vacuum of 62 Pa and a temperature of -8 °C; then dried for 0.8 hours at a vacuum of 15 Pa and a temperature of 0 °C; and finally dried for 2 hours at a vacuum of 10 Pa and a temperature of 20 °C to obtain garlic powder. (5) Add pullulan, galactose, and malic acid to water and stir at 100 rpm for 0.5 h at 45 °C. Then add amylopectin, chitosan, and maltodextrin and stir at 100 rpm for 1 h at 45 °C to obtain a shell liquid. The mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin is 2.5:1:0.1:20:0.16:1.
[0069] (6) Add garlic powder to shell liquid (the mass ratio of garlic powder to shell liquid is 1:4), stir at 200 rpm for 30 min under 500W ultrasound, and dry the product at 75℃ for 12 h to obtain core-shell structured garlic powder.
[0070] Test case 1. To investigate the effect of the preparation process on the stability of core-shell garlic powder, the core-shell garlic powder from Examples 1, 4, 5, and Comparative Examples 6-9 were diluted 10 times with water for injection. A Malvern Zetasizer Nano ZS90 nanoparticle size analyzer was used, with the detection temperature set at 25°C and the equilibration time at 120 s. The average particle size (Z-average particle size) and polydispersity index (PDI) of the liposomes were determined using dynamic light scattering (DLS). Each sample was measured in triplicate, and the average value was taken as the final result.
[0071] Table 1 As can be seen from Table 1, the core-shell structured garlic powder described in this application has the characteristics of small particle size and uniform PDI distribution. The core-shell structured garlic powder prepared in this application has an average particle size of 1~2μm and a PDI of 0.3~0.35, exhibiting excellent stability.
[0072] Comparing Examples 1, 4, and 5 with Comparative Examples 6 and 7, it can be seen that the mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin has a significant impact on the average particle size and PDI. This application obtained a core-shell structured garlic powder with an average particle size between 1 and 2 μm and a PDI between 0.3 and 0.35 by controlling the mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin to (2~3):1:(0.08~0.12):(15~30):(0.1~0.2):(0.8~1.5).
[0073] Comparing Example 1 and Comparative Example 8, it can be seen that different shell materials have a significant impact on average particle size and PDI. This application uses pullulan, galactose, malic acid, water, amylopectin, chitosan and hydroxypropyl-β-cyclodextrin as shell materials. Compared with conventional combined shell materials such as sodium alginate and chitosan, it can significantly improve the average particle size and PDI, and effectively improve the stability of core-shell structure garlic powder.
[0074] Comparing Example 1 and Comparative Example 9, it can be seen that after mixing garlic powder and shell liquid, this application can significantly improve the average particle size and PDI by adopting a specific vacuum freeze-drying process, effectively improving the stability of core-shell structured garlic powder.
[0075] 2. To investigate the effect of the preparation process on the sustained-release effect of core-shell structured garlic powder, the sustained-release effect was tested in simulated gastric juice (pH 2.0, SGJ) and simulated intestinal juice (pH 7.0, SIF). The cumulative release rate of core-shell structured garlic powder in simulated gastric juice should be <15% after 2 hours, reflecting the barrier effect of the shell on gastric acid and preventing the degradation of active ingredients in the stomach; the cumulative release rate in simulated intestinal juice should be >70% after 12 hours, achieving targeted sustained release of active ingredients in the intestine.
[0076] The garlic powder with a core-shell structure from Examples 1, 4, 5, and Comparative Examples 6-9 were prepared into a 20% aqueous solution, and the content of active ingredients (allicin and alliin) was determined by HPLC and denoted as m1.
[0077] The garlic powder with a core-shell structure from Examples 1, 4, 5, and Comparative Examples 6-9 were prepared into a 20% aqueous solution and placed in dialysis bags, which were then placed in simulated intestinal fluid and simulated gastric fluid, respectively. After 2 hours in simulated gastric fluid, the content of active ingredients (allicin and alliin) was determined by HPLC and recorded as m2. After 12 hours in simulated intestinal fluid, the content of active ingredients (allicin and alliin) was determined by HPLC and recorded as m3. The sustained-release rate was calculated.
[0078] Simulated intestinal fluid sustained release rate = (m3 / m1) * 100%.
[0079] Simulated gastric juice sustained release rate = (m2 / m1) * 100%.
[0080] Table 2 As can be seen from Table 2, the cumulative release rate of the core-shell structure described in this application in simulated gastric juice should be <15% in 2 hours, reflecting the barrier effect of the shell on gastric acid and preventing the degradation of active ingredients in the stomach; the cumulative release rate in simulated intestinal juice should be >70% in 12 hours, achieving targeted sustained release of active ingredients in the intestine.
[0081] Comparing Examples 1, 4, and 5 with Comparative Examples 6 and 7, it can be seen that the mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin has a significant impact on the sustained-release effect. This application can effectively improve the sustained-release effect of core-shell structured garlic powder in the intestine and stomach by controlling the mass ratio of pullulan, galactose, malic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin to (2~3):1:(0.08~0.12):(15~30):(0.1~0.2):(0.8~1.5).
[0082] Comparing Example 1 and Comparative Example 8, it can be seen that different shell materials have a significant impact on the sustained-release effect of core-shell structured garlic powder. This application uses pullulan, galactose, malic acid, water, amylopectin, chitosan and hydroxypropyl-β-cyclodextrin as shell materials, which can significantly improve the sustained-release effect of core-shell structured garlic powder in the intestine and stomach compared with conventional combination shell materials such as sodium alginate and chitosan.
[0083] 3. To investigate the effect of preparation process on the therapeutic efficacy of core-shell garlic powder in treating NASH, C57BL / 6J mice weighing 22±2g were selected. The barrier environment was set at room temperature (22±1)℃, relative humidity 30~60%, 12h light / 12h dark, and the animals had free access to food and water. NASH was induced by feeding them MCD model diet for 4 weeks. The successfully modeled mice were randomly divided into control group, model group, positive control group and experimental group, with 8 mice in each group. The experimental group was administered core-shell garlic powder (8g / kg / day) of Examples 1-3 and Comparative Examples 1-5 by gavage. The control group and model group were given the corresponding physiological saline. The positive control group was given 8wt% pioglitazone hydrochloride solution. After 5 weeks, various indicators of the mice were detected.
[0084] Table 3 As can be seen from Table 3, the core-shell structured garlic powder described in this application can significantly improve liver TG and TC, as well as liver index, TBA and AST, and has a significant therapeutic effect on non-alcoholic steatohepatitis.
[0085] Comparing Examples 1-3 with Comparative Examples 2-5, the types of enzymes mentioned have a significant impact on the therapeutic effect of non-alcoholic steatohepatitis (NAH). Different types of enzymes and enzyme mass ratios both affect the therapeutic effect of NHA. This application effectively improves the therapeutic effect of NHA by using a complex enzyme of cellulase, neutral protease, and papain in a mass ratio of 1:(0.5-1.2):(0.5-0.8).
[0086] Comparing Example 1 with Comparative Example 1, it can be seen that this application effectively improves the treatment effect of non-alcoholic steatohepatitis by employing a specific ultrafiltration step.
[0087] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this application and are not intended to limit the scope of protection of this application. Although this application has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications or equivalent substitutions can be made to the technical solutions of this application without departing from the substance and scope of the technical solutions of this application.
Claims
1. A method for preparing core-shell structured garlic powder, characterized in that, Includes the following steps: (1) Grind the garlic and water together to obtain a paste; (2) Add the compound enzyme to the slurry and hydrolyze it to obtain the hydrolysate; (3) The enzymatic hydrolysate is first ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 1000 Da, and then ultrafiltered with an ultrafiltration membrane with a molecular weight cutoff of 100 Da to obtain the ultrafiltrate. (4) The ultrafiltrate was freeze-dried under vacuum to obtain garlic powder; (5) Add pullulan, galactose, and polycarboxylic acid to water, stir well, then add branched starch, chitosan and maltodextrin, stir well to obtain shell liquid; (6) Add garlic powder to the shell liquid, stir evenly with ultrasound, and freeze dry under vacuum to obtain core-shell structured garlic powder.
2. The method for preparing core-shell structured garlic powder according to claim 1, characterized in that, The mass ratio of garlic to water is 1:(2~10).
3. The method for preparing core-shell structured garlic powder according to claim 1, characterized in that, The mass ratio of the compound enzyme to the slurry is (0.008~0.02):1; and / or The complex enzyme comprises cellulase, neutral protease, and papain in a mass ratio of 1:(0.5~1.2):(0.5~0.8).
4. The method for preparing core-shell structured garlic powder according to claim 1, characterized in that, The enzymatic hydrolysis is performed at a temperature of 45-50℃ for 0.5-2 hours.
5. The method for preparing core-shell structured garlic powder according to claim 1, characterized in that, The vacuum freeze drying in step (4) specifically involves: first drying at a vacuum of 28~32 Pa and a temperature of -10~-5℃ for 0.5~2 hours; then drying at a vacuum of 60~65 Pa and a temperature of -10~-5℃ for 1~3 hours; then drying at a vacuum of 12~16 Pa and a temperature of -2~2℃ for 0.5~1 hours; and finally drying at a vacuum of 8~12 Pa and a temperature of 18~22℃ for 1~3 hours.
6. The method for preparing core-shell structured garlic powder according to claim 1, characterized in that, The mass ratio of pullulan, galactose, polycarboxylic acid, water, amylopectin, chitosan, and hydroxypropyl-β-cyclodextrin is (2~3):1:(0.08~0.12):(15~30):(0.1~0.2):(0.8~1.5).
7. The method for preparing core-shell structured garlic powder according to claim 1, characterized in that, The mass ratio of garlic powder to shell liquid is 1:(3~4).
8. The method for preparing core-shell structured garlic powder according to claim 1, characterized in that, The vacuum freeze drying in step (6) specifically involves: first drying at a vacuum of 32~38 Pa and a temperature of -16~-14℃ for 0.5~2 hours; then drying at a vacuum of 45~50 Pa and a temperature of -8~-5℃ for 1~2 hours; then drying at a vacuum of 12~16 Pa and a temperature of -2~2℃ for 0.5~2 hours; and finally drying at a vacuum of 8~12 Pa and a temperature of 22~26℃ for 1~3 hours.
9. A core-shell structured garlic powder, characterized in that, It is prepared by any one of the preparation methods described in claims 1 to 8.
10. The use of the core-shell structured garlic powder of claim 9 in the preparation of a medicament for treating non-alcoholic steatohepatitis.