Use of a composition of cyanidin-3-o-glucoside in reducing hepatotoxicity caused by dietary heterocyclic amines

Abstract

The application discloses application of a composition of cyanidin-3-O-glucoside (C3G) in reducing liver toxicity caused by dietary heterocyclic amine IQ, and belongs to the field of food safety and toxicology. The composition is composed of C3G, rosemary extract, sesamin, curcumin, tea polyphenol and vitamin E in a specific weight ratio. A special process combining step-by-step premixing and equal-increment main mixing is adopted for preparation, so that uniform dispersion and interaction of the components are ensured. The composition can significantly improve liver function damage, inflammatory response and oxidative stress induced by IQ through multi-target synergistic effect. The application provides a new scheme for developing medicines, health foods or functional foods for relieving liver toxicity of IQ and other dietary carcinogens.

Description

Technical Field

[0001] This invention relates to the fields of food safety and toxicology, specifically to a novel use of a cyanidin-3-O-glucoside (C3G) composition, and particularly to the use of this composition in the preparation of products for reducing or preventing hepatotoxicity induced by dietary heterocyclic amine 2-amino-3-methylimidazo[4,5-f]quinoline (IQ). Background Technology

[0002] Heterocyclic amines are a class of compounds with strong mutagenic and carcinogenic properties, produced during high-temperature cooking (such as frying, grilling, and baking) of protein-rich foods. Among them, 2-amino-3-methylimidazo[4,5-f]quinoline (IQ) is one of the most toxic, classified as a Group 2A possible human carcinogen by the International Agency for Research on Cancer (IARC), and its toxicity and metabolic characteristics have attracted much attention. After ingestion, IQ is mainly metabolized and activated in the liver through the cytochrome P450 enzyme system, converting into highly reactive N-hydroxy derivatives, which can then trigger the formation of DNA adducts, oxidative stress, release of inflammatory mediators, and activation of multiple pathways of apoptosis, ultimately leading to hepatocyte damage and even carcinogenesis.

[0003] Currently, control strategies for the hazards of heterocyclic amines in food mainly focus on pre-processing interventions, such as improving heating methods and adding antioxidants or natural extracts to inhibit their formation. However, these methods cannot address the endogenous toxicity issues caused by heterocyclic amines that have already been ingested. For heterocyclic amines such as IQ that have already entered the body, there are currently no safe and effective intervention methods. Therefore, developing functional ingredients that can directly alleviate their in vivo toxicity is of great significance.

[0004] Anthocyanins are a class of water-soluble flavonoids widely found in dark-colored fruits and vegetables (such as blueberries, purple sweet potatoes, and black rice), with C3G being the most common. Existing studies generally report that anthocyanins possess antioxidant, anti-inflammatory, and anti-apoptotic biological activities, and show potential for protection against chemically induced liver injury. However, there are currently no reports on the application of C3G compositions in the preparation of products that alleviate IQ-induced liver injury. Therefore, developing a natural active ingredient or composition that can be applied in the post-intake phase of IQ to reduce its metabolic toxicity not only has significant theoretical value but also provides a new technical approach for developing functional foods targeting liver injury caused by dietary carcinogens. Summary of the Invention

[0005] The purpose of this invention is to address the shortcomings of existing technologies by revealing for the first time the novel use of anthocyanin-containing compositions, particularly anthocyanins represented by C3G, in reducing hepatotoxicity caused by dietary heterocyclic amines, thus providing a new solution for the development of related hepatoprotective products.

[0006] The technical solution of this invention is to provide an application of a C3G composition in the preparation of products for reducing hepatotoxicity caused by dietary heterocyclic amine IQ. The core of this invention is to provide a compound system with C3G as the main component and multiple active ingredients working synergistically to alleviate IQ-induced liver damage through multiple targets. The active ingredients of the composition include C3G, rosemary extract, sesamin, curcumin, tea polyphenols, and vitamin E; the product includes a drug, health food, or functional food; the application is specifically reflected in the specific composition, preparation method, and its role in synergistically reducing hepatotoxicity through multiple pathways.

[0007] In a first aspect, the present invention discloses a C3G composition, wherein the composition comprises, by weight parts, the following active ingredients: C3G (purity ≥80%) 20–40 parts; Rosemary extract (≥20% based on rosmarinic acid) 5–15 parts; Sesamin (purity ≥40%) 5–10 parts; Curcumin (purity ≥90%) 1–5 parts; Tea polyphenols (≥50% as EGCG) 3–10 parts; Vitamin E (d-α-tocopherol) 1–5 parts.

[0008] Preferably, the active ingredients in the C3G composition include, by weight: C3G (purity ≥80%) 25–35 parts; 8–12 parts of rosemary extract (≥20% based on rosmarinic acid); Sesamin (purity ≥40%) 6–8 parts; Curcumin (purity ≥90%) 2–4 parts; Tea polyphenols (≥50% as EGCG) 5–8 parts; Vitamin E (d-α-tocopherol) 2–4 parts.

[0009] More preferably, the active ingredients in the composition include, by weight: 30 portions of C3G (purity ≥80%); 10 parts of rosemary extract (≥20% based on rosmarinic acid); Sesamin (purity ≥40%) 5 parts; Curcumin (purity ≥90%) 3 parts; Six portions of tea polyphenols (≥50% as EGCG); Vitamin E (d-α-tocopherol) 2 parts.

[0010] In a second aspect, the present invention provides a method for preparing the C3G composition as described in the first aspect, the method comprising the following steps: 1. Raw material pretreatment (1) Pretreatment of vitamin E: Take the prescribed amount of oily vitamin E. If the viscosity at 25°C is >50 mPa·s, heat it in a water bath at 30-40°C for 5-15 minutes, stir until the viscosity drops to 20-50 mPa·s, and cool it to room temperature for later use; if the viscosity is ≤50 mPa·s, use it directly. (2) Preparation of vitamin E adsorbent: Sesamin was used as a carrier and added to a premixing device to be mixed and adsorbed with vitamin E. The mixture was then ground or stirred until a uniform loose powder without visible oil droplets was formed. (3) Pretreatment of powder components: The C3G, rosemary extract, curcumin and tea polyphenols in the specified weight parts are passed through an 80-120 mesh standard sieve to remove lumps, so as to ensure that the basic particle size of each component is similar and reduce the separation phenomenon caused by particle size difference.

[0011] 2. Stepwise premixing (1) Preparation of the first premix: The curcumin and tea polyphenols in the specified weight parts are put into a mixer and mixed at a speed of 20-40 rpm for 15-25 min to obtain the first premix; (2) Preparation of the second premix: The vitamin E adsorbent and the amount of rosemary extract by weight are put into another mixer or the same mixer after cleaning, and mixed at a speed of 20-40 rpm for 10-20 min to obtain the second premix.

[0012] 3. Principal mixture with equal increasing amounts (1) Put all the main C3G materials into the mixer; (2) Using the equal incremental method, first add C3G main material with a volume approximately equal to that of the second premix and mix it with the second premix. Then, put all of this mixture into the main mixer and mix it with the material in the machine for 10-15 minutes. (3) Using the equal incremental method again, take out the current total mixture with a volume approximately equal to that of the first premix, mix it initially with the first premix, and then put it all into the main mixer; (4) Finally, mix at a speed of 20-40 rpm for 30-50 min until the mixture has a uniform color.

[0013] 4. Post-mixing treatment The mixing process is carried out in an environment with relative humidity ≤45% and temperature ≤25℃. After mixing, the material is discharged and immediately sealed in aluminum foil bags, and desiccant and light-proof outer packaging are provided to maximize the protection of the active ingredients from the effects of moisture, oxygen and light.

[0014] Thirdly, the C3G composition reduces hepatotoxicity through multiple pathways: 1. Improves liver function and reduces serum transaminase (AST, ALT) levels; 2. It inhibits liver inflammation and reduces the levels of inflammatory factors (TNF-α, IL-1β, IL-6) in serum; 4. Reduces liver oxidative stress, increases the activity of glutathione peroxidase (GSH-Px) and superoxide dismutase (SOD) in liver tissue, and reduces malondialdehyde (MDA) levels. The beneficial effects of this invention are:

[0015] 1. Synergistic Effect of Multiple Targets: This invention is not a simple combination of multiple ingredients, but a scientific formulation based on their different mechanisms of action. Each ingredient complements and synergistically enhances the effects of anti-oxidation and anti-inflammation, resulting in significantly better effects than a single ingredient.

[0016] 2. Pioneering Application: This is the first time that the combination of the above six natural active ingredients has been applied to the specific field of antagonizing IQ hepatotoxicity, opening up new uses for the composition and providing a brand-new solution to address the health risks following dietary IQ exposure.

[0017] 3. High safety: All active ingredients are of natural origin or recognized safe food additives / nutrients, with high safety and high consumer acceptance. It is very suitable for developing health products or functional foods for consumption during the growth period and is suitable for people who frequently consume barbecued and fried foods.

[0018] 4. Diverse application forms: This composition can be flexibly applied to various forms of health foods or functional foods such as capsules, tablets, and powders, and has broad market application prospects. Detailed Implementation

[0019] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the embodiments described are merely illustrative of the present invention and should not be construed as limiting the invention. Example 1

[0020] This embodiment provides a C3G composition, the active ingredients of which, by weight, are as follows: 30 parts of C3G (purity ≥80%), 10 parts of rosemary extract (≥20% based on rosmarinic acid), 5 parts of sesamin (purity ≥40%), 3 parts of curcumin (purity ≥90%), 6 parts of tea polyphenols (≥50% based on EGCG), and 2 parts of vitamin E (d-α-tocopherol).

[0021] The method for preparing the composition includes the following steps: 1. Raw material pretreatment (1) Vitamin E pretreatment: Take the amount of vitamin E in the formula, heat it in a water bath at 35°C for 10 min, stir until the viscosity is about 30 mPa·s, and cool it to room temperature for later use; (2) Preparation of vitamin E adsorbent: Take 5 parts of sesamin as carrier and mix it with 2 parts of pretreated vitamin E in a low-speed shear mixer for 15 min until a uniform, oil-free loose powder is formed.

[0022] (3) Pretreatment of powder components: 30 parts of C3G, 10 parts of rosemary extract, 3 parts of curcumin and 6 parts of tea polyphenols were passed through a 100-mesh sieve to break up the clumps and set aside.

[0023] 2. Stepwise premixing (1) Preparation of the first premix: 3 parts curcumin and 6 parts tea polyphenols were put into a three-dimensional motion mixer and mixed at 30 rpm for 20 min; (2) Preparation of the second premix: The vitamin E adsorbent and 10 parts of rosemary extract were put into another mixer and mixed at 30 rpm for 15 min.

[0024] 3. Principal mixture with equal increasing amounts All C3G was put into the three-dimensional motion mixer. First, the second premix was added in using the equal-increment method and mixed for 12 minutes. Then, the first premix was added in the same way. Finally, the mixture was mixed at 30 rpm for 35 minutes until the mixture was uniform in color.

[0025] 4. Post-mixing treatment Mixing was carried out at 25℃ and humidity ≤45%. Immediately after discharge, the material was packed into aluminum-plastic composite bags, 3 g of montmorillonite desiccant was added, and the bags were vacuum-sealed (vacuum degree 0.095 MPa). The bags were then stored in brown light-proof bags. Example 2

[0026] This embodiment provides a C3G composition, the active ingredients of which, by weight, are as follows: 35 parts of C3G (purity ≥80%), 12 parts of rosemary extract (≥20% based on rosmarinic acid), 8 parts of sesamin (purity ≥40%), 4 parts of curcumin (purity ≥90%), 8 parts of tea polyphenols (≥50% based on EGCG), and 4 parts of vitamin E (d-α-tocopherol).

[0027] The method for preparing the composition includes the following steps: 1. Raw material pretreatment (1) Vitamin E pretreatment: Viscosity 55 mPa·s (25℃), place in a 35℃ water bath and heat for 10 min, stir until the viscosity drops to 45 mPa·s, cool and set aside; (2) Preparation of vitamin E adsorbent: Take 8 parts of sesamin and 2 parts of vitamin E and add them to a low-speed shear mixer and mix for 12 min to obtain a loose adsorbent without oil spots; (3) Pretreatment of powder components: 35 parts of C3G, 12 parts of rosemary extract, 4 parts of curcumin and 8 parts of tea polyphenols were passed through a 100-mesh sieve to remove lumps.

[0028] 2. Stepwise premixing (1) Preparation of the first premix: 4 parts curcumin and 8 parts tea polyphenols were put into a double paddle mixer and mixed at 30 rpm for 20 min to obtain the first premix; (2) Preparation of the second premix: After cleaning the mixer, add vitamin E adsorbent and 12 parts of rosemary extract, mix at 30 rpm for 15 min to obtain the second premix.

[0029] 3. Principal mixture with equal increasing amounts Using 35 portions of C3G as the main component, the second and first premixes were added in equal increments and mixed at 25 rpm for 30 min.

[0030] 4. Post-mixing treatment The ambient temperature was 23°C and the humidity was 42%, and the rest was the same as in Example 1. Example 3

[0031] This embodiment provides a C3G composition, the active ingredients of which, by weight, are as follows: 20 parts of C3G (purity ≥80%), 5 parts of rosemary extract (≥20% based on rosmarinic acid), 5 parts of sesamin (purity ≥40%), 1 part of curcumin (purity ≥90%), 3 parts of tea polyphenols (≥50% based on EGCG), and 1 part of vitamin E (d-α-tocopherol).

[0032] The method for preparing the composition includes the following steps: 1. Raw material pretreatment The oily vitamin E has a viscosity of 40 mPa·s (25℃) and can be used directly; take 2 parts of sesamin to adsorb 1 part of vitamin E, and pass the remaining powder through an 80-mesh sieve.

[0033] 2. Stepwise premixing (1) Preparation of the first premix: The remaining 3 parts of sesamin, 1 part of curcumin and 3 parts of tea polyphenols were put into a double paddle mixer and mixed at 25 rpm for 10 min to obtain the first premix; (2) Preparation of the second premix: After cleaning the mixer, add vitamin E adsorbent and 5 parts of rosemary extract, mix at 25 rpm for 10 min to obtain the second premix.

[0034] 3. Principal mixture with equal increasing amounts Using 20 portions of C3G as the main component, the second and first premixes were added in equal increments and mixed at 25 rpm for 30 min.

[0035] 4. Post-mixing treatment The ambient temperature was 22°C and the humidity was 40%, and the rest was the same as in Example 1. Comparative Example 1

[0036] This comparative example illustrates the central role of C3G in this composition. Except for the absence of C3G, the other components and their amounts are identical to those in Example 1. Due to the lack of a bulk material, all other components were mixed directly in a single step using conventional methods during preparation. Comparative Example 2

[0037] This comparative example illustrates the synergistic effect of the compound system. The formulation contains 30 parts C3G (purity ≥80%), 3 parts curcumin (purity ≥90%), 6 parts tea polyphenols (≥50% as EGCG), and 2 parts vitamin E (d-α-tocopherol). Except for the absence of rosemary extract and sesamin, the other components and amounts are exactly the same as in Example 1. During preparation, vitamin E is adsorbed with an equal amount of microcrystalline cellulose, and the remaining processes are the same as in Example 1. Comparative Example 3

[0038] This comparative example illustrates the synergistic effect of the C3G composition of the present invention. The formulation uses only C3G (purity ≥80%) as the sole active ingredient. Comparative Example 4

[0039] This comparative example illustrates the necessity of the special mixing process of the present invention. The formulation is exactly the same as in Example 1. During preparation, all solid powder components (including sesamin for adsorbing vitamin E) are added to the mixer at once and mixed at 30 rpm for 60 min. Test case

[0040] Animal experiments were conducted to verify the protective effect of the C3G composition against IQ-induced hepatotoxicity.

[0041] 1. Animal grouping and treatment SPF-grade ICR male mice (weighing 18–22 g) were randomly divided into 9 groups of 10 mice each, according to their body weight: control group, model group, Example 1–3 groups, and Comparative Example 1–4 groups. All animals were treated continuously for 28 days, and various indicators were measured after the intervention.

[0042] The processing for each group is as follows: Control group: Corn oil administered by gavage; Model group: administered corn oil solution containing 20 mg / kg IQ by gavage; Examples 1–3: The corresponding combination of the example was administered by gavage (100 mg / kg), and simultaneously 20 mg / kg IQ was administered by gavage. Comparative Examples 1, 2, and 4: The corresponding comparative example composition (100 mg / kg) was administered by gavage, and 20 mg / kg IQ was administered by gavage simultaneously. Comparative group 3: 100 mg / kg C3G was administered by gavage, and 20 mg / kg IQ was administered by gavage simultaneously.

[0043] All IQ-containing gavage solutions use corn oil as a solvent.

[0044] 2. Detection Method (1) Liver function index detection: Blood was collected from the orbital bone of mice after the experiment, and serum was separated by centrifugation. The activities of AST (aspartate aminotransferase) and ALT (alanine aminotransferase) were detected using a kit (Nanjing Jiancheng Bioengineering Institute). (2) Detection of inflammatory factors: The levels of TNF-α (tumor necrosis factor-α), IL-1β (interleukin-1β) and IL-6 (interleukin-6) in serum were detected using an ELISA kit (Hangzhou Lianke Biotechnology Co., Ltd.); (3) Detection of oxidative stress indicators: Mouse liver tissue was taken, homogenized, and GSH-Px (glutathione peroxidase) and SOD (superoxide dismutase) activities and MDA (malondialdehyde) levels were detected using a kit (Nanjing Jiancheng Bioengineering Institute).

[0045] 3. Test Results This experiment evaluated the protective effect of different C3G compositions against IQ-induced hepatotoxicity in mice by detecting serum liver function and inflammatory factor indicators. The results are shown in Table 1.

[0046] Table 1 Comparison 15.84±2.06 3.16±0.47 93.60±0.32 76.79±5.52 44.35±5.90 Model 19.60±1.32 5.46±0.67 101.17±1.17 90.26±7.93 66.62±6.07 Example 1 14.52±1.65 2.85±0.42 92.55±1.25 75.12±6.32 42.18±4.87 Example 2 15.23±1.88 3.02±0.58 93.20±1.41 76.00±5.94 44.52±5.23 Example 3 15.87±1.92 3.15±0.51 94.05±1.63 75.89±6.81 46.75±5.60 Comparative Example 1 18.52±1.75 4.08±0.62 99.85±1.88 88.45±7.25 62.34±6.45 Comparative Example 2 16.02±1.83 3.27±0.66 95.90±1.70 80.15±6.50 51.20±5.88 Comparative Example 3 16.24±3.22 3.44±1.29 95.83±2.09 79.46±10.56 52.19±5.63 Comparative Example 4 16.25±1.78 3.42±0.59 96.08±1.95 80.40±7.02 52.80±6.10 AST and ALT activities are key indicators for evaluating the degree of hepatocyte damage. The model group showed the highest activities of these two indicators, indicating significant liver damage in mice after IQ administration. All intervention groups exhibited varying degrees of protective effects, with the example groups showing the best results: Example 1 had the lowest AST and ALT levels, followed by Examples 2 and 3, indicating that all three C3G compositions have strong hepatoprotective effects. The comparative groups were generally less effective than the example groups, especially Comparative Example 1, which lacked the core component C3G and thus had the weakest protective effect. Comparative Example 2 (lacking synergistic components) and Comparative Example 3 (single C3G) showed some protective effects, but significantly less than Examples 1 and 2, demonstrating the crucial role of compound formulation in achieving synergistic effects.

[0047] TNF-α, IL-1β, and IL-6 are core pro-inflammatory cytokines, and elevated levels reflect activation of the inflammatory response. Compared with the control group, the levels of these three inflammatory factors were all increased in the model group, indicating that IQ induced a strong inflammatory response. The example groups showed excellent anti-inflammatory effects: Example 1 was particularly outstanding, with the levels of the three inflammatory factors being much lower than those in the model group and comparable to those in the control group, indicating that it almost completely inhibited IQ-induced inflammation; Examples 2 and 3 also showed strong anti-inflammatory effects, with each indicator similar to the control group and Example 1, and the effects were significantly better than all comparative groups. The anti-inflammatory effects of the comparative groups were all inferior to those of the example groups, with Comparative Example 1 (lacking the core component C3G) showing the worst effect and weak anti-inflammatory effect; Comparative Examples 2 and 3 were significantly better than the model group and Comparative Example 1, but still lagged behind the example groups, indicating that while single C3G or incomplete formulations have certain anti-inflammatory effects, they are far inferior to complete C3G compositions.

[0048] The comparison of the effects of Comparative Example 4 (mixed simultaneously) and Example 1 (mixed sequentially) shows that the preparation process has a decisive influence on the efficacy of the composition. The liver function improvement and anti-inflammatory effects of Comparative Example 4 were significantly lower than those of Example 1, indicating a weakened protective effect. The stepwise mixing process can more effectively ensure that the components are uniformly dispersed and fully interact, while the simple "one-pot mixing" method cannot achieve the same synergistic effect, resulting in a decrease in the efficacy of the final product.

[0049] In summary, the C3G compositions (Examples 1-3) significantly improved IQ-induced hepatocellular damage and inflammatory responses, and promoted liver function recovery, with effects significantly superior to single-component formulations (Comparative Example 3) or incomplete formulations (Comparative Examples 1-2). The formulation and process represented by Example 1 exhibited the best protective efficacy, strongly demonstrating the necessity and superiority of the compound formulation and special process in this invention. The absence of any component or process (Comparative Examples 1-4) significantly reduced the protective effect against IQ-induced hepatotoxicity in mice.

[0050] Furthermore, this experiment evaluated the protective effect of different C3G compositions against IQ-induced hepatotoxicity by detecting antioxidant indices in mouse liver tissue. The specific results are shown in Table 2.

[0051] Table 2 Comparison 371.79±77.29 166.22±22.00 0.79±0.07 Model 226.65±29.67 148.85±13.30 1.52±0.30 Example 1 398.25±70.15 182.50±25.40 0.72±0.09 Example 2 375.44±68.33 178.63±20.18 0.85±0.11 Example 3 350.12±60.27 175.88±18.55 0.90±0.15 Comparative Example 1 310.18±35.20 162.30±15.62 1.12±0.25 Comparative Example 2 340.50±50.41 169.75±17.83 1.09±0.20 Comparative Example 3 335.80±45.16 168.90±19.50 1.02±0.18 Comparative Example 4 339.05±43.63 171.70±22.07 1.05±0.24 As shown in Table 2, the GSH-Px and SOD activities in the model group were the lowest, indicating that IQ exposure significantly inhibited the antioxidant enzyme system in the mouse liver, leading to a decrease in the body's ability to scavenge free radicals. The two enzyme activities in Example 1 group were the highest, even higher than the normal control group, indicating that the C3G composition not only completely resisted oxidative damage caused by IQ but also significantly enhanced the body's antioxidant defense capabilities. The enzyme activities in Example 2 and 3 groups also remained at a high level, comparable to or slightly higher than the normal control group, showing good antioxidant effects. The enzyme activity in Comparative Example 1 (lacking the core component C3G) was significantly lower than that in the example groups, indicating that its effect on enhancing antioxidant capacity was limited. The enzyme activities in Comparative Examples 2, 3, and 4 were between those of the model group and the example groups, indicating that they had some antioxidant effects, but the effect was far less than that of the complete C3G composition (Examples 1-3).

[0052] The model group had the highest MDA content, indicating that IQ successfully induced severe lipid peroxidation damage in hepatocytes. The Example 1 group had the lowest MDA content, indicating that it effectively inhibited lipid peroxidation and had the strongest protective effect on hepatocytes. The MDA content in Example 2 and 3 groups also remained at low levels, showing a strong protective effect against oxidative damage. The MDA content in Comparative Example 1 group was much higher than that in the Example groups, and although slightly lower than that in the model group, it was still at a high level, indicating that its effect in alleviating oxidative damage was poor. The MDA content in Comparative Examples 2, 3, and 4 was significantly lower than that in the model group, but higher than that in the Example groups, indicating that it could only partially alleviate IQ-induced oxidative damage.

[0053] The absence of any core ingredient (Comparative Example 1), incomplete formulation (Comparative Examples 2 and 3), or different preparation processes (Comparative Example 4, i.e., all components are mixed simultaneously) all lead to a significant decrease in its antioxidant efficacy. This indicates that the present invention relies not only on the integrity and synergy of the compound formulation, but also on an optimized stepwise mixing process; the combined effect of these two factors is key to ensuring that the C3G composition achieves the best liver protection effect. Application Example 1

[0054] This application example provides a method for preparing the C3G composition in powder form according to the present invention.

[0055] Following the method described in Example 1, all active ingredients were mixed evenly to obtain the C3G composite powder. The composite powder was then added to a mixer with erythritol (43.5 parts) and mixed for 15 minutes. Silica (0.5 parts) and fragrance (appropriate amount) were added, and the mixture was mixed for another 5 minutes. The entire process was carried out under controlled temperature and humidity conditions. The mixed powder was immediately sieved through a 100-mesh sieve and then automatically packaged into aluminum foil bags, each containing 1.0 g of powder product. Application Example 2

[0056] This application example provides a method for preparing the C3G composition of the present invention in hard capsule form.

[0057] Following the method described in Example 1, all active ingredients were mixed evenly to obtain a C3G composite powder. Microcrystalline cellulose (94 parts), hydroxypropyl methylcellulose (5 parts), and crospovidone (3 parts) were added to the composite powder and mixed for 30 min. Subsequently, micronized silica gel (0.5 parts) and magnesium stearate (1.5 parts) were added, and the mixture was further mixed. The mixture was then filled into No. 1 empty capsules using a fully automated capsule filling machine, with each capsule containing 100 mg of total active ingredients. Application Example 3

[0058] This application example provides a method for preparing the C3G composition of the present invention in tablet form.

[0059] Following the method described in Example 1, all active ingredients were mixed evenly to obtain a C3G composite powder. Lactose (120 parts) and microcrystalline cellulose (60 parts) were added to the composite powder and mixed for 15 min. A 5% povidone K30 ethanol solution (3 parts) was added to form a soft mass, which was granulated through a 20-mesh sieve, dried at 50°C, and then granulated through an 18-mesh sieve. Subsequently, cross-linked povidone (6 parts) and microcrystalline silica (0.5 parts) were added and mixed for 10 min. Finally, magnesium stearate (1.5 parts) was added and mixed for 5 min, and the mixture was compressed into tablets weighing 500 mg each. The tablets were then coated with a film-coating solution composed of hydroxypropyl methylcellulose (5 parts) and titanium dioxide (0.5 parts) to obtain the finished product, in which the total active ingredient content is approximately 100 mg / tablet.

[0060] The applicant declares that the technical solution and effects of the present invention have been described in detail through the above embodiments, but the present invention is not limited to the specific embodiments described. That is, it does not mean that the present invention must rely on the specific methods or formulas described above to be implemented. Those skilled in the art should understand that, within the technical concept and core principles of the present invention, any form of improvement and modification, such as adjustments to the method steps, equivalent substitutions of each raw material component, additions, subtractions or substitutions of auxiliary components, and reasonable changes to specific process parameters, falls within the protection scope and disclosure scope of the present invention.

Claims

1. The use of a cyanidin-3-O-glucoside (C3G) composition in the preparation of a product for reducing or preventing hepatotoxicity caused by the dietary heterocyclic amine 2-amino-3-methylimidazo[4,5-f]quinoline (IQ), characterized in that, The active ingredients of the composition consist of the following components in parts by weight: 20–40 parts of C3G (purity ≥80%); 5–15 parts of rosemary extract (≥20% based on rosmarinic acid); 5–10 parts of sesamin (purity ≥40%); 1–5 parts of curcumin (purity ≥90%); 3–10 parts of tea polyphenols (≥50% based on EGCG); and 1–5 parts of vitamin E (d-α-tocopherol).

2. The application according to claim 1, characterized in that, The active ingredients of the composition consist of the following components in parts by weight: C3G 25–35 parts; rosemary extract 8–12 parts; sesamin 6–8 parts; curcumin 2–4 parts; tea polyphenols 5–8 parts; vitamin E 2–4 parts.

3. The application according to claim 1, characterized in that, The active ingredients of the composition consist of the following components in parts by weight: 30 parts C3G; 10 parts rosemary extract; 5 parts sesamin; 3 parts curcumin; 6 parts tea polyphenols; and 2 parts vitamin E.

4. A method for preparing the C3G composition according to any one of claims 1-3, characterized in that, The method includes the following steps: (1) Raw material pretreatment: including adjusting the oily vitamin E to a suitable viscosity and using some sesamin as a carrier for adsorption to form vitamin E adsorbate; and sieving C3G, rosemary extract, curcumin and tea polyphenols to break up clumps. (2) Stepwise premixing: Curcumin and tea polyphenols are premixed to obtain a first premix; the vitamin E adsorbent is premixed with rosemary extract to obtain a second premix; (3) Equal-incremental main mixing: Using all C3G as the main material, the second premix and the first premix are added to the mixture in sequence using the equal-incremental method until the mixture has a uniform color; (4) Post-mixing treatment: The entire mixing process is carried out in an environment with controlled temperature and humidity. After mixing, the mixture is sealed and packaged in a light-proof container.

5. The method according to claim 4, characterized in that, If the viscosity of the oily vitamin E described in step (1) is greater than 50 mPa·s at 25°C, it should be heated in a water bath at 30-40°C for 5-15 minutes, stirred until the viscosity drops to 20-50 mPa·s, and then cooled to room temperature before the adsorption operation is carried out.

6. The method according to claim 4, characterized in that, In step (2), the first premix and the second premix are both carried out at a speed of 20-40 rpm, and the mixing time is 15-25 min and 10-20 min, respectively.

7. The method according to claim 4, characterized in that, The main mixing in step (3) is carried out at a speed of 20-40 rpm for a mixing time of 30-50 min; and the relative humidity of the environment during the entire mixing process is ≤45% and the temperature is ≤25℃.

8. The use of the composition according to claims 1-3 in the preparation of medicaments, health foods or functional foods for preventing or alleviating IQ-induced liver injury.

9. A product for preventing or alleviating IQ-induced liver damage, characterized in that, It comprises the C3G composition as described in any one of claims 1-3 and pharmaceutically or food-grade excipients.

10. The product according to claim 9, characterized in that, The product is available in powder, capsule, or tablet form.