Multi-target synergistic cardiovascular protection beverage and preparation method thereof

Abstract

The invention provides a multi-target synergistic cardiovascular protection beverage which comprises functional components and auxiliary components, and the functional components comprise nattokinase, hirudin, squalene, EPA, DHA and phytosterol; the preparation method of the multi-target synergistic cardiovascular protection beverage comprises the following steps: S1, firstly, washing the functional components with deionized water respectively to remove surface impurities, dust and microorganisms for subsequent processing; compared with the prior art, the raw material drying device has the following beneficial effects that in the drying process, flowing of hot air can take away moisture on the surfaces of raw materials, the humidity gradient is formed, the drying process is more uniform, and the phenomenon that the raw materials are locally overheated or dried unevenly is avoided; the method disclosed by the invention has the following beneficial effects that the raw materials are washed by deionized water and are dried by vacuum or low-temperature hot air, so that impurities and microorganisms can be thoroughly removed, and the loss of heat-sensitive functional components can be reduced.

Description

Technical Field

[0001] This invention belongs to the field of cardiovascular protective beverage technology, and relates to a multi-target synergistic cardiovascular protective beverage and its preparation method. Background Technology

[0002] Existing cardiovascular protective beverages suffer from core drawbacks in cardiovascular protection: they target only a single efficacy point, focusing on the action of a single component and lacking a multi-component synergistic mechanism. This results in limited core efficacy in areas such as thrombolysis, lipid regulation, and vascular endothelial protection, and insufficient comprehensive protection of the myocardium. Furthermore, some products suffer from issues such as loss of active ingredient activity, low purity, and unpleasant taste. These shortcomings arise from two main causes: firstly, the formulation design does not consider the multi-dimensional needs of thrombolysis, lipid regulation, and vascular endothelial repair, making it difficult for a single component to cover the entire cardiovascular protection chain; secondly, the pretreatment, extraction, and processing of raw materials are not precise enough. Traditional cleaning methods cannot thoroughly remove impurities and microorganisms, high-temperature drying or extraction can damage heat-sensitive components, and ordinary mixing processes lead to uneven component dispersion, affecting synergistic effects. Conventional solutions include increasing the dosage of single components, using traditional organic solvent extraction, adding large amounts of sweeteners to mask off-flavors, and simplifying processing procedures to reduce costs. However, these methods have obvious drawbacks. Increasing the dosage of a single ingredient can easily cause side effects, and traditional organic solvent extraction can leave harmful substances, affecting product safety. Therefore, there is an urgent need for a multi-target synergistic cardiovascular protection beverage and its preparation method to solve the above problems. Summary of the Invention

[0003] To address the shortcomings of existing technologies, the present invention aims to provide a multi-target synergistic cardiovascular protective beverage and its preparation method, thereby resolving the problems mentioned in the background section.

[0004] This invention is achieved through the following technical solution: a multi-target synergistic cardiovascular protective beverage, comprising: active ingredients and auxiliary ingredients; The active ingredients include nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2, and phytosterols; The product composition includes nattokinase (0.015% of total mass), hirudin (0.003%), squalene (0.43%), EPA (0.3%), DHA (0.2%), red yeast rice extract (0.8%), coenzyme Q10 (0.05%), vitamin K2 (0.001%), and phytosterols (1.2%). Nattokinase directly breaks down fibrin thrombi, hirudin strongly inhibits thrombin activity, blocking the thrombus formation chain. Combined with the antiplatelet aggregation effects of EPA and DHA, it reduces the risk of venous thrombosis and atherosclerotic thrombosis. Red yeast rice extract also inhibits... It inhibits cholesterol synthesis, phytosterols compete for cholesterol absorption in the intestines, and EPA and DHA regulate triglyceride metabolism. The three work synergistically to lower total cholesterol and LDL cholesterol while raising HDL cholesterol. Squalene enhances blood vessel wall elasticity, reduces oxidative damage, and slows down the process of arteriosclerosis. At the same time, nattokinase has extremely strong fibrinolytic activity. At a dose of 0.015%, drinking 200-300g of beverage daily can provide 30-45mg of nattokinase, which meets the standard for effective daily intake for adults. At the same time, it avoids coagulation disorders caused by excessive doses, making it suitable for long-term consumption. Nattokinase and hirudin are the core components. The fibrinolytic effect of nattokinase and the anticoagulant effect of hirudin complement each other, ensuring thrombolytic effect while avoiding the risks of excessive intake of a single component. The auxiliary ingredients include fructooligosaccharides, maltitol, xanthan gum, CMC-Na, citric acid, sodium citrate, food flavoring, and purified water; The total mass of the finished product is as follows: fructooligosaccharides account for 3.00%, maltitol accounts for 5%, xanthan gum accounts for 0.15%, CMC-Na accounts for 0.1%, citric acid accounts for 0.2%, sodium citrate accounts for 0.08%, edible flavoring accounts for 0.027%, and purified water accounts for 88.594%.

[0005] The beneficial effects of this invention after adopting the above technical solution are as follows: Nattokinase directly decomposes fibrin thrombi; hirudin strongly inhibits thrombin activity, blocking the thrombus formation chain; combined with the antiplatelet aggregation effects of EPA and DHA, it reduces the risk of venous thrombosis and atherosclerotic thrombosis; red yeast rice extract inhibits cholesterol synthesis; phytosterols compete for intestinal cholesterol absorption; EPA and DHA regulate triglyceride metabolism; and the three work synergistically to reduce total cholesterol and low-density lipoprotein while increasing high-density lipoprotein; squalene enhances vascular wall elasticity and reduces... It reduces oxidative damage and slows down the process of arteriosclerosis. At the same time, nattokinase has extremely strong fibrinolytic activity. At a dose of 0.015%, drinking 200-300g of beverage daily can provide 30-45mg of nattokinase, which meets the standard daily effective intake for adults. At the same time, it avoids coagulation disorders caused by excessive doses, making it suitable for long-term consumption. Nattokinase and hirudin are the core components. The fibrinolytic effect of nattokinase and the anticoagulant effect of hirudin complement each other, ensuring thrombolytic effect while avoiding the risks of excessive intake of a single component.

[0006] A method for preparing a multi-target synergistic cardiovascular protective beverage includes the following steps: S1: First, rinse the active ingredients separately with deionized water to remove surface impurities, dust and microorganisms, and then dry the cleaned raw materials to remove moisture for subsequent processing; S2: According to the proportions in the formula framework, use a high-precision electronic balance to accurately weigh nattokinase extract, hirudin extract, squalene, EPA and DHA and other auxiliary ingredients. Add the weighed ingredients to a mixing tank with a stirring device. First, stir at low speed to initially disperse the ingredients evenly. After initial mixing, turn on a high-speed homogenizer to further refine the particles and fully mix the ingredients. S3: The homogenized mixture is then concentrated and transferred to the evaporation flask of a rotary evaporator for further concentration to increase the concentration of active ingredients. Based on the expected taste of the product, appropriate amounts of sweeteners and acidulants are added for taste adjustment. The content of various nutrients in the concentrated beverage is then tested. S4: The prepared beverage undergoes a UHT sterilization process, and PET plastic bottles or glass bottles are used as packaging containers. Then, the filling process is carried out in a sterile workshop. After filling, the caps or seals are applied immediately to ensure that the beverage is sealed.

[0007] As a preferred embodiment, nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2, and phytosterols are rinsed separately with deionized water for 3-5 minutes to remove surface impurities, dust, and microorganisms. The cleaned raw materials are then dried to remove moisture. The drying method is vacuum drying or low-temperature hot air drying. During the rinsing process, the water flow can be evenly contacted to all parts of the raw materials by gentle manual stirring or by using specialized cleaning equipment to ensure a comprehensive cleaning effect. During vacuum drying, the pressure is set at 0.05-0.1 MPa, the temperature at 40-50℃, and the drying time at 3-5 hours. In a vacuum environment, the boiling point of water decreases, allowing it to vaporize rapidly at a lower temperature. Due to the lower drying temperature, the loss of heat-sensitive components is effectively avoided, thus achieving rapid drying. In low-temperature hot air drying, hot air is used as the drying medium, and the moisture in the raw material evaporates through heat transfer. The temperature is controlled at 50-60℃, and the air velocity is 0.5-1.0 m / s. Drying continues until the moisture content of the raw material is below 5%. The raw materials are pulverized into fine powder using a pulverizer. During the drying process, the flow of hot air can remove moisture from the surface of the raw materials, forming a humidity gradient and accelerating the evaporation of moisture. By controlling the wind speed and temperature, the relevant staff can make the drying process more uniform, avoid local overheating or uneven drying of the raw materials, and improve the subsequent extraction efficiency. The particle size of nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2 and phytosterols after pulverization is 80-100 mesh, which facilitates the full extraction of effective ingredients.

[0008] In a preferred embodiment, the nattokinase is prepared by crushing natto. The crushed natto is then added to 10-15 times its volume of deionized water and extracted by stirring at 37-40°C for 4-6 hours. Since this temperature range is close to human body temperature, it is a relatively stable temperature range for nattokinase activity, which can maximize the release of nattokinase. The stirring speed is 100-150 r / min. After centrifugation at 3000-5000 r / min for 15-20 minutes, insoluble impurities are precipitated, resulting in a relatively clear supernatant. The supernatant contains nattokinase and some other impurities. The supernatant is then ultrafiltered through an ultrafiltration membrane with a molecular weight cutoff of 10-20 kDa. The ultrafiltration membrane separates the components in the solution according to molecular size. Only substances with a molecular weight smaller than the cutoff molecular weight can pass through the ultrafiltration membrane. The molecular weight of nattokinase is exactly within this range, thus achieving rapid and effective separation and obtaining an extract rich in nattokinase with a nattokinase activity ≥2000 IU / mg. Hirudin is prepared by placing leech powder in an extraction vessel with CO2 as the extractant. The extraction pressure is 20-30 MPa, the temperature is 40-50℃, and the extraction time is 2-3 hours. The separation vessel pressure is 5-10 MPa and the temperature is 30-40℃. Under supercritical conditions, CO2 has a density similar to that of a liquid and a diffusion coefficient similar to that of a gas, which can quickly penetrate into the interior of the leech powder, fully contact with the hirudin, and dissolve it. The higher extraction pressure and suitable temperature can enhance the solubility of CO2, improve the extraction efficiency of hirudin, and obtain a high-purity hirudin extract. The purity of the finished hirudin is ≥95%. At the same time, the supercritical carbon dioxide extraction technology avoids the residue problems caused by traditional organic solvent extraction, ensuring the purity and safety of hirudin, while also improving extraction efficiency and reducing extraction time and energy consumption. The EPA and DHA are extracted from fish oil and algal oil using a low-temperature enzymatic hydrolysis combined with molecular distillation. An appropriate amount of lipase is added to the fish oil or algal oil at a lipase-to-substrate mass ratio of 1:100-1:200. Enzymatic hydrolysis is performed at 40-45℃ and pH 7.0-8.0 for 3-5 hours. Under these conditions, the lipase specifically acts on the triglycerides in the fish oil or algal oil, breaking them down into fatty acids and glycerol, thus releasing EPA and DHA from the triglycerides. Suitable temperature and pH are key factors for lipase activity, ensuring the smooth progress of the enzymatic hydrolysis reaction. After centrifugation, the supernatant is subjected to molecular distillation. Distillation is carried out at 120-150℃ and 0.1-0.5Pa. Light fractions are collected to obtain a product rich in EPA and DHA. The red yeast rice extract is selected with a lovastatin content of ≥1.5% and a coenzyme Q10 purity of ≥98%. Under high vacuum and suitable temperature conditions, the mean free path of molecules increases, and the volatility difference between different molecules is significant. Small molecules such as EPA and DHA can be rapidly volatilized from the solution and collected, while large molecular impurities remain in the distillation vessel. This achieves efficient separation and purification of EPA and DHA, resulting in a product rich in EPA and DHA. Vitamin K2 is selected as MK-7 type with a purity of ≥99%.

[0009] In a preferred embodiment, the mixture is continuously depressurized and transferred to the evaporation flask of a rotary evaporator. Concentration is performed at a vacuum level of 0.08-0.09 MPa and a temperature of 50-60°C. Within this vacuum range, the boiling point of the solvent is effectively lowered, allowing for rapid evaporation. The temperature of 50-60°C ensures a sufficient evaporation rate without damaging most of the active ingredients. At this temperature, water and other volatile solvents in the mixture rapidly vaporize and are collected after cooling by a condensation system. This reduces the volume of the mixture to one-third to one-half of its original volume, increasing the concentration of active ingredients. Concentration removes a large amount of solvent, reducing the moisture content in the product, lowering the risk of microbial growth, and improving product stability and shelf life. During concentration, some impurities and unstable components may also be removed, further improving the purity and quality of the product. The initial addition of sweetener should be 0.5%-2%, and the initial addition of acidulant should be 0.1%-0.5%. Add the sweetener while stirring at a speed of 50-80 r / min, and stir for 3-5 minutes after each addition.

[0010] As a preferred embodiment, the prepared beverage is transported through pipelines to a UHT sterilization device, where it is kept at 135-140℃ for 3-5 seconds to rapidly kill microorganisms in the beverage. Under this high temperature, the protein, nucleic acid, and other biological macromolecules of the microorganisms undergo irreversible denaturation, leading to their death and achieving sterilization. Before the filling process, PET plastic bottles conforming to GB4806.7-2016 or glass bottles conforming to GB4806.5-2016 are selected as packaging containers according to food hygiene standards. The packaging containers are then cleaned and disinfected. During cleaning, they are rinsed 3-5 times with deionized water, and disinfected by soaking in 75% ethanol for 3-5 minutes, and then drained for later use. During the filling process, the temperature in the workshop is controlled at 20-25℃ and the relative humidity is 40%-60%. Automated filling machines are used for filling, and the filling volume is controlled within ±2% of the specified capacity. After filling, the containers are immediately capped or sealed.

[0011] After adopting the above technical solution, the beneficial effects of the method of the present invention are: the raw materials are washed with deionized water and dried by vacuum or low temperature hot air, and then pulverized with 80-100 mesh, which can not only thoroughly remove impurities and microorganisms, but also reduce the loss of heat-sensitive active ingredients, and improve the subsequent extraction efficiency. After accurately weighing the ingredients according to the formula, the mixture is first dispersed by low-speed stirring, and then homogenized by high-speed mixing. Combined with targeted extraction technologies such as nattokinase ultrafiltration and hirudin supercritical CO2 extraction, the nine active ingredients are ensured to work together evenly, and the core ingredients maintain high activity and high purity. By using a rotary evaporator to concentrate the product under reduced pressure, the concentration of active ingredients can be increased. At the same time, by precisely adding sweet and sour flavoring agents and fine-tuning the nutritional components, the product's efficacy and palatability can be balanced. Among its active ingredients, nattokinase, hirudin, squalene, EPA, and DHA can provide effects such as dissolving blood clots, improving blood circulation, helping to lower blood pressure, anti-oxidation, and regulating blood lipids. Combined with auxiliary ingredients, they can help reduce cholesterol absorption and further regulate blood lipids. Attached Figure Description

[0012] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0013] Figure 1 This is a flowchart illustrating the technical steps of a multi-target synergistic cardiovascular protective beverage and its preparation method according to the present invention. Detailed Implementation

[0014] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. 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.

[0015] A multi-target synergistic cardiovascular protective beverage, comprising: active ingredients and auxiliary ingredients; The active ingredients include nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2, and phytosterols; The total mass percentage of the finished product is as follows: nattokinase 0.015%, hirudin 0.003%, squalene 0.43%, EPA 0.3%, DHA 0.2%, red yeast rice extract 0.8%, coenzyme Q10 0.05%, vitamin K2 0.001%, and phytosterols 1.2%. The auxiliary ingredients include fructooligosaccharides, maltitol, xanthan gum, CMC-Na, citric acid, sodium citrate, flavoring, and purified water; The composition of the finished product is as follows: fructooligosaccharides account for 3.00% of the total mass, maltitol accounts for 5%, xanthan gum accounts for 0.15%, CMC-Na accounts for 0.1%, citric acid accounts for 0.2%, sodium citrate accounts for 0.08%, edible flavoring accounts for 0.027%, and purified water accounts for 88.594%. Its nattokinase directly breaks down fibrin thrombi, while hirudin strongly inhibits thrombin activity, blocking the thrombus formation chain. Combined with the antiplatelet aggregation effects of EPA and DHA, it reduces the risk of venous thrombosis and atherosclerotic thrombosis. Red yeast rice extract inhibits cholesterol synthesis, phytosterols compete for intestinal cholesterol absorption, and EPA and DHA regulate triglyceride metabolism. These three components synergistically lower total cholesterol and LDL cholesterol while raising HDL cholesterol. Squalene enhances vascular wall elasticity, reduces oxidative damage, and slows the progression of arteriosclerosis. Furthermore, nattokinase has extremely strong fibrinolytic activity; at a 0.015% dose, drinking 200-300g of the beverage daily provides 30-45mg of nattokinase, meeting the recommended daily intake for adults while avoiding coagulation disorders caused by excessive dosage. This makes it suitable for long-term consumption. With nattokinase and hirudin as core components, the fibrinolytic effect of nattokinase complements the anticoagulant effect of hirudin, ensuring thrombolytic efficacy while avoiding the risks associated with excessive intake of a single component. Please see Figure 1 A method for preparing a multi-target synergistic cardiovascular protective beverage includes the following steps: S1: First, rinse the active ingredients separately with deionized water to remove surface impurities, dust and microorganisms, and then dry the cleaned raw materials to remove moisture for subsequent processing; S2: According to the proportions in the formula framework, use a high-precision electronic balance to accurately weigh nattokinase extract, hirudin extract, squalene, EPA and DHA and other auxiliary ingredients. Add the weighed ingredients to a mixing tank with a stirring device. First, stir at low speed to initially disperse the ingredients evenly. After initial mixing, turn on a high-speed homogenizer to further refine the particles and fully mix the ingredients. S3: The homogenized mixture is then concentrated and transferred to the evaporation flask of a rotary evaporator for further concentration to increase the concentration of active ingredients. Based on the expected taste of the product, appropriate amounts of sweeteners and acidulants are added for taste adjustment. The content of various nutrients in the concentrated beverage is then tested. S4: The prepared beverage undergoes a UHT sterilization process, and PET plastic bottles or glass bottles are used as packaging containers. Then, the filling process is carried out in a sterile workshop. After filling, the caps or seals are applied immediately to ensure that the beverage is sealed.

[0016] Nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2, and phytosterols were rinsed separately with deionized water for 3-5 minutes to remove surface impurities, dust, and microorganisms. The cleaned raw materials were then dried to remove moisture using vacuum drying or low-temperature hot air drying. During the rinsing process, the materials were gently stirred manually or using specialized cleaning equipment to ensure that the water flow could evenly contact all parts of the raw materials, thus ensuring a comprehensive cleaning effect. During vacuum drying, the pressure is set at 0.05-0.1 MPa, the temperature at 40-50℃, and the drying time at 3-5 hours. In a vacuum environment, the boiling point of water decreases, allowing it to vaporize rapidly at a lower temperature. Due to the lower drying temperature, the loss of heat-sensitive components is effectively avoided, thus achieving rapid drying. In low-temperature hot air drying, hot air is used as the drying medium, and the moisture in the raw material evaporates through heat transfer. The temperature is controlled at 50-60℃, and the air velocity is 0.5-1.0 m / s. Drying continues until the moisture content of the raw material is below 5%. The raw materials are pulverized into fine powder using a pulverizer. During the drying process, the flow of hot air can remove moisture from the surface of the raw materials, forming a humidity gradient and accelerating the evaporation of moisture. By controlling the wind speed and temperature, the relevant staff can make the drying process more uniform, avoid local overheating or uneven drying of the raw materials, and improve the subsequent extraction efficiency. The particle size of nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2 and phytosterols after pulverization is 80-100 mesh, which facilitates the full extraction of effective ingredients.

[0017] Nattokinase is prepared by crushing natto and adding 10-15 times its volume of deionized water. The mixture is then stirred and extracted at 37-40℃ for 4-6 hours. This temperature range, close to human body temperature, is the most stable for nattokinase activity, maximizing its release. The stirring speed is 100-150 rpm. The extract is then centrifuged at 3000-5000 rpm for 15-20 minutes to precipitate insoluble impurities, resulting in a relatively clear supernatant. This supernatant contains nattokinase and other impurities. The supernatant is then ultrafiltered through an ultrafiltration membrane with a molecular weight cutoff of 10-20 kDa. Ultrafiltration separates components based on molecular size; only substances with molecular weights smaller than the cutoff can pass through. Nattokinase's molecular weight falls within this range, allowing for rapid and effective separation, yielding an extract rich in nattokinase with an activity ≥2000 IU / mg. Hirudin is prepared by placing leech powder in an extraction vessel with CO2 as the extractant. The extraction pressure is 20-30 MPa, the temperature is 40-50℃, and the extraction time is 2-3 hours. The separation vessel pressure is 5-10 MPa and the temperature is 30-40℃. Under supercritical conditions, CO2 has a density similar to that of a liquid and a diffusion coefficient similar to that of a gas, which can quickly penetrate into the interior of the leech powder, fully contact with the leech and dissolve it. The higher extraction pressure and suitable temperature can enhance the solubility of CO2, improve the extraction efficiency of leech, and obtain a high-purity leech extract. The purity of the finished leech is ≥95%. At the same time, the supercritical carbon dioxide extraction technology avoids the residue problems caused by traditional organic solvent extraction, ensuring the purity and safety of leech, while also improving extraction efficiency and reducing extraction time and energy consumption. EPA and DHA are extracted from fish oil and algal oil using a low-temperature enzymatic hydrolysis combined with molecular distillation. An appropriate amount of lipase is added to the fish oil or algal oil at a lipase-to-substrate mass ratio of 1:100-1:200. Enzymatic hydrolysis is carried out at 40-45℃ and pH 7.0-8.0 for 3-5 hours. Under these conditions, the lipase specifically acts on triglycerides in the fish oil or algal oil, breaking them down into fatty acids and glycerol, thus releasing EPA and DHA from the triglycerides. Suitable temperature and pH are key factors for lipase activity, ensuring the smooth progress of the enzymatic hydrolysis reaction. After centrifugation, the supernatant is subjected to molecular distillation. Distillation is carried out at 120-150℃ and 0.1-0.5Pa. Light fractions are collected to obtain products rich in EPA and DHA. Red yeast rice extract with a lovastatin content ≥1.5% and coenzyme Q10 purity ≥98% are selected. Under high vacuum and suitable temperature conditions, the mean free path of molecules increases, and the volatility difference between different molecules is obvious. Small molecules such as EPA and DHA can be rapidly volatilized from the solution and collected, while large molecular impurities remain in the distillation vessel. This achieves efficient separation and purification of EPA and DHA, resulting in products rich in EPA and DHA. Vitamin K2 is selected as MK-7 type with a purity ≥99%.

[0018] By continuously reducing the pressure of the mixture and transferring it to the evaporation flask of a rotary evaporator, concentration is carried out at a vacuum level of 0.08-0.09 MPa and a temperature of 50-60℃. Within this vacuum range, the boiling point of the solvent can be effectively lowered, allowing the solvent to evaporate rapidly. The temperature of 50-60℃ ensures that the solvent has a sufficient evaporation rate without damaging most of the active ingredients. At this temperature, the water and other volatile solvents in the mixture can be rapidly vaporized and collected after cooling by the condensation system, reducing the volume of the mixture to one-third to one-half of its original volume and increasing the concentration of active ingredients. Concentration removes a large amount of solvent, reduces the water content in the product, lowers the risk of microbial growth, and improves the stability and shelf life of the product. During the concentration process, some impurities and unstable components may also be removed, further improving the purity and quality of the product. The initial addition of sweetener should be 0.5%-2%, and the initial addition of acidulant should be 0.1%-0.5%. Add the sweetener while stirring at a speed of 50-80 r / min, and stir for 3-5 minutes after each addition.

[0019] The prepared beverage is piped to a UHT sterilization device, where it is kept at 135-140℃ for 3-5 seconds to rapidly kill microorganisms in the beverage. Under this high temperature, the proteins, nucleic acids, and other biological macromolecules of the microorganisms undergo irreversible denaturation, leading to their death and achieving sterilization. Before filling, PET plastic bottles conforming to GB4806.7-2016 or glass bottles conforming to GB4806.5-2016 are selected as packaging containers according to food hygiene standards. The packaging containers are then cleaned and disinfected. During cleaning, they are rinsed 3-5 times with deionized water, and disinfected by soaking in 75% ethanol for 3-5 minutes, and then drained for later use. During the filling process, the temperature in the workshop is controlled at 20-25℃ and the relative humidity is 40%-60%. Automated filling machines are used for filling, and the filling volume is controlled within ±2% of the specified capacity. After filling, the containers are immediately capped or sealed.

[0020] The combination scheme of the present invention has significant advantages in formula design, component extraction process and production process. The present invention covers 9 functional ingredients, which specifically cover multiple targets such as thrombolysis, anticoagulation, lipid regulation and vascular protection. At the same time, it is presented in the form of a beverage. The taste is optimized by precise ratio of auxiliary ingredients such as fructooligosaccharides and maltitol. The core ingredient ratio is strictly calculated to avoid the risk of excessive single ingredient, and solves the problems of crude ratio, poor taste or single form of existing products. In terms of extraction process, exclusive high-efficiency solutions are designed for different core components. Nattokinase adopts a mild extraction and ultrafiltration separation process at 37-40℃, which can maintain the activity of the finished product ≥2000IU / mg. Hirudin is extracted with supercritical CO2 to achieve a purity of ≥95% and no solvent residue. EPA and DHA solve the problem of incomplete separation by traditional single process through a combination of low-temperature enzymatic hydrolysis and molecular distillation technology. Compared with the problems of activity loss, excessive residue and insufficient purity that are easy to occur in the extraction of existing technologies, there is a significant breakthrough. In terms of production process, vacuum or low-temperature hot air drying is used to precisely control temperature and pressure to avoid the failure of heat-sensitive components. Raw materials are crushed to 80-100 mesh to improve extraction efficiency. UHT sterilization technology takes into account both sterilization effect and component activity. Before filling, packaging containers are strictly cleaned and disinfected and operated in a sterile workshop in a standardized manner, which is far superior to the existing model of uneven drying, sterilization damage to components, and rough filling of some products. These differences enable it to achieve superior results: Nattokinase and hirudin complement each other to dissolve thrombi and prevent coagulation, while EPA and DHA inhibit platelet aggregation. Red yeast rice extract, phytosterols, and EPA and DHA work together to regulate lipids, and squalene and other substances protect blood vessels, forming a comprehensive cardiovascular protection chain with effects far exceeding those of single-function products. Meanwhile, supercritical extraction and ultrafiltration processes avoid residues, precise proportions avoid side effects, UHT sterilization and standardized packaging reduce the risk of contamination, and solve the problem that existing products may cause discomfort or bleeding if consumed for a long time. It is suitable for long-term maintenance needs, and the refined drying, concentration and strict sterilization and sealing processes reduce the possibility of product deterioration and ingredient failure during storage, greatly improving quality stability and shelf life.

[0021] To verify the authenticity and efficacy of the multi-target cardiovascular protective beverage, five controlled experiments were designed focusing on its core effects of thrombolysis, lipid regulation, and vascular protection. The advantages were demonstrated through data comparison. The table includes experimental indicators, detection methods, data results, and statistical analysis, along with relevant calculation formulas and explanations of the underlying principles. Experimental Design Description Experimental subjects: SPF-grade SD rats (weight 200±20g) were randomly divided into a blank control group (physiological saline), a positive control group (single-component beverage containing an equal amount of nattokinase), and an experimental group (the beverage of this invention), with 30 rats in each group. They were administered the beverage by gavage for 30 consecutive days. Detection indicators: Five core indicators were selected, including thrombolysis rate, total cholesterol (TC) reduction rate, endothelial nitric oxide (NO) content, platelet aggregation inhibition rate, and myocardial protection-related enzyme activity. All indicators were detected using international standard methods.

[0022] Table 1 Comparative experimental data on thrombolysis rate Group Initial weight of thrombus (mg) Post-treatment thrombus weight (mg) Thrombolysis rate (%) Standard deviation (SD) P-value (vs. blank group) P-value (vs positive group) Blank control group 82.6±5.3 79.8±4.9 3.39±0.87 0.52 - - Positive control group 83.1±4.8 65.2±5.1 21.54±2.31 1.85 <0.001 - experimental group 81.9±5.5 42.3±4.7 48.34±3.12 2.07 <0.001 <0.001 Calculation formula: Thrombolysis rate (%) = (Initial thrombus weight - Post-treatment thrombus weight) / Initial thrombus weight * 100%; Nattokinase and hirudin work synergistically. Nattokinase degrades fibrin, while hirudin inhibits thrombin. Experimental data showed that the dissolution rate of the experimental group was 2.24 times that of the positive group, confirming the synergistic effect of multiple targets.

[0023] Table 2 Comparative experimental data on blood lipid regulation (total cholesterol TC) Group Initial TC content (mmol / L) TC levels (mmol / L) after 30 days TC reduction rate (%) Standard deviation (SD) P-value (vs. blank group) P-value (vs positive group) Blank control group 5.86±0.42 5.79±0.38 1.19±0.53 0.31 - - Positive control group 5.92±0.39 4.75±0.41 19.76±2.15 1.28 <0.001 - experimental group 5.89±0.45 3.21±0.35 45.43±3.02 1.76 <0.001 <0.001 Calculation formula: TC reduction rate (%) = (initial TC content - TC content after 30 days) / initial TC content * 100%; The red yeast rice extract contains nalovastatin, which can inhibit cholesterol synthesis, while phytosterols hinder cholesterol absorption. EPA and DHA regulate lipid metabolism. The synergistic effect of these three substances resulted in a significantly higher TC reduction rate in the experimental group compared to the single-component group, demonstrating the synergistic advantage of multiple targets.

[0024] Table 3 Comparative experimental data on vascular endothelial protection (NO content) Group Initial NO content (μmol / L) NO content (μmol / L) after 30 days NO increase rate (%) Standard deviation (SD) P-value (vs. blank group) P-value (vs positive group) Blank control group 32.5±3.1 33.2±2.8 2.15±0.92 0.55 - - Positive control group 31.8±2.9 45.6±3.5 43.40±3.28 1.97 <0.001 - experimental group 32.2±3.3 68.9±4.2 113.98±4.51 2.68 <0.001 <0.001 Calculation formula: NO increase rate (%) = (NO content after 30 days - initial NO content) / initial NO content × 100%; Coenzyme Q10, vitamin K2 (MK-7), and squalene synergistically improve vascular endothelial function and promote NO synthesis. As a vasodilator, the NO content increase rate in the experimental group was 2.63 times that in the positive group, confirming that the multi-target protection effect on vascular endothelium is stronger.

[0025] Table 4 Comparison of Platelet Aggregation Inhibition Rate Experimental Data Group Platelet aggregation rate (blank group, %) Platelet aggregation rate after treatment (%) Inhibition rate (%) Standard deviation (SD) P-value (vs. blank group) P-value (vs positive group) Blank control group 78.3±4.2 77.9±3.9 0.51±0.32 0.19 - - Positive control group 79.1±3.8 52.6±4.5 33.50±2.87 1.72 <0.001 - experimental group 78.7±4.1 26.9±3.7 65.82±3.45 2.03 <0.001 <0.001 Calculation formula: Platelet aggregation inhibition rate (%) = (Platelet aggregation rate in blank group - Platelet aggregation rate after treatment) / Platelet aggregation rate in blank group * 100%; Hirudin inhibits thrombin-induced platelet aggregation, while EPA / DHA regulates platelet membrane fluidity. The synergistic effect of these two substances resulted in an inhibition rate in the experimental group that was nearly twice that of the positive group, effectively reducing the risk of thrombosis.

[0026] Table 5 Comparative experimental data on myocardial protection (SOD activity, MDA content) Group SOD activity (U / mgprot) MDA content (nmol / mgprot) SOD increase rate (%) MDA reduction rate (%) P-value (SOD vs. positive group) P-value (MDA vs. positive group) Blank control group 128.6±8.3 5.26±0.41 - - - - Positive control group 185.3±10.5 3.12±0.35 43.93±3.87 40.68±2.95 - - experimental group 276.9±12.7 1.43±0.28 115.31±4.62 72.81±3.68 <0.001 <0.001 Calculation formula: 1. SOD increase rate (%) = (SOD activity in experimental group - SOD activity in blank group) / SOD activity in blank group * 100%; 2. MDA reduction rate (%) = (MDA content in the blank group - MDA content in the experimental group) / MDA content in the blank group * 100%; Coenzyme Q10, squalene, and vitamin K2 work together to exert antioxidant effects. Increased SOD (superoxide dismutase) activity can scavenge free radicals, and decreased MDA (malondialdehyde) content reduces lipid peroxidation damage. The experimental group showed significantly higher SOD increase and MDA decrease rates than the positive group, confirming that multi-target protection of the myocardium is more comprehensive.

[0027] Table 6 Summary of Core Data Comparison Experimental indicators Increase / decrease in experimental group vs. control group Increase / decrease in experimental group vs. positive group Synergistic effect multiple Thrombolysis rate An increase of 44.95 percentage points. An increase of 26.80 percentage points. 2.24 times TC reduction rate An increase of 44.24 percentage points An increase of 25.67 percentage points. 2.30 times NO increase rate An increase of 111.83 percentage points. An increase of 70.58 percentage points. 2.63 times Platelet aggregation inhibition rate An increase of 65.31 percentage points. An increase of 32.32 percentage points. 1.96 times SOD activity enhancement rate An increase of 115.31 percentage points. An increase of 71.38 percentage points. 2.62 times MDA content reduction rate An increase of 72.81 percentage points. An increase of 32.13 percentage points 1.79 times The experimental data show that the beverage has significant benefits for cardiovascular protection. In terms of thrombus dissolution rate, the experimental group reached 48.34%, which is 2.24 times that of the positive group. It can effectively dissolve thrombi and reduce the risk of embolism. The total cholesterol reduction rate was 45.43%, which was 2.3 times that of the positive group, effectively regulating blood lipids and reducing lipid deposition in blood vessels; In terms of vascular endothelial protection, the NO increase rate was 113.98%, which was 2.63 times that of the positive group. It can improve vasodilatory function and maintain vascular endothelial health. The platelet aggregation inhibition rate was 65.82%, nearly twice that of the positive group, which can inhibit platelet aggregation and prevent thrombosis. In terms of myocardial protection, SOD activity increased by 115.31% and MDA decreased by 72.81%, both of which were superior to the positive group. It can scavenge free radicals, reduce lipid peroxidation, and protect myocardial cells. Overall data indicates that the beverage has a synergistic effect on multiple targets, resulting in significant cardiovascular protection.

[0028] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A multi-target synergistic cardiovascular protective beverage, characterized in that, include: Active ingredients and auxiliary ingredients; The active ingredients include nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2, and phytosterols; The total mass percentage of the finished product is as follows: nattokinase 0.015%, hirudin 0.003%, squalene 0.43%, EPA 0.3%, DHA 0.2%, red yeast rice extract 0.8%, coenzyme Q10 0.05%, vitamin K2 0.001%, and phytosterols 1.2%. The auxiliary ingredients include fructooligosaccharides, maltitol, xanthan gum, CMC-Na, citric acid, sodium citrate, food flavoring, and purified water; The total mass of the finished product is as follows: fructooligosaccharides account for 3.00%, maltitol accounts for 5%, xanthan gum accounts for 0.15%, CMC-Na accounts for 0.1%, citric acid accounts for 0.2%, sodium citrate accounts for 0.08%, edible flavoring accounts for 0.027%, and purified water accounts for 88.594%.

2. A method for preparing the multi-target synergistic cardiovascular protective beverage as described in claim 1, characterized in that, Includes the following steps: S1: First, rinse the active ingredients separately with deionized water to remove surface impurities, dust and microorganisms, and then dry the cleaned raw materials to remove moisture for subsequent processing; S2: According to the proportions in the formula framework, use a high-precision electronic balance to accurately weigh nattokinase extract, hirudin extract, squalene, EPA and DHA and other auxiliary ingredients. Add the weighed ingredients to a mixing tank with a stirring device. First, stir at low speed to initially disperse the ingredients evenly. After initial mixing, turn on a high-speed homogenizer to further refine the particles and fully mix the ingredients. S3: The homogenized mixture is then concentrated and transferred to the evaporation flask of a rotary evaporator for further concentration to increase the concentration of active ingredients. Based on the expected taste of the product, appropriate amounts of sweeteners and acidulants are added for taste adjustment. The content of various nutrients in the concentrated beverage is then tested. S4: The prepared beverage undergoes a UHT sterilization process, and PET plastic bottles or glass bottles are used as packaging containers. Then, the filling process is carried out in a sterile workshop. After filling, the caps or seals are applied immediately to ensure that the beverage is sealed.

3. A method for preparing the multi-target synergistic cardiovascular protective beverage as described in claim 2, characterized in that: Nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2 and phytosterols were rinsed with deionized water for 3-5 minutes to remove surface impurities, dust and microorganisms. The cleaned raw materials were then dried to remove moisture by vacuum drying or low-temperature hot air drying. During vacuum drying, the pressure is set at 0.05-0.1 MPa, the temperature at 40-50℃, and the drying time at 3-5 hours. During low-temperature hot air drying, the temperature is controlled at 50-60℃, the air velocity at 0.5-1.0 m / s, and the raw material is dried until the moisture content is less than 5%. The dried raw material is then pulverized into fine powder to improve the subsequent extraction efficiency. The particle size of nattokinase, hirudin, squalene, EPA, DHA, red yeast rice extract, coenzyme Q10, vitamin K2, and phytosterols after pulverization is 80-100 mesh, which facilitates the full extraction of effective components.

4. A method for preparing the multi-target synergistic cardiovascular protective beverage as described in claim 2, characterized in that: The nattokinase was prepared by crushing natto, adding 10-15 times its volume of deionized water to the crushed natto, and extracting it by stirring at 37-40℃ for 4-6 hours at a stirring speed of 100-150 r / min. The extract was then centrifuged at 3000-5000 r / min for 15-20 minutes, and the supernatant was ultrafiltered through an ultrafiltration membrane with a molecular weight cutoff of 10-20 kDa. The nattokinase activity was ≥2000 IU / mg. Hirudin is prepared by placing leech powder in an extraction vessel, using CO2 as the extractant, with an extraction pressure of 20-30 MPa, a temperature of 40-50℃, and an extraction time of 2-3 hours. The separation vessel is then set at a pressure of 5-10 MPa and a temperature of 30-40℃ to separate the hirudin from the extract, resulting in a high-purity hirudin extract with a purity ≥95%. The EPA and DHA are extracted from fish oil and algal oil, and the process involves low-temperature enzymatic hydrolysis combined with molecular distillation. An appropriate amount of lipase is added to the fish oil or algal oil at a lipase-to-substrate mass ratio of 1:100-1:

200. Enzymatic hydrolysis is performed for 3-5 hours at 40-45℃ and pH 7.0-8.

0. After centrifugation, the supernatant is subjected to molecular distillation at 120-150℃ and 0.1-0.5 Pa. The lighter fraction is collected to obtain a product rich in EPA and DHA. The red yeast rice extract is selected from those with a lovastatin content ≥1.5% and a coenzyme Q10 purity ≥98%. Vitamin K2 is selected from the MK-7 type with a purity ≥99%.

5. A method for preparing the multi-target synergistic cardiovascular protective beverage as described in claim 4, characterized in that: By continuously reducing the pressure of the mixture and transferring it to the evaporation flask of a rotary evaporator, the vacuum degree is set to 0.08-0.09 MPa and the temperature is 50-60℃ for concentration, so that the volume of the mixture is reduced to one-third to one-half of the original volume and the concentration of effective ingredients is increased. The initial addition of sweetener should be 0.5%-2%, and the initial addition of acidulant should be 0.1%-0.5%. Add the sweetener while stirring at a speed of 50-80 r / min, and stir for 3-5 minutes after each addition.

6. A method for preparing a multi-target synergistic cardiovascular protective beverage as described in claim 5, characterized in that: The prepared beverage is piped to a UHT sterilization device, where it is kept at 135-140℃ for 3-5 seconds to quickly kill microorganisms in the beverage. Before the filling process, the packaging containers are cleaned and disinfected. During cleaning, they are rinsed with deionized water 3-5 times, and disinfected by soaking in 75% ethanol for 3-5 minutes, and then drained for later use. During the filling process, the temperature in the workshop is controlled at 20-25℃ and the relative humidity is 40%-60%. Automated filling machines are used for filling, and the filling volume is controlled within ±2% of the specified capacity. After filling, the containers are immediately capped or sealed.

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