Preparation method and device of cocoa product capable of improving cardiovascular function

Abstract

The invention discloses a preparation method and device of a cocoa product with a cardiovascular function improving function, and belongs to the technical field of food processing. Dry cocoa beans and blueberry berries are used as raw materials, the dry cocoa beans are crushed and sieved and then mixed with deionized water to extract cocoa butter, then the cocoa butter is mixed with wall-broken blueberry homogenate, the deionized water is used as a guiding agent to aggregate reaction components, cells are completely broken through a three-layer stepped reaction layer, and the cocoa butter is obtained. The dissolution of cocoa butter in the cocoa beans and active substances in the blueberries is promoted; the whole reaction device and the conveying pipeline are negative-pressure and oxygen-free, so that the stability of active substances is ensured. The cocoa product obtained by adopting the device and the method disclosed by the invention is fine and smooth in taste, melts in the mouth, is basically free from greasy feeling and granular feeling, perfectly retains the special flavors of cocoa beans and blueberries, is rich in cocoa butter and anthocyanin substances, and has relatively strong effects of preventing atherosclerosis and improving cardiovascular.

Description

Technical Field

[0001] The present invention relates to the technical field of food processing. In particular, it relates to a preparation method and device for a cocoa product with improved cardiovascular function. Background Art

[0002] At present, cardiovascular health problems have become increasingly prominent. According to statistics, the incidence and mortality of cardiovascular diseases continue to rise. Functional foods can play a certain role in improving cardiovascular health, such as reducing cholesterol, cleaning the "garbage" on the blood vessel wall, reducing vascular inflammatory responses, keeping blood vessels unobstructed, and promoting the normal operation of the cardiovascular system. It is a major solution for improving cardiovascular health problems.

[0003] Among them, the flavanol substances in cocoa beans have powerful antioxidant effects, which can effectively scavenge free radicals in the body, reduce oxidative stress, inhibit the expression and release of various inflammatory factors, and at the same time help reduce the inflammatory burden on the arterial wall; the substances extracted from cocoa beans can improve vascular function and promote the production of nitric oxide. Nitric oxide is an important vasodilator that can relax vascular smooth muscle and lower blood pressure. In addition, it can also reduce vascular tension by inhibiting the activity of angiotensin-converting enzyme (ACE). This effect helps to improve the diastolic function of blood vessels, increase blood flow, and reduce the risk of vascular stenosis and obstruction. However, in the processing of traditional cocoa products, due to high-temperature treatment or improper extraction, these beneficial components are often lost in large quantities, thus reducing their cardiovascular-improving effects.

[0004] Moreover, blueberries are rich in anthocyanin substances and also have powerful antioxidant properties. In the human body, free radicals are constantly generated. When there are too many free radicals, they will attack vascular endothelial cells, trigger a series of inflammatory responses, promote lipid deposition on the blood vessel wall, and thus accelerate the process of cardiovascular diseases. The anthocyanins in blueberries can effectively scavenge free radicals, reduce the damage caused by oxidative stress to blood vessels, inhibit inflammatory responses, and prevent lipid peroxidation, thereby reducing the risk of cardiovascular diseases. The nutrients in blueberries can improve vascular endothelial function, make blood vessel dilation smoother, maintain normal blood pressure levels, reduce the pressure on the blood vessel wall when blood flows in the blood vessels, and further play a preventive effect on atherosclerosis by regulating blood lipid metabolism and reducing the content of low-density lipoprotein cholesterol in the blood and reducing cholesterol accumulation on the blood vessel wall.

[0005] Therefore, combining blueberries with cocoa beans can further enhance the antioxidant capacity and cardiovascular health benefits of functional foods. However, during the processing of traditional cocoa products and blueberry products, problems such as loss of active ingredients, poor taste, and residue of harmful substances often occur. For example, in the invention patent with the application number CN201911137689.1, cocoa is pressed to remove oil, then degreased with n-hexane, extracted with acetone-aqueous solution, filtered, and the precipitate is dried under reduced pressure at 60 °C to obtain cocoa fat-soluble polyphenols, and the filtrate is dried in one step to obtain the non-fat-soluble part. Another invention patent with the application number CN201710110527.3 extracts cocoa by heating with ethanol water, purifies it through macroporous resin, and then dries it to obtain the product. The chromatography column method used for refining the extracts of these inventions and the organic solvents used are likely to leave residues, which can easily pose safety hazards. Moreover, flavanols will decompose at high temperatures, resulting in low content of active ingredients, poor water solubility, reduced nutritional value, and being difficult to be accepted by consumers. Another example is that in the invention patent with the application number CN201210559011.4, an anthocyanin-containing solution is obtained by alcohol dissolution and heating extraction. However, anthocyanins are sensitive to heat, and improper operation can easily lead to their loss.

[0006] Therefore, when preparing cocoa products by combining cocoa with blueberries, how to retain the active ingredients of both simultaneously and endow the product with good taste and texture is a technical problem. Summary of the Invention

[0007] Technical Problem

[0008] Currently, there is no device for co-extracting cocoa butter and anthocyanins on the market, and there are also problems with separate extraction: organic solvents are used as extraction media and are likely to leave residues and are difficult to remove; anthocyanins have poor stability and are prone to inactivation; reaction conditions are difficult to control and the extraction rate is low; the continuity level of the production process is low.

[0009] Technical Solution

[0010] To solve the above problems, the present invention overcomes the disadvantages and deficiencies of the prior art and provides a preparation method and production device for cocoa products with improved cardiovascular function, which have a high level of production continuity, high production efficiency and conform to the green concept. Using cocoa beans and blueberries as raw materials, the dry cocoa beans are crushed and sieved, then mixed with deionized water to extract cocoa butter, and then mixed with the homogenate of broken blueberries. Using deionized water as a guiding agent, the reaction components are aggregated, and the cells are completely broken through a three-layer stepped reaction layer to promote the dissolution of cocoa butter in cocoa beans and active substances in blueberries; the entire reaction device and the filtrate conveying pipeline are under negative pressure and anaerobic, ensuring the activity of soluble substances. In the method, the crushed dry cocoa beans are in full contact with the blueberry homogenate for continuous reaction. After the reaction, the mixture enters a microwave-enzymolysis treatment tower, undergoes a three-layer stepped reaction and then filtration. The filtrate enters the subsequent freeze-drying process, and the filter residue is mixed with deionized water again, ultrasonically treated and then filtered for the second time. The filtrate returns to the dry cocoa bean and blueberry mixing reactor and replaces deionized water as the reaction solvent for the subsequent dry cocoa bean powder, while the filter residue is reused for the next time.

[0011] The first object of the present invention is to provide a production device for efficiently and continuously producing cocoa products with improved cardiovascular function. The production device consists of a dry cocoa bean storage tank 1, a blueberry storage tank 2, a deionized water storage tank 3, a crusher 4, a cell breaker 5, an ultrasonic filtration device 6, a double-screw stirrer 7, a microwave-enzymolysis treatment tower 8, a filtration device 9, a filtrate storage tank 10, a molding machine 11, a pre-cooling machine 12, a freeze-dryer 13, and a vacuum pump 14.

[0012] The dry cocoa bean storage tank 1 communicates with the crusher 4; the blueberry storage tank 2 communicates with the cell breaker 5;

[0013] Automatic switches are installed at the bottoms of the crushing device 4 and the cell breaker 5, and a sieve and a filter screen are respectively placed under the baffles.

[0014] The deionized water storage tank 3 communicates with the ultrasonic filtration device 6 and the double-screw stirrer 7; an automatic switch is installed in the middle of the ultrasonic filtration device 6, and a filter screen is placed under the baffle; an ultrasonic device is installed in the upper part of the baffle.

[0015] The crusher 4, the cell breaker 5, the deionized water storage tank 3 and the ultrasonic filtration device 6 communicate with the double-screw stirrer 7;

[0016] The double-screw stirrer 7 communicates with the microwave-enzymolysis treatment tower 8;

[0017] In the middle of the microwave-enzymolysis treatment device 8 is an immobilized enzyme layer 18; above the immobilized enzyme layer 18 is a pulsed microwave field, on which a magnetron array 15 and a rotating and tiltable tray 16 are arranged;

[0018] The magnetron array 15 is arranged on the top of the microwave-enzymatic hydrolysis treatment device 8; the rotatable and tiltable tray 16 is located below the magnetron array 15 and is fixed by a support rod perpendicular to the immobilized enzyme membrane layer 8.

[0019] There are automatically openable baffles above and below the immobilized enzyme layer 8; below the immobilized enzyme layer 8 is an ultrasonic cavitation field, provided with a spiral deflector 19, and the deflector descends in a spiral shape around the inner wall of the device, and a dual-frequency transducer is arranged on the inner wall.

[0020] The microwave-enzymatic hydrolysis treatment device 8 communicates with the filtration device 9, and the filtration device communicates with the ultrasonic filtration device 6 and the filtrate storage tank 10.

[0021] The filtrate storage tank 10 communicates with the mold filling machine 11, and then communicates with the precooler 12 and the freeze dryer 13 in sequence.

[0022] Vacuum pumps 14 are provided between the cell wall breaking device 5, the ultrasonic device 6, the microwave-enzymatic hydrolysis treatment device 8 and the double-screw agitator 7, between the microwave-enzymatic hydrolysis treatment device 8, the ultrasonic filtration device 6 and the filtration device 9, and between the filtration device 9 and the filtrate storage tank 10.

[0023] Furthermore, the magnetron array is composed of annular magnetrons.

[0024] Furthermore, the power of the magnetron is 2 - 2.5 kW and the frequency is 2000 - 2500 MHz.

[0025] Furthermore, the tray of the rotatable and tiltable tray is a concave tray, and the depth of its groove is 6 - 10 mm.

[0026] Furthermore, the immobilized enzyme layer is composed of an adsorption material; the adsorption material includes one or more of sponge and chitosan.

[0027] Furthermore, pectinase and cellulase are loaded in the immobilized enzyme layer, and the total enzyme loading is 30 - 35 mg / g.

[0028] Furthermore, the mass ratio of pectinase to cellulase is 3 - 5:1.

[0029] Furthermore, the enzyme activity of pectinase is 18000 - 22000 U / g, and the enzyme activity of cellulase is 2000 - 4000 U / g.

[0030] Furthermore, the power density of the dual-frequency transducer is 50 - 70 W / L and the frequency is 20 - 40 kHz.

[0031] Furthermore, the mesh number of the filter screen in the filtration device is 200 - 400 meshes.

[0032] Furthermore, the interconnected parts in the production device are connected by conveying pipelines; a negative pressure and oxygen-free environment is maintained in the conveying pipelines.

[0033] The second object of the present invention is to provide a preparation method of a cocoa product with improved cardiovascular function, and the method is prepared by using the above-mentioned production device for efficiently and continuously producing a cocoa product with improved cardiovascular function. The specific steps of the method are as follows:

[0034] Crush and sieve dry cocoa beans in a crusher to obtain dry cocoa bean powder. Input the dry cocoa bean powder into a double-helix stirrer, add water, and extract cocoa butter in the double-helix stirrer under a vacuum and anaerobic environment. Then, homogenize and mix the obtained cocoa butter with blueberry jam that has been broken by a blender in the double-helix stirrer to obtain a material. Then, input the material into a microwave-enzymatic treatment tower, and pass through a pulsed microwave field, an immobilized enzyme layer, and an ultrasonic cavitation field from top to bottom. Filter the treated material through a filtering device to obtain a primary filtrate and a primary filter residue. Then, pour the primary filtrate into a mold by a molding machine, pre-cool and solidify it in a pre-cooling machine, and finally freeze-dry it in a freeze-dryer to obtain a cocoa product with improved cardiovascular function.

[0035] Further, the sieve mesh number of the dry cocoa bean powder is 100 - 300 meshes.

[0036] Further, the mass ratio of the dry cocoa bean powder to water is 1:18 - 20.

[0037] Further, the vacuum degree of the vacuum and anaerobic environment is 0.03 - 0.05 MPa.

[0038] Further, the reaction temperature for extracting cocoa butter is 5 - 10 °C, and the reaction time is 1 - 3 h.

[0039] Further, the mass ratio of the blueberry jam to the dry cocoa bean powder is 1:2.5 - 3.

[0040] Further, the temperature for homogenizing and mixing the blueberry jam and cocoa butter is 5 - 8 °C, and the time is 4 - 5 h.

[0041] Preferably, the temperature for homogenizing and mixing the blueberry jam and cocoa butter is 5 - 6 °C.

[0042] Further, the frequency in the pulsed microwave field is 2000 - 2500 MHz.

[0043] Further, the treatment time in the pulsed microwave field is 5 - 10 min.

[0044] Further, the treatment time of the immobilized enzyme layer is 20 - 30 min, and the temperature is 6 - 10 °C.

[0045] Further, the power in the ultrasonic cavitation field is 0.3 - 0.7 w / cm 2 .

[0046] Further, the treatment time in the ultrasonic cavitation field is 5 to 10 minutes.

[0047] Further, the size of the mold used for casting is 0.4 to 0.6 × 0.4 to 0.6 cm.

[0048] Further, the pre-freezing temperature is -30 to -10 °C.

[0049] Further, the primary filter residue is added with water in a mass ratio of 1:4 to 6, and treated in an ultrasonic filtration device at an ultrasonic condition of 4 to 6 °C and 300 to 500 W for 10 to 30 minutes, and then filtered again to obtain a secondary filtrate and a secondary filter residue; the secondary filtrate can be used to replace water as the reaction solvent for dry cocoa bean powder, and the secondary filter residue can be combined with the primary filter residue and reused.

[0050] The present invention provides a cocoa product with improved cardiovascular function prepared according to the above method.

[0051] The application of the cocoa product provided by the present invention in the fields of food and health products.

[0052] Further, the health product is a health product that helps to maintain a healthy level of blood lipids (cholesterol / triglyceride).

[0053] Beneficial effects

[0054] In the present invention, by adopting the processes of stepwise crushing, continuous reaction, microwave-enzymolysis treatment and filter residue recycling, the biological activity of soluble substances in cocoa beans is maximally retained in a negative pressure and anaerobic environment, the dissolution efficiency of cocoa butter and the enrichment amount of anthocyanins are significantly improved, and at the same time, the freeze-drying technology endows the cocoa product with a delicate taste and low greasiness characteristics; the continuous production mode therein combines microwave-enzymolysis treatment, ultrasonic-assisted extraction and filter residue recycling technology, which not only improves the utilization rate of raw materials, reduces energy consumption and waste discharge, but also strengthens the health effect of the product in improving cardiovascular through the synergistic effect of functional components, realizing the efficient and green manufacturing of functional foods and the simultaneous optimization of sensory quality. Brief description of the drawings

[0055] Figure 1 is a schematic diagram of the production device for preparing a cocoa product with improved cardiovascular function in Example 1 of the present invention;

[0056] In the figure: 1 dry cocoa bean storage tank, 2 blueberry storage tank, 3 deionized water storage tank, 4 crusher, 5 cell wall breaker, 6 ultrasonic filtration device, 7 double-screw stirrer, 8 microwave-enzymolysis treatment tower, 9 filtration device, 10 filtrate storage tank, 11 casting machine, 12 pre-cooling machine, 13 freeze-dryer, 14 vacuum pump.

[0057] In the figure: ① to ⑧ are reaction steps; ① After the dry cocoa beans are crushed and sieved, they are mixed with deionized water in a double - helix stirrer; ② When the dry cocoa beans and blueberries are mixed for 3 h in step ①, the homogenized blueberry pulp after cell wall breaking is introduced into the double - helix stirrer; ③ After the reaction in the double - helix stirrer ends, the mixture enters the microwave - enzymatic hydrolysis treatment tower and passes through the upper - layer pulsed microwave field, the middle - layer immobilized enzyme membrane, and the lower - layer ultrasonic cavitation field from top to bottom in sequence; ④ After the reaction in step ③ ends, the mixture is filtered; ⑤ The filtrate obtained from the filtration in step ④ enters the filtrate storage tank, and then is subjected to molding, pre - cooling, and freeze - drying; ⑥ The filter residue obtained from the filtration in step ④ is introduced into the ultrasonic device, and deionized water is introduced simultaneously; ⑦ After the ultrasonic treatment in step ⑥ ends, it is filtered; ⑧ The filtrate obtained from the filtration in step ⑦ is introduced into the double - helix stirrer to replace the addition of deionized water in step ①.

[0058] Figure 2 It is a sectional view of the microwave - enzymatic hydrolysis treatment device 8 in Example 1;

[0059] In the figure: 15 is a magnetron array, 16 is a rotatable and tiltable tray, 17 is an automatically opened baffle, 18 is an immobilized enzyme layer, and 19 is a spiral guide plate.

[0060] Figure 3 It shows the content changes of cocoa butter, flavanols, and anthocyanins in cocoa products under different substrate mass ratios (cocoa beans: blueberries) in Example 3 of the present invention.

[0061] Figure 4 It is the content changes of cocoa butter, flavanols, and anthocyanins in cocoa products under different reaction times in Example 5 of the present invention.

[0062] Figure 5 It is the content changes of cocoa butter, flavanols, and anthocyanins in cocoa products under different mesh numbers of the filter screen in Example 6 of the present invention.

[0063] Figure 6 It is the content changes of cocoa butter, flavanols, and anthocyanins in cocoa products under different vacuum degrees in Example 7 of the present invention.

[0064] Figure 7 It is the content changes of cocoa butter, flavanols, and anthocyanins in cocoa products under different reaction temperatures in Example 8 of the present invention.

[0065] Figure 8 It is the content changes of cocoa butter, flavanols, and anthocyanins in the cocoa products of Example 2 and Comparative Examples 1, 2, 3, and 4 of the present invention.

[0066] Figure 9 It is the effect diagram of the influence of the cocoa product in Example 2 of the present invention on cholesterol. Detailed implementation manners

[0067] For the convenience of understanding the present invention, the present invention will be described more comprehensively below in conjunction with embodiments and drawings. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. On the contrary, these embodiments are provided to make the understanding of the disclosure of the present invention more thorough and comprehensive.

[0068] The materials, reagents, instruments and methods used in the present invention, unless otherwise specified, are conventional materials, reagents, instruments and methods in the art and can be obtained through commercial channels and existing methods.

[0069] (1) Analysis and detection of cocoa butter in cocoa products

[0070] Weigh 4 mg of cocoa products; add 4 mL of n-hexane, then add 2 mL of 0.5 mol / L KOH methanol solution, and reflux in a 70 °C water bath for 30 min. Take it out and cool, transfer it to a graduated test tube, add deionized water to 10 mL, shake, ultrasonicate, centrifuge, take the supernatant, add a small amount of anhydrous sodium sulfate, and take the dried sample for GC-MS analysis.

[0071] Gas chromatography analysis conditions: Chromatographic column DB-FFAP (60 m × 0.25 mm × 0.25 μm) elastic quartz capillary column. Temperature programming: Initial temperature 100 °C is maintained for 5 min, heated to 240 °C at a rate of 10 °C / min, and maintained for 10 min; injection volume is 1 μL; carrier gas (He) flow rate is 1 mL / min; splitless; solvent delay is 6 min.

[0072] Mass spectrometry conditions: Electron impact (EI) ion source; injection port temperature is 240 °C; ion source temperature is 230 °C; quadrupole temperature is 150 °C; multiplier voltage is 1376 V; electron energy is 70 eV; emission current is 34.6 μA; interface temperature is 240 °C; mass scan range is 20 - 500 amu.

[0073] Calculation of cocoa butter content: Accurately weigh different masses of cocoa butter standards, dissolve them with n-hexane and prepare a series of standard solutions with different concentrations. According to the above chromatographic conditions, inject the standard solutions for analysis in sequence, and record the peak areas of the characteristic peaks of cocoa butter in each standard solution. Taking the concentration of the standard solution as the abscissa and the peak area as the ordinate, plot a standard curve to obtain a linear regression equation. Inject the processed sample solution and record the peak area of the characteristic peak of cocoa butter in the sample. According to the linear regression equation of the standard curve, calculate the concentration of cocoa butter in the sample solution, and then combine the dilution factor and the mass of the sample weighed to calculate the content of cocoa butter in the cocoa products.

[0074] (2) Analysis and detection of flavanols in cocoa products

[0075] Weigh 5 g of cocoa products into a 50 mL centrifuge tube, add 45 mL of n-hexane, and vortex for 1 min. Place the centrifuge tube in a 50 °C water bath and sonicate for 5 min. After taking it out, centrifuge at 3000 rpm for 5 min. Discard the n-hexane layer. Repeat the above operation twice to remove oil impurities. Accurately weigh 1 g of defatted sample into a 15 mL centrifuge tube, add 5 mL of extraction solution (acetonitrile: water: acetic acid = 70:29.5:0.5), vortex at medium speed for 2 min to extract flavanols, and after centrifuging at 3000 rmp for 5 min, the supernatant is analyzed by high performance liquid chromatography.

[0076] High performance liquid chromatography analysis conditions: Develosil Diol 250×4.6 mm (5 μm particle size) chromatographic column. Column temperature 35 °C, flow rate 1 mL / min, injection volume 5 μL, and the auto-injection temperature is set and maintained at 5 °C. Mobile phase: binary gradient, composed of acidic acetonitrile [(A) CH3CN–HOAc, 98 + 2 (v / v)] and acidic methanol aqueous solution [(B) CH3OH–H2O–HOAc, 95 + 3 + 2 (v / v / v)]. The initial mobile phase is 7% B and is isocratic for 3 min. Subsequently, the proportion of solvent B is increased to 37.6% within 57 min, and then increased to 100% B within 3 min. Maintain at 100% B for 7 min, and then return to the initial conditions (7% B) to equilibrate for 6 min.

[0077] Fluorescence detection (FLD) conditions: excitation wavelength 230 nm, emission wavelength 321 nm.

[0078] Calculation of flavanol content: Prepare a series of standard solutions with different concentrations using flavanol standards, draw a standard curve, and obtain a linear regression equation. where C is the concentration of flavanols in the sample extract calculated according to the standard curve (mg / mL), V is the total volume of the extract (mL), and m is the mass of the cocoa product sample weighed (g). Determine in parallel three times and calculate the average value.

[0079] (3) Determination of anthocyanin content in cocoa products

[0080] Weigh 5 g of the cocoa product sample into a 50 mL centrifuge tube, add 45 mL of n-hexane, vortex at medium speed for 1 min to mix them thoroughly. Place the centrifuge tube in a 50 °C constant temperature water bath and sonicate for 20 min (sonication power 500 W) to further promote the dissolution of the oil. Then centrifuge at 3000 rpm for 5 min to separate the n-hexane layer from the cocoa product residue, and discard the upper n-hexane layer. Repeat this operation twice to ensure that the oil is fully removed. Accurately weigh 1 g of the sample from the defatted cocoa product residue and transfer it to a 15 mL centrifuge tube. Add 5 mL of the extraction solution (acidified methanol: water = 80:20, v / v, adjusted to pH 2.5 with hydrochloric acid), vortex at medium speed for 2 min to allow the sample to come into full contact with the extraction solution and extract the anthocyanins, and then analyze by liquid chromatography-mass spectrometry.

[0081] High performance liquid chromatography analysis conditions: Chromatographic column: Agilent ZORBAX SB C-18 (250×4.6 mm, 5 μm); Mobile phase: Phase A is 0.5% formic acid aqueous solution, Phase B is pure acetonitrile. Elution program: 0 - 5 min: 100% A, 5 - 20 min: 100% - 90% A, 20 - 40 min: 90% - 87% A, 40 - 44 min: 87% - 80% A, 44 - 50 min: 80% - 75% A, 50 - 55 min: 75% - 100% A; Detection wavelength: 280 nm; Flow rate: 1 mL / min; Column temperature: 25 °C; Injection volume: 20 μL.

[0082] Mass spectrometry conditions: Electrospray ionization source; Simultaneous positive and negative ion scanning; Mass scanning range m / z: 100 - 1000; Dry gas flow rate: 5 L / min; Dry gas temperature: 325 °C; Nebulizer pressure: 45 psi; Sheath gas temperature: 350 °C; Sheath gas flow rate: 11 mL / min; Capillary voltage: 3000 V in positive ion mode and 3500 V in negative ion mode.

[0083] Calculation of anthocyanin content: Prepare a standard curve to obtain the linear regression equation. Where C is the concentration of anthocyanins in the sample extract calculated according to the standard curve (mg / mL), V is the total volume of the extract (mL), and m is the mass of the cocoa product sample weighed (g); Determine in parallel three times and calculate the average value.

[0084] Example 1: A production device for a cocoa product with improved cardiovascular function

[0085] As Figure 1As shown in the figure, the production device of a cocoa product with improved cardiovascular function consists of a dry cocoa bean storage tank 1, a blueberry storage tank 2, a deionized water storage tank 3, a crusher 4, a cell wall breaker 5, an ultrasonic filtration device 6, a double - helix stirrer 7, a microwave - enzymolysis treatment tower 8, a filtration device 9, a filtrate storage tank 10, a molding machine 11, a pre - cooler 12, a freeze - dryer 13, and a vacuum pump 14.

[0086] The dry cocoa bean storage tank 1 is connected to the crusher 4, and the dry cocoa beans in the storage tank can be input into the crusher to be crushed into dry cocoa bean powder; the blueberry storage tank 2 is connected to the cell wall breaker 5, and the blueberries in the tank can be input into the cell wall breaker to be broken into blueberry jam.

[0087] The bottom of both the crushing device 4 and the cell wall breaker 5 are equipped with automatically - opening baffles, and a sieve and a filter screen are respectively placed under the baffles. When the crushing or cell wall breaking of dry cocoa beans or blueberries stops, the baffles will open, and the dry cocoa bean powder or blueberry jam can be automatically sieved or filtered.

[0088] The deionized water storage tank 3 is connected to the ultrasonic filtration device 6 and the double - helix stirrer 7; the ultrasonic filtration device 6 is provided with an automatically - opening baffle in the middle, and a filter screen is placed under the baffle; the baffle divides the ultrasonic filtration device into upper and lower parts. The upper part is the ultrasonic part for ultrasonic dispersion of materials, and the lower part is the liquid storage part. When the ultrasonic treatment ends, the baffle will automatically open, and the ultrasonic - treated liquid can pass through the filter screen for filtration, and then the obtained filtrate enters the liquid storage part for standing.

[0089] The crusher 4, the cell wall breaker 5, the deionized water storage tank 3, and the ultrasonic filtration device 6 are connected to the double - helix stirrer 7, and the cocoa powder, blueberry jam, deionized water, and filtrate can be input into the stirrer to be stirred to obtain a mixture; the double - helix stirrer 7 is connected to the microwave - enzymolysis treatment tower 8, and the obtained mixture can be input into the treatment tower.

[0090] The microwave - enzymolysis treatment device 8 is as Figure 2 shown, and it is provided with a magnetron array 15, a rotatable and tiltable tray 16, an automatically - opening baffle 17, an immobilized enzyme layer 18, and a spiral deflector 19; the immobilized enzyme layer is in the middle of the treatment device, dividing the treatment device into upper, middle, and lower layers. The upper layer is a pulsed microwave field, the middle layer is the immobilized enzyme layer, and the lower layer is an ultrasonic cavitation field;

[0091] A magnetron array 15 and a rotatable and tiltable tray 16 are arranged in the pulsed microwave field; the magnetron array 15 is composed of annular magnetrons, each magnetron having a power of 2.5 kW and a frequency of 2450 MHz; the annular magnetrons are arranged on the top of the processing device, and the number can be adjusted according to the size of the processing device; the rotatable and tiltable tray 16 is located below the magnetron array 15 and is fixed by a support rod perpendicular to the immobilized enzyme membrane layer 18. A rotating shaft is provided between the support rod and the tray to enable the tray to rotate and tilt; the tray is a concave tray with a groove depth of 8 mm, and the surface of the tray is coated with a food-grade silicon carbide coating; the heights of the rotatable and tiltable trays decrease in sequence, and the size and number of the trays can be adjusted by itself.

[0092] The immobilized enzyme layer 18 is composed of an adsorption material (chitosan), and the adsorption material is loaded with pectinase and cellulase, and the mass ratio of the two is 4:1, and the total loading amount is 32 mg / g; there are automatically openable baffles above and below the immobilized enzyme layer. After the treatment of the material in the pulsed microwave field, the upper baffle opens and the lower baffle closes. When the liquid material fills the immobilized enzyme layer, the upper baffle closes, and the liquid material undergoes enzymatic hydrolysis treatment. The enzymatic hydrolysis time is 20 min. After the enzymatic hydrolysis ends, the lower baffle opens. Among them, both pectinase and cellulase are purchased from Zhongchen Biotechnology, and the enzyme activities are 20000 U / g and 3000 U / g respectively.

[0093] A spiral deflector 19 is arranged in the ultrasonic cavitation field. The deflector spirally surrounds the inner wall of the device and descends to extend the time for the material to pass through the ultrasonic cavitation field; a dual-frequency transducer is arranged on the inner wall of the ultrasonic cavitation field, with a power density of 60 W / L and a working frequency of 28 kHz + 40 kHz; a laser turbidity sensor can be installed at the water outlet of the microwave-enzymatic hydrolysis treatment device.

[0094] The microwave-enzymatic hydrolysis treatment device 8 communicates with the filtration device 9, and the treated mixture can be passed into the filtration device for filtration; the filtration device communicates with the ultrasonic filtration device 6 and the filtrate storage tank 10. The filter residue obtained by filtration is input into the ultrasonic filtration device, and the obtained filtrate is input into the filtrate storage tank.

[0095] The filtrate storage tank 10 communicates with the mold filling machine 11, and then communicates with the precooling machine 12 and the freeze dryer 13 in sequence. The filtrate can be processed by the mold filling machine, the precooling machine and the freeze dryer to obtain the final product.

[0096] Vacuum pumps 14 are provided between the cell wall breaking device 5, the ultrasonic device 6, the microwave-enzymatic hydrolysis treatment device 8 and the double helix stirrer 7, between the microwave-enzymatic hydrolysis treatment device 8, the ultrasonic device 6 and the filtration device 9, and between the filtration device 9 and the filtrate storage tank 10.

[0097] Example 2: A preparation method of a cocoa product with improved cardiovascular function

[0098] It is carried out using the production device of Example 1. Using roasted dry cocoa beans and fresh blueberries as raw materials, the dry cocoa beans are crushed and sieved (200 mesh) in a crusher to obtain dry cocoa bean powder. The dry cocoa bean powder is input into a double - helix stirrer, and deionized water (which can be replaced by secondary filtrate) is added at 18 times the mass of the dry cocoa bean powder. Cocoa butter is extracted in the double - helix stirrer under a vacuum and anaerobic environment, where the vacuum degree is 0.05 MPa, the reaction temperature is 6 °C, and the reaction time is 2 h. Then, the obtained cocoa butter and blueberry jam that has been broken by a blender are homogenized and mixed in the double - helix stirrer. The mass of the blueberry jam is 3 times that of the dry cocoa bean powder, and the continuous reaction is carried out at 5 °C for 5 h. After completion, the obtained material is input into a microwave - enzymatic hydrolysis treatment tower, and it passes through a pulsed microwave field (2450 MHz, 10 min), an immobilized enzyme layer (the ratio of pectinase / cellulase is 4:1, the enzymatic hydrolysis time is 20 min, and the enzymatic hydrolysis temperature is 8 °C), and an ultrasonic cavitation field (28 KHz + 40 KHz, 0.5 w / cm 2 , 10 min) from top to bottom. Subsequently, the treated material is filtered through a filtering device (300 mesh) to obtain a primary filtrate and a primary filter residue. The primary filtrate is molded in a molding machine through a 0.5×0.5 cm mold, pre - cooled and solidified at - 20 °C in a pre - cooler, and freeze - dried in a freeze - dryer to obtain cocoa products.

[0099] The primary filter residue is added with deionized water at a mass ratio of 1:5, and treated in an ultrasonic filtering device under ultrasonic conditions of 5 °C and 400 W for 20 minutes, and then filtered twice (300 mesh) to obtain a secondary filtrate and a secondary filter residue. The secondary filtrate can be used to replace deionized water as the reaction solvent for dry cocoa bean powder, and the secondary filter residue can be combined with the primary filter residue for reuse.

[0100] After the obtained cocoa products are tested for content, the cocoa butter content is 31.27%, the flavanol content is 1.79%, and the anthocyanin content is 1.18%.

[0101] Example 3: Determination of the substrate mass ratio

[0102] To determine the optimal substrate mass ratio, it is carried out according to the steps of Example 2, where only the mass ratio of dry cocoa bean powder and blueberry jam is changed. Specifically, in terms of mass ratio, the ratios of dry cocoa bean powder and blueberry jam are 1:1, 1:2, 1:4, 1:5, and 1:6 respectively, and the cocoa butter, flavanol, and anthocyanin contents in the obtained cocoa products are detected.

[0103] The obtained results are as Figure 3As shown, when the mass ratio of dry cocoa bean powder to blueberry jam changes from 1:1 to 1:3, the contents of cocoa butter, flavanols, and anthocyanins all increase significantly. However, when the mass ratio of dry cocoa bean powder to blueberry jam continues to increase, the contents of cocoa butter and flavanols decrease significantly. This result indicates that a mass ratio of 1:3 of dry cocoa bean powder to blueberry jam is the ideal substrate mass ratio for preparing cocoa products with improved cardiovascular function.

[0104] Example 4: Determination of deionized water content

[0105] To determine the optimal addition amount of deionized water (secondary filtrate) in cocoa butter, the steps of Example 2 were followed, where only the addition amount of deionized water added when extracting cocoa butter using a double - helix stirrer was adjusted. Specifically, based on the mass of dry cocoa bean powder, 12 times, 14 times, 16 times, and 20 times the amount of deionized water was added, and the touch and taste of the obtained cocoa products were evaluated according to Table 1.

[0106] The obtained results are shown in Table 2, which indicates that 18 times is the ideal water addition amount for preparing cocoa products with improved cardiovascular function.

[0107] Table 1 Sensory attribute evaluation criteria

[0108]

[0109]

[0110] Table 2 Evaluation results of cocoa products with different water contents in Example 4

[0111]

[0112] Example 5: Determination of reaction time

[0113] To determine the optimal reaction time conditions, the steps of Example 2 were followed, where only the homogenization mixing time of dry cocoa bean powder and blueberry jam in the double - helix stirrer was adjusted. Specifically, with the temperature kept constant, the reaction times were 3 h, 7 h, 9 h, and 11 h respectively, and the contents of cocoa butter, flavanols, and anthocyanins in the obtained cocoa products were detected.

[0114] The obtained results are as Figure 4 shown. As the reaction time increases from 3 h to 5 h, the contents of cocoa butter, flavanols, and anthocyanins increase significantly. After 3 h, the contents of flavanols and anthocyanins do not increase, and at the same time, the increase in the content of cocoa butter is not obvious. This result indicates that 5 h is the ideal reaction time for preparing cocoa products with improved cardiovascular function.

[0115] Example 6: Determination of sieve mesh number

[0116] To determine the optimal mesh number for filtering the mixed reactants, the steps of Example 2 were followed, with only the mesh number of the filter screen in the filtering device being adjusted, specifically: 80 mesh, 100 mesh, 200 mesh, and 400 mesh, and the contents and taste of cocoa butter, flavanols, and anthocyanins in the obtained cocoa products were detected.

[0117] The results obtained are as Figure 5 shown in and Table 3. It can be found that the taste and nutritional value of the cocoa products obtained using a 300-mesh sieve are the best.

[0118] Table 3 Evaluation Results of Cocoa Products with Different Sieve Meshes in Example 5

[0119]

[0120]

[0121] Example 7: Determination of Vacuum Degree

[0122] To determine the optimal vacuum degree for the reaction, the steps of Example 2 were followed, with only the vacuum degree during the extraction of cocoa butter using a double-screw agitator being adjusted, specifically: 0.06 MPa, 0.07 MPa, 0.08 MPa, and 0.09 MPa, and the contents of cocoa butter, flavanols, and anthocyanins in the obtained cocoa products were detected.

[0123] The results obtained are as Figure 6 shown. As the vacuum degree increases, the contents of cocoa butter, flavanols, and anthocyanins all gradually decrease. This result indicates that 0.05 MPa is the ideal reaction vacuum degree for preparing cocoa products with improved cardiovascular function.

[0124] Example 8: Determination of Reaction Temperature

[0125] To determine the optimal reaction temperature conditions, the steps of Example 2 were followed, with only the temperature of the dry cocoa bean powder and blueberry jam during homogenization and mixing in the double-screw agitator being adjusted, specifically: 2 °C, 8 °C, 11 °C, and 15 °C, and the contents of cocoa butter, flavanols, and anthocyanins in the obtained cocoa products were detected.

[0126] The results obtained are as Figure 7 shown. As the temperature rises, the cocoa butter content gradually increases, but when the reaction temperature is above 5 °C, the flavanol content decreases, and when the reaction temperature is above 7 °C and continues to increase, the anthocyanin content decreases. Considering comprehensively, 6 °C is the optimal reaction temperature for preparing cocoa products with improved cardiovascular function.

[0127] Example 9

[0128] (1) The cocoa products prepared in Example 2 were formulated into a cocoa product culture solution according to the following steps. The specific process is as follows:

[0129] S1, grinding the cocoa product prepared in Example 2 through a 300-mesh sieve to obtain cocoa product powder;

[0130] S2, dissolving 1% (w / v) soy lecithin in preheated (37°C) pH 3.5 citric acid buffer (protected from light), stirring magnetically until clear, adding cocoa product powder, vortexing for preliminary mixing, and cocoa product mother liquor (cocoa product concentration is 5wt%);

[0131] S3, then the cocoa product mother liquor was ultrasonically treated in an ice bath (300 W, pulse mode, 5 minutes), and then high-pressure homogenized (500 bar, cycled 3 times) to make the particle size of the cocoa product powder therein ≤200 nm;

[0132] S4. After filtering the cocoa product mother liquor through a 0.22 μm filter membrane, add it into DMEM culture medium containing 10% FBS to obtain cocoa product culture fluids with concentrations of 0.2, 0.4, 0.6, 0.8, and 1 mg / mL, respectively, and store in the dark for later use.

[0133] (2) Take flavanol powder and blueberry anthocyanin powder and prepare a culture solution according to the following steps, wherein the flavanol powder is purchased from Yolanda Biofactory Enterprise Store, and the blueberry anthocyanin powder is purchased from Missha Biotechnology. The specific process is as follows:

[0134] a. The flavanol powder and blueberry anthocyanin powder were dissolved in preheated (37°C) pH 3.5 citric acid buffer and vortexed until completely dissolved to obtain a flavanol mother solution (flavanol concentration of 5 wt%) and a blueberry anthocyanin mother solution (blueberry anthocyanin concentration of 5 wt%);

[0135] b. Take the flavanol mother solution and blueberry anthocyanin mother solution and add them to DMEM medium containing 10% FBS, mix gently, and obtain concentration gradients of 0.2, 0.4, 0.6, 0.8, 1 mg / mL flavanol culture medium and anthocyanin culture medium;

[0136] c. Filter and sterilize the flavanol culture solution and anthocyanin culture solution (0.22 μm filter membrane) to obtain the flavanol culture solution and anthocyanin culture solution, and store them away from light for later use.

[0137] (3) The three cell culture fluids obtained above were used for cell experiments. The specific process was as follows:

[0138] 1. Recovery:

[0139] Take out the cryopreserved THP-1 cells from the liquid nitrogen tank, place them in a 37°C water bath for about 3 minutes, transfer the melted cell suspension to 10 mL of complete medium under a laminar flow hood, mix well by dilution, and centrifuge at 600 - 800 rpm for 3 minutes. Remove the supernatant medium, resuspend the cells with 5 mL of complete medium, transfer them to a T25 culture flask, and culture them upright in an incubator. The culture conditions are 37°C and 5% CO2. The THP-1 cells are purchased from the ACTT cell bank.

[0140] 2. Subculture:

[0141] Half-medium change subculture: Resuspend the medium in the cell culture flask filled with THP-1 cells and transfer it to a new T25 culture flask at a ratio of 1:3, and supplement the complete medium to 5 mL and culture it in an incubator; Full-medium change subculture: Collect the cell medium, centrifuge at 600 - 800 rpm for 3 minutes, remove the supernatant, resuspend the cells with the complete medium, and transfer them to a T25 culture flask at a ratio of 1:3 and culture it in an incubator.

[0142] 3. Macrophage differentiation:

[0143] Seed the THP-1 cells at a density of 1×10 5 cells / well in a culture plate, add 100 ng / mL PMA, and culture at 37°C and 5% CO2 for 48 hours. Replace the fresh medium without PMA and let the cells adhere and mature for 24 hours.

[0144] 4. Cell treatment and foam formation induction:

[0145] Divide the differentiated macrophages into a cocoa product group, a flavanol group, and an anthocyanin group. The corresponding compound addition concentrations in each group are divided into five concentrations: 0 mg / mL, 0.2 mg / mL, 0.4 mg / mL, 0.6 mg / mL, and 0.8 mg / mL. The added compounds are the cocoa product cell culture medium, flavanol, and anthocyanin cell culture medium prepared according to the above steps. Pretreat the grouped cells, that is, add the medium containing different concentrations of compounds, incubate for 24 h, then add ox-LDL (50 μg / mL), and continue to culture for 48 hours.

[0146] 5. Cell viability analysis:

[0147] Seed the logarithmic-phase THP-1 cells at 8×10 4 / well in a 96-well plate. After inducing with PMA for 48 h, remove the original medium, replace it with drugs at different intervention concentrations, and treat the cells for 24 h. Prepare the medium containing CCK8 reagent at a ratio of 1:10, continue to culture for 2 h after changing the medium, and set up a cell-free blank group. Use an enzyme-linked immunosorbent assay (ELISA) reader to measure the absorbance at 450 nm. Calculate the cell viability according to the instructions of the kit, with five parallels in each group.

[0148] 6. Analysis of lipid content and cholesterol content:

[0149] Oil Red O staining: THP-1 cells in the logarithmic growth phase were seeded in 24-well plates. The culture medium was discarded, and the cells were washed three times with PBS. Then, 500 μL of 4% paraformaldehyde was added to fix the cells for 20 min, after which the fixative was removed. The cells were washed three times with PBS, and then 500 μL of Oil Red O dye was added to stain the cells for 30 min, after which the dye was removed. The cells were washed three times with PBS and then observed under a microscope for lipid droplets. 500 μL of isopropanol was added to each well to dissolve the lipids, and an appropriate amount was taken and placed in a 96-well plate. The absorbance at 540 nm was measured using a microplate reader to semi-quantify the lipid content, with three parallels in each group.

[0150] 7. Cholesterol detection:

[0151] THP-1 cells in the logarithmic growth phase were seeded in 6-well plates at a density of 2×10 6 / well. After inducing culture with PMA for 48 h, a model was established under different drug interventions. According to the methods described in the cell TC detection kit and cell FC detection kit instructions, the total cholesterol content and free cholesterol content in the cells were detected. The cholesterol ester content = total cholesterol content - free cholesterol content. The protein concentration of each sample was quantified using a BCA kit, with three parallels in each group.

[0152] The results are shown in Figure 9 and Table 4.

[0153] Table 4

[0154]

[0155] Applying Jin Zhengjun's Q-value calculation formula, the effect (E A ) of the flavanol culture solution was 23%, the effect (E B ) of the anthocyanin culture solution was 31%, and the effect (E A+B ) of the cocoa product culture solution was 57%. It was calculated that Q > 1.15. Therefore, there is a synergistic relationship between flavanols and anthocyanins in the cocoa products of the present invention, that is, the cocoa products prepared from dry cocoa beans and blueberry berries can better and more effectively improve cardiovascular diseases.

[0156]

[0157] E A and E B are the individual effects of A and B, and E A+B is the combined effect of A and B. Q < 0.85 indicates an antagonistic effect between A and B; 0.85 ≤ Q ≤ 1.15 indicates an additive effect between A and B; Q ≥ 1.15 indicates a synergistic effect between A and B.

[0158] Comparative Example 1

[0159] Same as Example 2, except that in the microwave-enzymolysis treatment device, the pulsed microwave field and the ultrasonic cavitation field are not turned on, and enzymolysis is directly carried out on the immobilized enzyme layer for 0 h to obtain Figure 8 the content change results of cocoa butter, flavanol and anthocyanin. The results show that traditional single-stage extraction makes the leaching of cocoa butter, flavanol and anthocyanin incomplete, and the extraction rate decreases. Vacuum cold extraction can further increase the content of these three substances in cocoa products.

[0160] Comparative Example 2

[0161] Same as Example 2, except that when using a double-screw stirrer to extract cocoa butter, the solvent used is n-hexane at 18 times the dry cocoa powder, and ethanol at 15 times the blueberries is used to extract anthocyanin to obtain Figure 8 the content change results of cocoa butter, flavanol and anthocyanin. The results show that the alcohol extraction method limits the dissolution of cocoa butter and anthocyanin, and at the same time, the product has a bitter taste. Considering comprehensively, the mixed reaction is more suitable for the dissolution of cocoa butter and soluble active substances.

[0162] Comparative Example 3

[0163] Same as Example 2, except that an atmospheric pressure aerobic environment is adopted to obtain Figure 8 the content change results of cocoa butter, flavanol and anthocyanin. The results show that substances such as flavanol and anthocyanin are destroyed under the atmospheric pressure aerobic environment, and the extraction rate drops significantly. At the same time, the cocoa products have an oxidized rancid taste of oil. Adopting a negative pressure anaerobic environment can ensure the activity of water-soluble active substances and at the same time inhibit the oxidation and spoilage of oil.

[0164] The embodiments provided above are not intended to limit the scope covered by the present invention, nor are the described steps intended to limit their execution order. Obvious improvements made by those skilled in the art to the present invention in combination with the existing common general knowledge also fall within the protection scope defined by the claims of the present invention.

Claims

1. An apparatus for continuously and efficiently producing a cocoa product with improved cardiovascular function, characterized in that, The production device consists of a dry cocoa bean storage tank, a blueberry storage tank, a deionized water storage tank, a crusher, a cell wall breaker, an ultrasonic filtration device, a twin-screw agitator, a microwave-enzymolysis treatment tower, a filtration device, a filtrate storage tank, a molding machine, a precooler, a freeze dryer, and a vacuum pump; The dry cocoa bean storage tank is connected to the crusher; the blueberry storage tank is connected to the cell wall breaker; the deionized water storage tank is connected to the ultrasonic filtration device and the twin-screw agitator; the crusher, the cell wall breaker, and the ultrasonic filtration device are connected to the twin-screw agitator; the twin-screw agitator is connected to the microwave-enzymolysis treatment tower; The middle of the microwave-enzymolysis treatment device is an immobilized enzyme layer; above the immobilized enzyme layer is a pulsed microwave field, where a magnetron array and a rotatable and tiltable tray are arranged; The magnetron array is arranged at the top of the microwave-enzymolysis treatment device; the rotatable and tiltable tray is located below the magnetron array and is fixed by a support rod perpendicular to the immobilized enzyme membrane layer; Below the immobilized enzyme layer is an ultrasonic cavitation field, which is provided with a spiral deflector. The deflector spirally descends around the inner wall of the device, and a dual-frequency transducer is arranged on the inner wall; The microwave-enzymolysis treatment device is connected to the filtration device, and the filtration device is connected to the ultrasonic filtration device and the filtrate storage tank; The filtrate storage tank is connected to the molding machine, and then the molding machine, the precooler, and the freeze dryer are connected in sequence; Vacuum pumps are provided between the cell wall breaking device, the ultrasonic device, the microwave-enzymolysis treatment device and the twin-screw agitator, between the microwave-enzymolysis treatment device, the ultrasonic filtration device and the filtration device, and between the filtration device and the filtrate storage tank; The connected parts are connected by conveying pipelines; a negative pressure and oxygen-free environment is maintained in the conveying pipelines.

2. The production device according to claim 1, characterized in that, The magnetron array consists of annular magnetrons; the power of the magnetrons is 2 - 2.5 kW, and the frequency is 2000 - 2500 MHz.

3. The production device according to claim 1, characterized in that, The immobilized enzyme layer is loaded with pectinase and cellulase, and the total enzyme loading amount is 30 - 35 mg / g; the mass ratio of pectinase to cellulase is 3 - 5:1; the enzyme activity of pectinase is 18000 - 22000 U / g, and the enzyme activity of cellulase is 2000 - 4000 U / g.

4. The production device according to claim 1, characterized in that, The power density of the dual-frequency transducer is 50 - 70 W / L, and the frequency is 20 - 40 kHz.

5. A method for preparing a cocoa product with improved cardiovascular function, characterized in that, The preparation method is carried out using the production device for efficiently and continuously producing cocoa products with improved cardiovascular function described in any one of claims 1 - 4. The specific steps of the preparation method are as follows: Crush and sieve dry cocoa beans in a crusher to obtain dry cocoa bean powder, input the dry cocoa bean powder into a twin-screw agitator, add water, and extract cocoa butter in the twin-screw agitator in a vacuum and oxygen-free environment. Then, homogenize and mix the obtained cocoa butter with blueberry jam broken by a cell wall breaker in the twin-screw agitator to obtain a material. Then, input the material into a microwave-enzymolysis treatment tower, and pass through the pulsed microwave field, the immobilized enzyme layer, and the ultrasonic cavitation field from top to bottom. Filter the treated material through a filtration device to obtain a primary filtrate and a primary filter residue. Then, mold the primary filtrate with a molding machine, perform precooling and solidification in a precooler, and finally freeze-dry in a freeze dryer to obtain cocoa products with improved cardiovascular function.

6. According to the preparation method described in claim 5, characterized in that, The sieving mesh number of the dry cocoa bean powder is 100 - 300 meshes; the mass ratio of the dry cocoa bean powder to water is 1:18 - 20; the vacuum degree of the vacuum and oxygen-free environment is 0.03 - 0.05 MPa; the reaction temperature for extracting cocoa butter is 5 - 10 °C, and the reaction time is 1 - 3 h.

7. According to the preparation method described in claim 5, characterized in that, The mass ratio of the blueberry jam to the dry cocoa bean powder is 1:2.5 - 3; the temperature for the homogeneous mixing of the blueberry jam and the cocoa butter is 5 - 8 °C, and the time is 4 - 5 h.

8. According to the preparation method described in claim 5, it is characterized in that In the pulsed microwave field, the frequency is 2000 - 2500 MHz, and the treatment time in the pulsed microwave field is 5 - 10 min; the treatment time of the immobilized enzyme layer is 20 - 30 min, and the temperature is 6 - 10 °C; in the ultrasonic cavitation field, the power is 0.3 - 0.7 w / cm 2 ; the treatment time in the ultrasonic cavitation field is 5 - 10 min.

9. A cocoa product with improved cardiovascular function, characterized in that, The cocoa product is prepared by the preparation method according to any one of claims 5 - 8.

10. Application of the cocoa product described in claim 9 in the fields of food and health products.

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