Pinus koraiensis kernel oil direct drink beneficial to cardiovascular health and preparation method of pinus koraiensis kernel oil direct drink

Abstract

The invention provides a Korean pine seed kernel oil direct drink beneficial to cardiovascular health and a preparation method thereof.The preparation method comprises the steps that Korean pine seed kernels and tea leaves are smashed and dried together, then compound enzyme is added for enzymolysis treatment, and then high-pressure extraction is conducted to obtain high-yield antioxidant Korean pine seed kernel oil; meanwhile, embedding the probiotics by using the enzymolysis whey protein, and performing secondary embedding by using the hawthorn pectin to prepare probiotic particles; the particles and zymolytic casein are added into Korean pine kernel oil together to form a stable emulsion, so that the stability, flavor and taste of the oil are improved, and the synergistic antihypertensive and cardiovascular protection effects of the ACE inhibitory peptide and the hawthorn are exerted; finally, fruit juice such as apples and pomegranates is added, so that the oxidation resistance is further enhanced, the flavor is improved, the shelf life is prolonged, and the comprehensive gain effect of the product on cardiovascular health is improved.

Description

Technical Field

[0001] This invention belongs to the field of food technology, specifically relating to a direct beverage made from red pine kernel oil that is beneficial to cardiovascular health and its preparation method. Background Technology

[0002] Korean pine kernels are rich in various nutrients, including unsaturated fatty acids (such as linoleic acid and linolenic acid), vitamins, and minerals, which have positive effects on cardiovascular health. The unsaturated fatty acids in Korean pine kernel oil can help lower LDL cholesterol levels, prevent the occurrence and development of atherosclerosis, and thus maintain cardiovascular health. In particular, the unique pinolenic acid has significant physiological activities such as lowering blood lipids and improving cardiovascular health.

[0003] Tea not only contains polyphenols, which have antioxidant properties, but also contributes to cardiovascular health, such as... Figure 1 As shown, the active substances contained in different teas have multiple functions.

[0004] Whey protein bioactive peptides possess ACE inhibitory activity, including inhibiting angiotensin-converting enzyme (ACE) activity, lowering blood pressure, inhibiting platelet aggregation, lowering cholesterol, and exhibiting antioxidant effects, thus protecting cardiovascular health. For example, bioactive peptides in whey protein, such as ACE inhibitory peptides, anti-inflammatory peptides, and antioxidant peptides, may affect the cardiovascular system through multiple mechanisms. ACE inhibitory peptides in whey protein can be combined with various food and medicinal materials to protect cardiovascular health. According to existing research, the synergistic effect of food and medicinal materials with ACE inhibitory peptides has been extensively studied, especially showing significant effects in lowering blood pressure and protecting the cardiovascular system. Firstly, food and medicinal materials such as hawthorn, cassia seed, and wolfberry have certain blood pressure-lowering effects, but their active ingredients are easily decomposed in gastric acid, and their bitterness limits their application. However, by combining them with ACE inhibitory peptides, their ACE inhibitory activity can be significantly improved, and the stability and absorption rate of the active ingredients can be enhanced. Casein is a natural protein found in milk and is widely used in the food industry due to its excellent nutritional and functional properties. Casein possesses excellent emulsifying and foam-forming abilities. Furthermore, its surface is rich in amino acid residues, which can be chemically modified to further enhance its properties, such as strengthening its interactions with sugars or amino acids. Casein adsorbs at the oil-water interface, forming an interfacial film; monosaccharides and amino acids can enhance the stability of this film through intermolecular interactions. In addition, specific peptides in casein hydrolysates (such as VPP and IPP) have shown cardiovascular health protective effects in animal models and clinical trials.

[0005] Korean pine kernels are high in unsaturated fatty acids, making them highly susceptible to oxidation and rancidity, resulting in the so-called hazelnut flavor. Furthermore, current pine nut oil extraction methods suffer from low extraction rates, and high temperatures and other physical conditions further promote oxidation. Traditional Korean pine kernel oil products suffer from poor oxidative stability, low bioavailability, and unpleasant taste, limiting their application as direct-consumption products. Korean pine kernel products that are beneficial for cardiovascular health are primarily available as additives to Korean pine kernel powder. Summary of the Invention

[0006] The technical problem to be solved: To enrich the types of Korean pine nut products, improve the oxidative stability, bioavailability, and taste of Korean pine kernel oil, and achieve certain health benefits for the cardiovascular system. This invention prepares a direct-drinking product with a Korean pine kernel oil content greater than 70% through the compounding of functional ingredients. This invention improves the oil yield and prevents oxidation through optimized oil extraction processes, and enhances the oxidative stability, bioavailability, and taste of the Korean pine kernel oil through functional emulsification of the oil. Probiotic capsules are added to improve fat metabolism after oil intake, avoiding digestive problems after consuming Korean pine kernel oil. Egg yolk lecithin and lipolytic enzymes are combined to promote bile secretion and reduce the burden of digesting fats.

[0007] Technical solution: A method for preparing a direct beverage made from red pine kernel oil that is beneficial to cardiovascular health, comprising the following steps: S1. Grinding and drying: The shelled red pine nuts are mixed with tea leaves and ground together, then placed in an oven to dry overnight, and sieved to obtain mixed red pine nut powder; S2. Extraction of Korean pine kernel oil: Add the compound enzyme aqueous solution to the mixed Korean pine nut powder and stir. Sonicate in a water bath, pack into a high-pressure bag, seal and place in a high-pressure device for high-pressure treatment. After the treatment, filter and centrifuge to collect the Korean pine kernel oil. S3. Enzymatic hydrolysis: Whey protein and casein are added to a complex enzyme buffer, pH is adjusted, and the mixture is sonicated in a water bath. The pH and sonication power are adjusted again to inactivate the enzyme. After inactivation, the pH is restored to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S4. Hawthorn pretreatment: Wash the hawthorns, remove the pits, cook them, crush them, add sugar solution again and boil them, sieve them while hot to obtain hawthorn pulp, cool them briefly and place them in a water bath; S5. Preparation of probiotic microparticles: Mix and stir the compound probiotic suspension and lecithin solution, add the enzymatically hydrolyzed whey protein solution and lipase, stir and hydrate, add to hawthorn pulp, homogenize and spray dry to obtain probiotic microparticles; S6. Ingredient Mixing: After mixing probiotic microparticles, enzymatically hydrolyzed casein solution and fruit juice, add to red pine kernel oil, homogenize at high speed and then perform microfluidic homogenization to obtain red pine kernel oil direct beverage.

[0008] Furthermore, in step S1, the mass ratio of red pine nuts to tea leaves is (50-100):1; the temperature of the drying oven is 50-60℃; and the sieve used for sieving is 10-20 mesh.

[0009] Furthermore, the tea leaves are a mixture of green tea and black tea.

[0010] Furthermore, in step S2, the compound enzyme is a mixture of pectinase, cellulase, protease, and amylase in a mass ratio of (1-2):(2-3):(1-2):1; the concentration of the compound enzyme aqueous solution is 10-20 wt.%; the mass ratio of the compound enzyme aqueous solution to the mixed red pine nut powder is (3-5):10; the temperature of the water bath ultrasound is 35-45℃, the time of the water bath ultrasound is 90-180 min, and the power of the water bath ultrasound is 200-500 W; the pressure of the high-pressure treatment is 25-45 MPa, and the time of the high-pressure treatment is 5-10 min.

[0011] Furthermore, in step S3, the complex enzyme is papain and bromelain in a mass ratio of 1:1, and the enzyme content in the complex enzyme buffer is 0.6-1 wt.%; the mass ratio of whey protein, casein and complex enzyme buffer is (1-2):10; the pH is adjusted to 6-8; the water bath temperature is 35-40℃, the ultrasonic power is 100-200W, and the ultrasonic time is 30-60min; the pH is adjusted again to 5-6, and the ultrasonic power is 500-600W; the pH is restored to 6-8.

[0012] Furthermore, in step S4, the mass ratio of hawthorn to sugar solution is (5-10):1, the concentration of sugar solution is 10-15 wt.%, the sieve used for sieving is 6 mesh, and the temperature of the water bath is 30-37℃.

[0013] Furthermore, in step S5, the compound probiotics consist of Bifidobacterium and Saccharomyces boulardii with a colony count ratio of (1-2):1, and the concentration of the compound probiotic suspension is 10¹¹ CFU / g; the concentration of the lecithin solution is 1-2 wt.%; the mass ratio of the compound probiotic suspension, lecithin solution, enzymatically hydrolyzed whey protein solution, lipase, and hawthorn pulp is 10:10:(10-20):1:(10-20); the stirring speed is 100-200 rpm, and the stirring time is 10-15 min; the stirring speed for hydration is 200-300 rpm, and the stirring time is 30-60 min, with a temperature of 37℃; the homogenization pressure is 5-10 MPa.

[0014] Furthermore, the lipase is a triglyceride hydrolase.

[0015] Furthermore, in step S6, the fruit juice is one or a combination of apple and pomegranate juice; the mass ratio of probiotic microparticles, enzymatically hydrolyzed casein solution, fruit juice and red pine kernel oil is (1-3):10:(3-6):(50-70); the high-speed homogenization speed is 8000-10000 rpm, the high-speed homogenization time is 10-15 min; and the microjet homogenization pressure is 15-20 MPa.

[0016] The above-described preparation method yields a red pine kernel oil direct beverage that is beneficial to cardiovascular health. Beneficial effects

[0017] This invention uses papain and bromelain. The hydrolysate of milk protein after enzymatic hydrolysis has high ACE inhibitory activity. The ACE inhibitory peptide is combined with hawthorn, a food and medicine homology, to inhibit ACE activity in a dual way. It can significantly enhance the antihypertensive effect through multi-target synergistic effect, while improving cardiovascular health and reducing the risk of drug side effects.

[0018] This invention uses 70% high-content red pine kernel oil as a base, creating the first direct-drinking functional beverage with red pine kernel oil as the main ingredient, filling a market gap. The probiotic double-layer encapsulated microparticles (whey protein / hawthorn pectin) ensure the colonization of live bacteria in the intestines, alleviate the burden of lipid digestion, and promote the lipolysis complex system (egg yolk lecithin + lipase) to activate bile secretion, accelerate fat metabolism, and avoid gastrointestinal discomfort.

[0019] The probiotic microparticles of this invention are protected by a phospholipid layer that provides hydrophobic protection similar to a cell membrane, a protein layer that provides physical and enzymatic barriers, and hawthorn, which is rich in pectin, that provides an acid barrier and colon-targeting ability, forming a layered protection that enhances the activity of probiotics and their tolerance to adverse environments such as acids and bile salts. Triglyceride hydrolase is added to the microparticles to assist in the hydrolysis of triglycerides, releasing the functional fatty acids of red pine kernel oil, which are then encapsulated and isolated to prevent contact with oils in the product. Furthermore, the microparticles have excellent emulsifying effects on oils, improving the dispersion stability of oils and protecting them.

[0020] In this invention, the unsaturated fatty acids in red pine kernel oil and the ACE inhibitory peptides of enzymatically hydrolyzed casein and whey protein, along with the flavonoid pectin of hawthorn and the polyphenols in the juice, work synergistically through multiple pathways to lower blood pressure and improve blood lipids, thereby enhancing the function. In addition, the juice can also harmonize the flavor and significantly improve consumer acceptance.

[0021] This invention utilizes a method of first enzymatically hydrolyzing red pine kernels and then extracting red pine kernel oil under high pressure to improve the oil yield and quality of red pine kernels. Adding tea leaves to red pine kernels before extracting oil not only plays a role in preventing oxidation and improving the flavor of the oil, but also the tea leaves have the effect of improving cardiovascular health, further enhancing the cardiovascular health function of the product. Attached Figure Description

[0022] Figure 1 A schematic diagram illustrating the role of components in green and black tea in improving cardiovascular health. Detailed Implementation

[0023] The present invention will be further described below with reference to the accompanying drawings and embodiments. The following embodiments are illustrative of the present invention, but the present invention is not limited to the following embodiments: Example 1

[0024] The extraction method for Korean pine kernel oil includes the following steps: S1. Grinding and drying: Mix 5000g of shelled red pine nuts with 50g of black tea and 50g of green tea, grind them together, put them in a 50℃ oven to dry overnight, and pass them through a 10-mesh sieve to obtain mixed red pine nut powder; S2. Extraction of Korean pine kernel oil: 20g of pectinase, 40g of cellulase, 20g of protease and 20g of amylase were dissolved in 900g of water to obtain a compound enzyme aqueous solution. This solution was then added to 2000g of mixed Korean pine nut powder and stirred. The mixture was then subjected to a 40℃ water bath and ultrasonic treatment at 300W for 120 minutes. The mixture was then placed in a high-pressure bag, sealed, and placed in a high-pressure device for high-pressure treatment at 25MPa for 5 minutes. After the treatment, the mixture was filtered and centrifuged to collect the Korean pine kernel oil. Comparative Example 1

[0025] The difference between this comparative example and Example 1 is that tea leaves are not added, as detailed below: S1. Grinding and drying: Grind 5000g of shelled red pine nuts, put them in a 50℃ oven overnight to dry, and pass them through a 10-mesh sieve to obtain red pine nut powder; S2. Extraction of Korean pine kernel oil: 20g of pectinase, 40g of cellulase, 20g of protease and 20g of amylase were dissolved in 900g of water to obtain a compound enzyme aqueous solution. This solution was then added to 2000g of Korean pine nut powder and stirred. The mixture was then subjected to a 40℃ water bath and ultrasonic treatment at 300W for 120 minutes. The mixture was then placed in a high-pressure bag, sealed, and placed in a high-pressure device for high-pressure treatment at 25MPa for 5 minutes. After the treatment, the mixture was filtered and centrifuged to collect the Korean pine kernel oil. Comparative Example 2

[0026] The difference between this comparative example and Example 1 is that no complex enzyme is added, as detailed below: S1. Grinding and drying: Mix 5000g of shelled red pine nuts with 50g of black tea and 50g of green tea, grind them together, put them in a 50℃ oven to dry overnight, and pass them through a 10-mesh sieve to obtain mixed red pine nut powder; S2. Extraction of red pine kernel oil: Add 900g of water to 2000g of mixed red pine nut powder and stir. Soak in a 40℃ water bath and sonicate at 300W for 120 minutes. Put the mixture into a high-pressure bag, seal it, and place it in a high-pressure device for 25MPa high-pressure treatment for 5 minutes. After the treatment, filter and centrifuge to collect the red pine kernel oil. Comparative Example 3

[0027] The difference between this comparative example and Example 1 is that high-pressure treatment is not performed; only enzymatic extraction is used, as detailed below: S1. Grinding and drying: Mix 5000g of shelled red pine nuts with 50g of black tea and 50g of green tea, grind them together, put them in a 50℃ oven to dry overnight, and pass them through a 10-mesh sieve to obtain mixed red pine nut powder; S2. Extraction of Korean pine kernel oil: Dissolve 20g of pectinase, 40g of cellulase, 20g of protease and 20g of amylase in 900g of water to obtain a compound enzyme solution. Add 2000g of mixed Korean pine nut powder and stir. Soak in a 40℃ water bath and sonicate at 300W for 120min. After the mixture is removed, filter and centrifuge to collect the Korean pine kernel oil. Indicator Test

[0028] Oil yield measurement Oil yield (%) = (Oil weight / Raw material weight) × 100% The results are shown in Table 1. The oil yield of Example 1 was 90.8%. The oil yield of Comparative Example 1 was not significantly affected by the absence of tea leaves. The oil yield of Comparative Example 2, which did not undergo enzymatic hydrolysis, was 73.9%, and that of Comparative Example 3 was only 65.0%. This indicates that the combined use of compound enzymatic hydrolysis and high-pressure extraction can improve the oil yield and reduce losses.

[0029] Table 1. Oil yield and peroxide value of pine kernel oil extracted by different extraction methods

[0030] The method for testing peroxide value shall be performed in accordance with GB / T5538-2005 / ISO390.

[0031] The results are shown in Table 1. Example 1 had the lowest peroxide value, while Comparative Example 1 had the highest peroxide value, indicating that tea has a certain protective effect against the oxidation of oils. Example 2

[0032] A method for preparing a direct beverage made from red pine kernel oil that is beneficial to cardiovascular health includes the following steps: S1. Enzymatic hydrolysis: 0.6g papain and 0.6g bromelain were added to 198.8g PBS buffer to obtain a complex enzyme buffer. 20g whey protein and 20g casein were mixed with 100g complex enzyme buffer respectively. The pH was adjusted to 6, and the mixture was sonicated at 150W for 30min in a 35℃ water bath. The pH was adjusted to 5 again, and the mixture was sonicated at 500W for 30min to inactivate the enzyme. After inactivation, the pH was restored to 6 to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S2. Hawthorn pretreatment: Take 500g of washed and pitted hawthorn, cook until soft, crush and add 50g of 10wt.% sugar solution and boil again. Pass the solution through a 6-mesh sieve while hot to obtain hawthorn pulp. After a short cooling, place it in a 37℃ water bath. S3. Preparation of probiotic microparticles: 50g of Bifidobacterium suspension (1×10¹¹ CFU / g) and 50g of Saccharomyces boulardii suspension (1×10¹¹ CFU / g) were mixed to obtain a compound probiotic suspension. Then, it was mixed with 100g of 2wt.% lecithin solution at 150 rpm and stirred for 15min. 100g of enzymatically hydrolyzed whey protein solution and 10g of triglyceride hydrolase were added. After hydration at 37℃ and 300 rpm for 60min, it was added to 100g of hawthorn pulp, homogenized at 5MPa, and spray-dried to obtain probiotic microparticles. S4. Ingredient mixing: Mix 30g of probiotic microparticles, 100g of enzymatically hydrolyzed casein solution and 50g of fruit juice, then add to 600g of red pine kernel oil prepared in Example 1. Homogenize at 8000 rpm for 10 minutes and then perform microfluidic homogenization at 15 MPa to obtain red pine kernel oil direct beverage. Example 3

[0033] A method for preparing a direct beverage made from red pine kernel oil that is beneficial to cardiovascular health includes the following steps: S1. Proteolytic activity: 1.2g papain and 1.2g bromelain were added to 397.6g of PBS buffer to obtain a complex enzyme buffer. 40g of whey protein and 40g of casein were mixed with 200g of complex enzyme buffer, respectively. The pH was adjusted to 6, and the mixture was sonicated at 150W for 30min in a 35℃ water bath. The pH was adjusted to 5 again, and the mixture was sonicated at 500W for 30min to inactivate the enzyme. After inactivation, the pH was restored to 6 to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S2. Hawthorn pretreatment: Take 500g of washed and pitted hawthorn, cook until soft, crush and add 50g of 10wt.% sugar solution and boil again. Pass the solution through a 6-mesh sieve while hot to obtain hawthorn pulp. After a short cooling, place it in a 37℃ water bath. S3. Preparation of probiotic microparticles: 50g of Bifidobacterium suspension (1×10¹¹ CFU / g) and 50g of Saccharomyces boulardii suspension (1×10¹¹ CFU / g) were mixed to obtain a compound probiotic suspension. Then, it was mixed with 100g of 2wt.% lecithin solution at 150 rpm and stirred for 15min. 200g of enzymatically hydrolyzed whey protein solution and 10g of triglyceride hydrolase were added. After hydration at 37℃ and 300 rpm for 60min, it was added to 100g of hawthorn pulp, homogenized at 5MPa, and spray-dried to obtain probiotic microparticles. S4. Ingredient mixing: Mix 30g of probiotic microparticles, 100g of enzymatically hydrolyzed casein solution and 50g of fruit juice, then add to 600g of red pine kernel oil prepared in Example 1. Homogenize at 8000 rpm for 10 minutes and then perform microfluidic homogenization at 15 MPa to obtain red pine kernel oil direct beverage. Example 4

[0034] A method for preparing a direct beverage made from red pine kernel oil that is beneficial to cardiovascular health includes the following steps: S1. Enzymatic hydrolysis: 0.6g papain and 0.6g bromelain were added to 198.8g PBS buffer to obtain a complex enzyme buffer. 20g whey protein and 20g casein were mixed with 100g complex enzyme buffer respectively. The pH was adjusted to 6, and the mixture was sonicated at 150W for 30min in a 35℃ water bath. The pH was adjusted to 5 again, and the mixture was sonicated at 500W for 30min to inactivate the enzyme. After inactivation, the pH was restored to 6 to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S2. Hawthorn pretreatment: Take 500g of washed and pitted hawthorn, cook until soft, crush and add 50g of 10wt.% sugar solution and boil again. Pass the solution through a 6-mesh sieve while hot to obtain hawthorn pulp. After a short cooling, place it in a 37℃ water bath. S3. Preparation of probiotic microparticles: 50g of Bifidobacterium suspension (1×10¹¹ CFU / g) and 50g of Saccharomyces boulardii suspension (1×10¹¹ CFU / g) were mixed to obtain a compound probiotic suspension. Then, it was mixed with 100g of 2wt.% lecithin solution at 150 rpm and stirred for 15min. 100g of enzymatically hydrolyzed whey protein solution and 10g of triglyceride hydrolase were added. After hydration at 37℃ and 300 rpm for 60min, it was added to 200g of hawthorn pulp, homogenized at 5MPa, and spray-dried to obtain probiotic microparticles. S4. Ingredient mixing: Mix 30g of probiotic microparticles, 100g of enzymatically hydrolyzed casein solution and 50g of fruit juice, then add to 600g of red pine kernel oil prepared in Example 1. Homogenize at 8000 rpm for 10 minutes and then perform microfluidic homogenization at 15 MPa to obtain red pine kernel oil direct beverage. Example 5

[0035] A method for preparing a direct beverage made from red pine kernel oil that is beneficial to cardiovascular health includes the following steps: S1. Enzymatic hydrolysis: 0.6g papain and 0.6g bromelain were added to 198.8g PBS buffer to obtain a complex enzyme buffer. 20g whey protein and 20g casein were mixed with 100g complex enzyme buffer respectively. The pH was adjusted to 6, and the mixture was sonicated at 150W for 30min in a 35℃ water bath. The pH was adjusted to 5 again, and the mixture was sonicated at 500W for 30min to inactivate the enzyme. After inactivation, the pH was restored to 6 to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S2. Hawthorn pretreatment: Take 500g of washed and pitted hawthorn, cook until soft, crush and add 50g of 10wt.% sugar solution and boil again. Pass the solution through a 6-mesh sieve while hot to obtain hawthorn pulp. After a short cooling, place it in a 37℃ water bath. S3. Preparation of probiotic microparticles: 50g of Bifidobacterium suspension (1×10¹¹ CFU / g) and 50g of Saccharomyces boulardii suspension (1×10¹¹ CFU / g) were mixed to obtain a compound probiotic suspension. Then, it was mixed with 100g of 2wt.% lecithin solution at 150 rpm and stirred for 15min. 100g of enzymatically hydrolyzed whey protein solution and 10g of triglyceride hydrolase were added. After hydration at 37℃ and 300 rpm for 60min, it was added to 100g of hawthorn pulp, homogenized at 5MPa, and spray-dried to obtain probiotic microparticles. S4. Ingredient mixing: Mix 10g of probiotic microparticles, 100g of enzymatically hydrolyzed casein solution and 50g of fruit juice, then add to 600g of red pine kernel oil prepared in Example 1. Homogenize at 8000 rpm for 10 minutes and then perform microfluidic homogenization at 15 MPa to obtain red pine kernel oil direct beverage. Comparative Example 4

[0036] The difference between this comparative example and Example 3 is that the protein is not enzymatically digested, as detailed below: S1. Protein solution preparation: 40g of whey protein and 40g of casein were mixed with 200g of PBS buffer to obtain whey protein solution and casein solution, respectively. S2. Hawthorn pretreatment: Take 500g of washed and pitted hawthorn, cook until soft, crush and add 50g of 10wt.% sugar solution and boil again. Pass the solution through a 6-mesh sieve while hot to obtain hawthorn pulp. After a short cooling, place it in a 37℃ water bath. S3. Preparation of probiotic microparticles: 50g of Bifidobacterium suspension (1×10¹¹ CFU / g) and 50g of Saccharomyces boulardii suspension (1×10¹¹ CFU / g) were mixed to obtain a compound probiotic suspension, which was then mixed with 100g of 2wt.% lecithin solution at 150 rpm for 15 min. 200g of whey protein solution and 10g of triglyceride hydrolase were added, and the mixture was hydrated by stirring at 37℃ and 300 rpm for 60 min. After that, it was added to 100g of hawthorn pulp, homogenized at 5 MPa, and spray-dried to obtain probiotic microparticles. S4. Ingredient mixing: Mix 30g of probiotic microparticles, 100g of casein solution and 50g of fruit juice, then add to 600g of red pine kernel oil prepared in Example 1. Homogenize at 8000 rpm for 10 minutes and then perform microfluidic homogenization at 15 MPa to obtain red pine kernel oil direct beverage. Comparative Example 5

[0037] The difference between this comparative example and Example 3 is that probiotic microparticles are not prepared, as detailed below: S1. Proteolytic activity: 1.2g papain and 1.2g bromelain were added to 397.6g of PBS buffer to obtain a complex enzyme buffer. 40g of whey protein and 40g of casein were mixed with 200g of complex enzyme buffer, respectively. The pH was adjusted to 6, and the mixture was sonicated at 150W for 30min in a 35℃ water bath. The pH was adjusted to 5 again, and the mixture was sonicated at 500W for 30min to inactivate the enzyme. After inactivation, the pH was restored to 6 to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S2. Hawthorn pretreatment: Take 500g of washed and pitted hawthorn, cook until soft, crush and add 50g of 10wt.% sugar solution and boil again. Pass the solution through a 6-mesh sieve while hot to obtain hawthorn pulp. After a short cooling, place it in a 37℃ water bath. S4. Ingredient Mixing: 10g of Bifidobacterium suspension (1×10¹¹ CFU / g), 10g of Saccharomyces boulardii suspension (1×10¹¹ CFU / g), 1g of lecithin, 20g of whey protein solution, 10g of hawthorn pulp, 100g of enzymatically hydrolyzed casein solution and 50g of fruit juice were mixed and added to 600g of Korean pine kernel oil prepared in Example 1. The mixture was homogenized at 8000 rpm for 10 min and then homogenized at 15 MPa using microfluidic jet to obtain a direct Korean pine kernel oil beverage. Comparative Example 6

[0038] The difference between this comparative example and Example 3 is that probiotics are not added, as detailed below: S1. Proteolytic activity: 1.2g papain and 1.2g bromelain were added to 397.6g of PBS buffer to obtain a complex enzyme buffer. 40g of whey protein and 40g of casein were mixed with 200g of complex enzyme buffer, respectively. The pH was adjusted to 6, and the mixture was sonicated at 150W for 30min in a 35℃ water bath. The pH was adjusted to 5 again, and the mixture was sonicated at 500W for 30min to inactivate the enzyme. After inactivation, the pH was restored to 6 to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S2. Hawthorn pretreatment: Take 500g of washed and pitted hawthorn, cook until soft, crush and add 50g of 10wt.% sugar solution and boil again. Pass the solution through a 6-mesh sieve while hot to obtain hawthorn pulp. After a short cooling, place it in a 37℃ water bath. S3. Preparation of microparticles: 100g of 2wt.% lecithin solution was added to 200g of enzymatically hydrolyzed whey protein solution and 10g of triglyceride hydrolase. After stirring and hydrating at 37℃ and 300 rpm for 60min, it was added to 100g of hawthorn pulp, homogenized at 5MPa, and then spray-dried to obtain probiotic microparticles. S4. Ingredient mixing: Mix 30g microparticles, 100g enzymatically hydrolyzed casein solution and 50g fruit juice, then add to 600g of red pine kernel oil prepared in Example 1. Homogenize at 8000 rpm for 10 min and then perform microfluidic homogenization at 15 MPa to obtain red pine kernel oil direct beverage. Comparative Example 7

[0039] The difference between this comparative example and Example 3 is that hawthorn juice is not added, as detailed below: S1. Proteolytic activity: 1.2g papain and 1.2g bromelain were added to 397.6g of PBS buffer to obtain a complex enzyme buffer. 40g of whey protein and 40g of casein were mixed with 200g of complex enzyme buffer, respectively. The pH was adjusted to 6, and the mixture was sonicated at 150W for 30min in a 35℃ water bath. The pH was adjusted to 5 again, and the mixture was sonicated at 500W for 30min to inactivate the enzyme. After inactivation, the pH was restored to 6 to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S3. Preparation of probiotic microparticles: 50g of Bifidobacterium suspension (1×10¹¹ CFU / g) and 50g of Saccharomyces boulardii suspension (1×10¹¹ CFU / g) were mixed to obtain a compound probiotic suspension, which was then mixed with 100g of 2wt.% lecithin solution at 150 rpm for 15 min. 200g of enzymatically hydrolyzed whey protein solution and 10g of triglyceride hydrolase were added. After stirring and hydrating at 37℃ and 300 rpm for 60 min, the mixture was spray-dried to obtain probiotic microparticles. S4. Ingredient mixing: Mix 30g of probiotic microparticles, 100g of enzymatically hydrolyzed casein solution and 50g of fruit juice, then add to 600g of red pine kernel oil prepared in Example 1. Homogenize at 8000 rpm for 10 minutes and then perform microfluidic homogenization at 15 MPa to obtain red pine kernel oil direct beverage. Comparative Example 8

[0040] The difference between this comparative example and Example 3 is that fruit juice is not added, as detailed below: S1. Proteolytic activity: 1.2g papain and 1.2g bromelain were added to 397.6g of PBS buffer to obtain a complex enzyme buffer. 40g of whey protein and 40g of casein were mixed with 200g of complex enzyme buffer, respectively. The pH was adjusted to 6, and the mixture was sonicated at 150W for 30min in a 35℃ water bath. The pH was adjusted to 5 again, and the mixture was sonicated at 500W for 30min to inactivate the enzyme. After inactivation, the pH was restored to 6 to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S2. Hawthorn pretreatment: Take 500g of washed and pitted hawthorn, cook until soft, crush and add 50g of 10wt.% sugar solution and boil again. Pass the solution through a 6-mesh sieve while hot to obtain hawthorn pulp. After a short cooling, place it in a 37℃ water bath. S3. Preparation of probiotic microparticles: 50g of Bifidobacterium suspension (1×10¹¹ CFU / g) and 50g of Saccharomyces boulardii suspension (1×10¹¹ CFU / g) were mixed to obtain a compound probiotic suspension. Then, it was mixed with 100g of 2wt.% lecithin solution at 150 rpm and stirred for 15min. 200g of enzymatically hydrolyzed whey protein solution and 10g of triglyceride hydrolase were added. After hydration at 37℃ and 300 rpm for 60min, it was added to 100g of hawthorn pulp, homogenized at 5MPa, and spray-dried to obtain probiotic microparticles. S4. Ingredient mixing: Mix 30g of probiotic microparticles and 100g of enzymatically hydrolyzed casein solution, then add to 600g of red pine kernel oil prepared in Example 1. Homogenize at 8000 rpm for 10 minutes and then perform microfluidic homogenization at 15 MPa to obtain red pine kernel oil direct beverage. Performance testing

[0041] 1. Antioxidant property testing Malondialdehyde (MDA) testing mainly refers to GB / T5009.181-2003.

[0042] DPPH free radical scavenging rate determination Prepare 1.75×10 with ethanol -4 Take 2 mL of mol / L DPPH solution, dilute it to a certain concentration, dissolve the analyte in the DPPH solution, mix thoroughly, let stand for 30 min, and measure its absorbance at 517 nm.

[0043] In the formula: A i The absorbance is measured by comparing 2 mL of DPPH solution with 2 mL of sample solution. A j The absorbance is the result of mixing 2 mL of sample extract with 2 mL of ethanol. A c The absorbance is the result of mixing 2 mL of DPPH solution with 2 mL of ethanol.

[0044] Determination of ABTS free radical scavenging rate Take 1 mL of sample solution diluted to a certain concentration, add 3 mL of ABTS+ solution (7 mM ABTS solution and 2.45 mM K2S2O2 are mixed in equal proportions and reacted overnight (12 h-16 h) to prepare ABTS·+ stock solution. Adjust the absorbance of the ABTS·+ reaction solution at 734 nm to 0.70 ± 0.02), shake for 30 s, react at room temperature in the dark for 60 min, and then measure the absorbance value at 734 nm.

[0045] ABTS + Clearance rate (%) = (Acontrol - A test ) ∗100 / A contro In the formula: A control A is the absorbance value of the control tube. test This represents the absorbance of the sample to be tested.

[0046] The test results are shown in Table 2. Compared with Example 2, Example 3 showed that increasing the enzyme dosage released more antioxidant peptides, reduced malondialdehyde (MDA), and improved free radical scavenging rate. Hawthorn is rich in polyphenolic antioxidants, and increasing its dosage (Example 4) enhanced the antioxidant effect; removing hawthorn (Comparative Example 7) significantly reduced the scavenging rate. Fruit juice also has antioxidant effects; in Comparative Example 8, removing fruit juice increased oxidation products and decreased free radical scavenging ability. Probiotic microparticles can protect activity and enhance antioxidant stability; Comparative Example 5 did not prepare microparticles and had the highest MDA content.

[0047] Table 2. Antioxidant properties of different red pine kernel oil direct beverages

[0048] Sensory evaluation Using a comprehensive scoring method, a judging panel of 10 people (who meet the standards for food sensory analysis scorers) comprehensively examines the appearance and internal quality of the samples and assigns scores for each item and a total score.

[0049] As shown in Table 3, the hawthorn pulp dosage in Example 4 was doubled, resulting in a slight improvement in color and taste compared to other examples, as well as a slightly better appearance. Comparative Example 7, which did not add hawthorn, had the lowest color score, and its flavor was also affected. Comparative Example 5, which did not add microparticles, had a poorer taste, and its appearance was also affected by its decreased emulsification stability. Overall, the sensory scores of the examples were all above 90 points, while the comparative examples were all lower than the examples.

[0050] Table 3 Sensory evaluation scores of different red pine kernel oil direct beverages Stability evaluation The emulsion release index is an indicator of stability. The lower the emulsion release index, the higher the emulsion stability. Refer to the method for determining the emulsion release index to measure stability.

[0051]

[0052] in, H c The height of the upper level H t This is the total height.

[0053] The physical stability of the products in the examples was higher than that in the comparative examples. In comparative example 4, the protein was not enzymatically hydrolyzed, resulting in poor emulsification and significantly reduced stability. In comparative example 7, no hawthorn pulp was added, and the microparticles were prepared without a carrier, leading to poor stability. In comparative example 8, the absence of fruit juice had a relatively small impact on stability; the lack of microparticles had a significant impact. Comparative example 5 did not prepare microparticles, making it prone to stratification. Although comparative example 6 did not contain probiotics, the presence of microparticles preserved some of the emulsification properties, resulting in a lower stratification index.

[0054] Table 4. Stratification Index of Different Red Pine Seed Oil Direct Drinks Functional testing The effects of ICR mice on blood lipids were investigated. The high-fat diet consisted of 500g basal feed, 90g lard, 9g cholesterol, 0.9g bile salts, 0.9g propylthiouracil, and 300mL water. After one week of acclimatization, the ICR mice were divided into four groups: a normal control group, a high-fat model group, an example group (containing pine kernel oil), and a control group (containing pine kernel oil). Each group consisted of 10 mice, and the experiment lasted 28 days. Except for the normal control group, which was fed the basal feed, all other groups were fed the same amount of high-fat feed, with free access to food and water. Simultaneously, the example and control groups of the pine kernel oil were also administered the same amount of sample via gavage. After the acclimatization period, the mice were fasted for 12 hours. Blood was collected from the orbital sinus the following day, and serum was separated to determine triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C).

[0055] The results in Table 5 show that the high-fat diet model was successfully constructed because the differences in various indicators between it and the normal control group were significant. Compared with the high-fat diet model group, the serum TC and LDL-C levels and TG levels in the sample group were significantly reduced, while the HDL-C level was significantly increased, indicating that both Example 3 and the comparative example had lipid-lowering effects. The bioactive peptides produced by enzymatic hydrolysis can inhibit cholesterol synthesis and promote lipid metabolism. Without enzymatic hydrolysis (Comparative Example 4), TG, TC, and LDL-C levels were significantly increased, while HDL-C was decreased. Hawthorn acid in hawthorn can inhibit cholesterol absorption, and flavonoids can regulate lipid metabolism. After removal (Comparative Example 7), the LDL-C clearance capacity decreased.

[0056] Table 5. Lipid-lowering ability of different red pine kernel oil direct drinks

[0057] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A method for preparing a direct beverage made from red pine kernel oil that is beneficial to cardiovascular health, characterized in that, Includes the following steps: S1. Grinding and Drying: The shelled red pine nuts are mixed with tea leaves and ground together. They are then placed in an oven to dry overnight and sieved to obtain mixed red pine nut powder. S2. Extraction of Korean pine kernel oil: Add the compound enzyme aqueous solution to the mixed Korean pine nut powder and stir. Sonicate in a water bath, pack into a high-pressure bag, seal and place in a high-pressure device for high-pressure treatment. After the treatment, filter and centrifuge to collect the Korean pine kernel oil. S3. Enzymatic hydrolysis: Whey protein and casein are added to a complex enzyme buffer, pH is adjusted, and the mixture is sonicated in a water bath. The pH and sonication power are adjusted again to inactivate the enzyme. After inactivation, the pH is restored to obtain the enzymatically hydrolyzed whey protein solution and casein solution. S4. Hawthorn pretreatment: Wash the hawthorns, remove the pits, cook them, crush them, add sugar solution again and boil them, sieve them while hot to obtain hawthorn pulp, cool them briefly and place them in a water bath; S5. Preparation of probiotic microparticles: Mix and stir the compound probiotic suspension and lecithin solution, add the enzymatically hydrolyzed whey protein solution and lipase, stir and hydrate, add to hawthorn pulp, homogenize and spray dry to obtain probiotic microparticles; S6. Ingredient Mixing: After mixing probiotic microparticles, enzymatically hydrolyzed casein solution and fruit juice, add to red pine kernel oil, homogenize at high speed and then perform microfluidic homogenization to obtain red pine kernel oil direct beverage.

2. The method for preparing a direct-drink beverage of red pine kernel oil that is beneficial to cardiovascular health according to claim 1, characterized in that, In step S1, the mass ratio of red pine nuts to tea leaves is (50-100):1; the temperature of the drying oven is 50-60℃; and the sieve used for sieving is 10-20 mesh.

3. The method for preparing a direct-drink beverage of red pine kernel oil that is beneficial to cardiovascular health according to claim 2, characterized in that, The tea leaves are a mixture of green tea and black tea.

4. The method for preparing a direct-drink beverage of red pine kernel oil that is beneficial to cardiovascular health according to claim 1, characterized in that, In step S2, the compound enzyme is a mixture of pectinase, cellulase, protease, and amylase in a mass ratio of (1-2):(2-3):(1-2):1; the concentration of the compound enzyme aqueous solution is 10-20 wt.%; the mass ratio of the compound enzyme aqueous solution to the mixed red pine nut powder is (3-5):10; the temperature of the water bath ultrasound is 35-45℃, the time of the water bath ultrasound is 90-180 min, and the power of the water bath ultrasound is 200-500 W; the pressure of the high-pressure treatment is 25-45 MPa, and the time of the high-pressure treatment is 5-10 min.

5. The method for preparing a direct-drink beverage of red pine kernel oil that is beneficial to cardiovascular health according to claim 1, characterized in that, In step S3, the complex enzyme is papain and bromelain in a mass ratio of 1:1, and the enzyme content in the complex enzyme buffer is 0.6-1 wt.%; the mass ratio of whey protein, casein and complex enzyme buffer is (1-2):10; adjust the pH to 6-8; the water bath temperature is 35-40℃, the ultrasonic power is 100-200W, and the ultrasonic time is 30-60min; adjust the pH again to 5-6, and the ultrasonic power is 500-600W; restore the pH to 6-8.

6. The method for preparing a direct-drink beverage of red pine kernel oil that is beneficial to cardiovascular health according to claim 1, characterized in that, In step S4, the mass ratio of hawthorn to sugar solution is (5-10):1, the concentration of sugar solution is 10-15 wt.%, the sieve used is 6 mesh, and the water bath temperature is 30-37℃.

7. The method for preparing a direct-drink beverage of red pine kernel oil that is beneficial to cardiovascular health according to claim 1, characterized in that, In step S5, the compound probiotics consist of Bifidobacterium and Saccharomyces boulardii in a colony count ratio of (1-2):1, with a concentration of 10¹¹ CFU / g; the concentration of the lecithin solution is 1-2 wt.%; the mass ratio of the compound probiotic suspension, lecithin solution, enzymatically hydrolyzed whey protein solution, lipase, and hawthorn pulp is 10:10:(10-20):1:(10-20); the stirring speed is 100-200 rpm, and the stirring time is 10-15 min; the stirring speed for hydration is 200-300 rpm, and the stirring time is 30-60 min, with a temperature of 37℃; the homogenization pressure is 5-10 MPa.

8. The method for preparing a direct-drink beverage of red pine kernel oil that is beneficial to cardiovascular health according to claim 7, characterized in that, The lipase is a triglyceride hydrolase.

9. The method for preparing a direct-drink beverage of red pine kernel oil that is beneficial to cardiovascular health according to claim 1, characterized in that, In step S6, the fruit juice is one or a combination of apple and pomegranate juice; the mass ratio of probiotic microparticles, enzymatically hydrolyzed casein solution, fruit juice and red pine kernel oil is (1-3):10:(3-6):(50-70); the high-speed homogenization speed is 8000-10000 rpm, the high-speed homogenization time is 10-15 min; the microjet homogenization pressure is 15-20 MPa.

10. A red pine kernel oil direct beverage prepared according to any one of claims 1-9, which is beneficial to cardiovascular health.

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