A preparation method of fucoxanthin microcapsules

Abstract

The invention discloses a method for preparing fucoxanthin microcapsules, which belongs to the technical field of food and drug processing, and comprises the following steps: fucoxanthin, carrageenan and protein complex are mixed, and after stirring reaction, fucoxanthin lipid nanoparticles are obtained; microcrystalline cellulose is grafted and modified with succinic anhydride, and then mixed with food-grade chromium picolinate and fucoxanthin lipid nanoparticles to obtain composite fucoxanthin; composite fucoxanthin, propolis extract, white kidney bean extract, pomegranate peel extract, quercetin and tea polyphenols are mixed as core materials, and β-cyclodextrin and gum arabic are used as capsule materials to prepare microcapsules. Fucoxanthin is present in lipid nanoparticles, and the nano size can better adhere to the cell mitochondria to stimulate the mitochondrial effect; the formed microcapsules contain network structures and lipid nanoparticles inside, which can exert a strong force on the composite fucoxanthin and avoid leakage of fucoxanthin in the microcapsules.

Description

Technical Field

[0001] The invention relates to the technical field of food and medicine processing, and in particular to a method for preparing fucoxanthin microcapsules. Background Art

[0002] Fucoxanthin, also known as fucoxanthin and brown adipose tissue, is an important and highly active carotenoid. Through light-proof extraction, the content of fucoxanthin is only about one hundred thousandth, and it is known as the rare algae gold. Fucoxanthin can stimulate mitochondrial metabolic function, improve the overall cell metabolism level, especially promote the transformation of fat-storing white fat cells into fat-burning brown fat cells, thereby significantly reducing the weight of white adipose tissue (WAT) in the waist and abdomen; secondly, it can regulate the expression of mitochondrial uncoupling protein 1 (UCPI), promote fat decomposition and heat production conversion, reduce blood, liver and subcutaneous fat, thereby reflecting the overall healthy fat-reducing and body-shaping effect, inhibiting fat production, reducing fat deposition, balancing energy intake, improving leptin resistance, and enhancing fat oxidation, avoiding obesity-induced chronic diseases.

[0003] Fucoxanthin can burn fat, eliminate fat accumulation, and reduce white adipose tissue in the waist and abdomen, reflecting the overall healthy fat reduction and static fat reduction effects, but fucoxanthin is unstable to light, heat, oxygen, acid and alkali, and is easily oxidized and isomerized, which limits its application as a nutritional supplement in the food industry; encapsulating fucoxanthin to prepare microcapsules can effectively improve the stability of fucoxanthin, but existing microcapsules easily expel the encapsulated fucoxanthin, causing fucoxanthin leakage. Summary of the invention

[0004] The object of the present invention is to provide a method for preparing fucoxanthin microcapsules: fucoxanthin, carrageenan and a protein complex are mixed and stirred for reaction; the protein complex zein and whey protein isolate have amphiphilicity, so that the hydrophobic fucoxanthin is dispersed in the carrageenan and protein mixture to form lipid nanoparticles; the fucoxanthin exists in the lipid nanoparticles, thereby improving the thermal stability of the fucoxanthin; its nano size can better adhere to the cell mitochondria, thereby stimulating the mitochondrial effect and enhancing the static fat-reducing effect; succinic anhydride grafted modified microcrystalline cellulose is then mixed with food-grade chromium picolinate and fucoxanthin lipid nanoparticles; the anhydride-modified microcrystalline cellulose and the fucoxanthin lipid nanoparticles are The particles and food-grade chromium picolinate form a hydrogen-bonded cross-linked network nanoemulsion, which further enhances the effect on the fucoxanthin lipid nanoparticles and avoids the leakage of the fucoxanthin lipid nanoparticles; the composite fucoxanthin, propolis extract, white kidney bean extract, pomegranate peel extract, quercetin, and tea polyphenols are mixed as the core material, and β-cyclodextrin and gum arabic are used as capsule materials to achieve β-cyclodextrin and gum arabic wrapping of the composite, thereby improving the stability of the composite, and the formed microcapsules contain a network structure and lipid nanoparticles inside, which can have a stronger effect on the composite fucoxanthin and avoid the leakage of fucoxanthin in the microcapsules; after the microcapsule shell material is decomposed, the released lipid nanoparticles can better exert the fat-reducing effect.

[0005] The technical problem to be solved by the present invention is that fucoxanthin is unstable to light, heat, oxygen, acid and alkali, and is easily oxidized and isomerized, which limits its application as a nutritional supplement in the food industry; encapsulating fucoxanthin to prepare microcapsules can effectively improve the stability of fucoxanthin, but the existing microcapsules are prone to expel the encapsulated fucoxanthin, causing the fucoxanthin to leak.

[0006] To achieve the above object, the present invention provides the following technical solutions:

[0007] A method for preparing fucoxanthin microcapsules comprises the following steps:

[0008] S1. Mix fucoxanthin, carrageenan and protein complex, and stir to react to obtain fucoxanthin lipid nanoparticles.

[0009] Furthermore, the protein complex is prepared by mixing zein and whey protein isolate in a mass ratio of (2-3):(1-2).

[0010] Furthermore, the particle size of the fucoxanthin lipid nanoparticles is 50-80 nm.

[0011] Furthermore, the fucoxanthin lipid nanoparticles are prepared by the following steps:

[0012] The protein complex was added to an ethanol aqueous solution with a mass fraction of 85%, and stirred evenly to obtain a composite solution. Carrageenan was added to 50 mL of ethanol, and stirred evenly. The composite solution and fucoxanthin were added and stirred evenly. 1 mL of apple cider vinegar with a concentration of 0.2 mol / L was added to adjust the pH value to 4. The mixture was stirred at 700 r / min for 1 h, and the ethanol was removed by rotary evaporation at 40°C and -0.1 MPa. Deionized water was added, and the mixture was centrifuged at 500 r / min for 30 min to remove large particles. The mixture was filtered, washed with deionized water, and dried in an oven at 4°C overnight to obtain fucoxanthin lipid nanoparticles.

[0013] Among them, carrageenan is rich in sulfate groups and is negatively charged. It can electrostatically adsorb the positive charges carried by zein and whey protein isolate to improve the stability of the protein complex, thereby enhancing the antioxidant properties of zein and whey protein isolate, and further improving the antioxidant properties of fucoxanthin.

[0014] Furthermore, zein and whey protein isolate are amphiphilic, so that the hydrophobic fucoxanthin is dispersed in the carrageenan and protein mixture to form lipid nanoparticles, so that the fucoxanthin exists in the lipid nanoparticles, thereby improving the thermal stability of the fucoxanthin. The nanosize effect of the lipid nanoparticles can better adhere to the cell mitochondria, thereby stimulating the mitochondrial effect and enhancing the static fat-reducing effect.

[0015] Furthermore, the ratio of the protein complex to the ethanol aqueous solution is (2-3) g (15-25) mL.

[0016] Furthermore, the dosage ratio of carrageenan, ethanol, composite solution, fucoxanthin, and apple cider vinegar is (1.2-1.6) g: (45-55) mL: (3-5) mL: (1-2) g: (0.5-1.5) mL.

[0017] S2. Succinic anhydride is used to graft-modify microcrystalline cellulose, and then mixed with food-grade chromium picolinate and fucoxanthin lipid nanoparticles to obtain composite fucoxanthin; specifically prepared by the following steps:

[0018] Succinic anhydride grafted modified microcrystalline cellulose and food-grade chromium picolinate were added to deionized water, heated to 50°C, stirred evenly, and fucoxanthin lipid nanoparticles were added. After stirring at 30°C for 4 hours, a nanoemulsion, namely composite fucoxanthin, was formed.

[0019] Among them, the anhydride-treated microcrystalline cellulose can complex with the chromium ions in food-grade chromium picolinate to form a hydrogen bond cross-linked network, and the carboxyl groups contained in the anhydride-treated microcrystalline cellulose can combine with the hydrophilic groups contained on the surface of the fucoxanthin lipid nanoparticles through chemical bonds, so that the fucoxanthin lipid nanoparticles are dispersed in the hydrogen bond cross-linked network, further enhancing the force of the fucoxanthin lipid nanoparticles and avoiding the leakage of the fucoxanthin lipid nanoparticles.

[0020] Furthermore, the dosage ratio of succinic anhydride grafted modified microcrystalline cellulose, food-grade chromium picolinate, and deionized water is (3-4) g: (1.5-1.9) mL: (45-55) mL.

[0021] Furthermore, the succinic anhydride grafted modified microcrystalline cellulose is prepared by the following steps:

[0022] Succinic anhydride is added to ethanol, stirred evenly to obtain anhydride solution, microcrystalline cellulose is added to 80% ethanol aqueous solution by mass fraction, stirred evenly, the anhydride solution is added, the temperature is raised to 35°C, a 2 mol / L sodium bicarbonate aqueous solution is added to adjust the pH to 8.5, the reaction is stirred for 2 hours to form a solid, the solid is filtered, washed 3 times with ethanol, washed 3 times with deionized water, and dried in an oven at 55°C overnight to obtain succinic anhydride grafted modified microcrystalline cellulose.

[0023] Among them, microcrystalline cellulose has good adhesion, which improves the adhesion of fucoxanthin to cells, stimulates the function of cell mitochondria, and enhances the fat-reducing effect. Sodium bicarbonate aqueous solution is used as a catalyst to enable the hydroxyl groups contained in microcrystalline cellulose to undergo esterification reaction with succinic anhydride, and introduce more oxygen-containing functional group carboxyl groups on microcrystalline cellulose, thereby improving the adsorption capacity of microcrystalline cellulose on fucoxanthin lipid nanoparticles and preventing the microcapsules from easily expelling the encapsulated fucoxanthin and causing the leakage of fucoxanthin.

[0024] Furthermore, the ratio of succinic anhydride to ethanol is (1-2) g:(8-12) mL.

[0025] Furthermore, the dosage ratio of microcrystalline cellulose, ethanol aqueous solution, acid anhydride solution, and sodium bicarbonate aqueous solution is (3-4) g: (70-90) mL: (13-17) mL: (0.2-0.3) g.

[0026] S3. Prepare microcapsules by mixing composite fucoxanthin, propolis extract, white kidney bean extract, pomegranate peel extract, quercetin and tea polyphenols as core materials, and using β-cyclodextrin and gum arabic as capsule materials. The microcapsules are prepared by the following steps:

[0027] Add composite fucoxanthin, propolis extract, white kidney bean extract, pomegranate peel extract, quercetin and tea polyphenols to 70% ethanol aqueous solution by mass, stir evenly at 2000 rpm to obtain core material, add β-cyclodextrin and gum arabic to ethanol and deionized water, stir evenly, add core material, heat to 60°C, stir and react at 1000 rpm for 5 hours for embedding, filter with a vacuum pump to obtain a solid, freeze-dry the solid at -15°C to obtain microcapsules.

[0028] Among them, the composite fucoxanthin, propolis extract, white kidney bean extract and pomegranate peel extract can be mixed with quercetin and tea polyphenols, and after high-speed stirring, the quercetin, tea polyphenols and extracts are evenly mixed. During the stirring process, the quercetin and tea polyphenols in the core material are hydrogen bonded to the composite fucoxanthin to form a stable cross-linked network structure, further increasing the stability of the composite fucoxanthin; β-cyclodextrin and gum arabic are used as capsule materials, the outer surface has free hydroxyl groups and is hydrophilic, while the internal cavity is basically formed by glucose molecules connected by glycosidic bonds, so it is hydrophobic, thereby achieving the encapsulation of the complex by β-cyclodextrin and gum arabic.

[0029] Furthermore, the formed microcapsules have a network structure inside, which can exert a stronger force on the composite fucoxanthin and avoid leakage of the fucoxanthin in the microcapsules.

[0030] Furthermore, the mass ratio of composite fucoxanthin, propolis extract, white kidney bean extract, pomegranate peel extract, quercetin, and tea polyphenols is (1-2) g:(0.2-0.4):(0.1-0.3):(0.1-0.2):(0.05-0.15):(0.2-0.3).

[0031] Furthermore, the mass ratio of β-cyclodextrin to gum arabic is (2.5-2.7):(1.3-1.5).

[0032] Further, the propolis extract is specifically prepared by the following steps:

[0033] The crude propolis was ground in a -20°C refrigerated grinder, taken out, added to an 80% by mass ethanol aqueous solution, stirred for 24 hours, hexane was added to remove wax and non-polar substances, the filtrate was collected by filtration, the filtrate was treated in a 40°C rotary evaporator for 1 hour to remove the solvent, and freeze-dried at -10°C for 10 minutes to obtain a propolis extract.

[0034] Furthermore, the usage ratio of crude propolis, ethanol aqueous solution and hexane is (8-12) g:(90-110) mL:(4-6) g.

[0035] Furthermore, the white kidney bean extract is specifically prepared by the following steps:

[0036] The white kidney bean was soaked in deionized water at 4°C for 12 hours and peeled, taken out, placed in an oven at 35°C for 10 minutes, crushed with a grinder, and passed through a 100-mesh sieve to obtain white kidney bean powder. The white kidney bean powder was added to deionized water, stirred at 50°C for 2.5 hours, 0.5 mol / L hydrochloric acid was added to adjust the pH to 3, and the centrifuge was collected at 10000 r / min for 20 minutes to obtain the white kidney bean extract.

[0037] Furthermore, the dosage ratio of white kidney bean powder, deionized water and hydrochloric acid is (4-6) g: (90-110) mL: (0.3-0.7) mL.

[0038] Furthermore, the pomegranate peel extract is specifically prepared by the following steps:

[0039] The pomegranate peel was ground with a grinder, added to 70% ethanol, stirred evenly, placed in a microwave oven with a power of 264 W, and microwave extracted for 1.5 minutes. The extract was taken out, the filtrate was collected by filtration, concentrated with a rotary evaporator, and freeze-dried in a -20°C freeze dryer to obtain a pomegranate peel extract.

[0040] Furthermore, the ratio of pomegranate peel to ethanol is (2.1-2.5) g: (40-50) mL.

[0041] Furthermore, compared with the prior art, the present invention has the following beneficial effects:

[0042] (1) In the technical scheme of the present invention, carrageenan, zein and whey protein isolate are electrostatically adsorbed to improve the stability of the protein complex, thereby enhancing the antioxidant properties of zein and whey protein isolate, and further improving the antioxidant properties of fucoxanthin; zein and whey protein isolate are amphiphilic, so that hydrophobic fucoxanthin is dispersed in the carrageenan and protein mixture to form lipid nanoparticles. On the one hand, fucoxanthin exists in the lipid nanoparticles, which improves the thermal stability of fucoxanthin. Its nano size can better adhere to the cell mitochondria, thereby stimulating the mitochondrial effect and enhancing the static fat-reducing effect. On the other hand, carrageenan has good stability under acid-base conditions, thereby improving the stability of fucoxanthin under acid-base conditions. In addition, whey protein isolate can promote fat burning and help reduce fat. During the fat-reducing process, whey protein isolate contains rich essential amino acids, which helps muscle growth and repair, and has the effect of reducing fat and increasing muscle; zein can reduce the generation of free radicals in the body, has significant antioxidant properties, and protects cells from oxidative damage during fat reduction.

[0043] (2) In the technical solution of the present invention, succinic anhydride can promote the energy metabolism of human cells, enhance the metabolic activity of myocardial cells, and improve human immunity and antioxidant capacity; microcrystalline cellulose is rich in dietary fiber, which can promote gastrointestinal motility and the secretion of digestive juices. To a certain extent, it can also achieve the effect of promoting digestion and reducing fat, and has good adhesion, thereby improving the adhesion of fucoxanthin to cells, stimulating the function of cell mitochondria, and enhancing the fat-reducing effect. In addition, microcrystalline cellulose can replace part of the medium-chain triglycerides in fat, increase the consumption of calories, and accelerate fat metabolism.

[0044] (3) In the technical scheme of the present invention, microcrystalline cellulose and succinic anhydride undergo an esterification reaction, and more oxygen-containing functional group carboxyl groups are introduced into the microcrystalline cellulose, thereby improving the adsorption capacity of microcrystalline cellulose to fucoxanthin lipid nanoparticles, preventing the microcapsules from easily expelling the encapsulated fucoxanthin, causing the leakage of fucoxanthin. The anhydride-treated microcrystalline cellulose, fucoxanthin lipid nanoparticles, and food-grade chromium picolinate form a hydrogen-bonded cross-linked network nanoemulsion, which further enhances the force on the fucoxanthin lipid nanoparticles and avoids the leakage of fucoxanthin lipid nanoparticles. In addition, as a chromium dietary supplement, food-grade chromium picolinate has a relatively small content and will not affect the human body. It improves insulin sensitivity and glucose metabolism, reduces body fat, and makes insulin more sensitive by increasing insulin activity or up-regulating insulin receptors, thereby helping to control body weight.

[0045] (4) In the technical solution of the present invention, the propolis extract contains a large amount of polyphenol structures, which has an anti-obesity effect. It can reduce weight by improving lipid metabolism and reducing insulin resistance. The flavonoids and non-flavonoids contained in it can effectively fight free radicals that cause oxidative stress, have good antioxidant properties, and can enhance the antioxidant properties of fucoxanthin; it can also prevent lipid peroxidation, platelet aggregation and capillary permeability. White kidney bean extract contains a protein-based α-amylase inhibitor, which acts as a "starch blocker" by binding to the target enzyme α-amylase, inhibiting amylase, reducing carbohydrate digestion, reducing blood sugar content in postprandial plasma, controlling fat accumulation, and having a good anti-obesity function. The flavonoids and polyphenols found in white kidney bean extract and propolis extract have a synergistic effect in reducing hyperlipidemia and avoiding lipid peroxidation.

[0046] (5) In the technical scheme of the present invention, the pomegranate peel extract can effectively remove free radicals and exhibit an antioxidant effect. The lipase inhibitor contained in the pomegranate peel extract reduces the synthesis of fatty acids in the human body, increases energy consumption, and achieves the purpose of fat reduction. The ellagic acid contained in the pomegranate peel extract has an inhibitory effect on human preadipocytes, and achieves the purpose of fat reduction by inhibiting the differentiation of human preadipocytes and proliferating them into mature fat cells. The pomegranate peel extract contains an α-amylase inhibitor propolis extract, which has a synergistic effect in anti-obesity. Quercetin can effectively reduce myocardial hypoxia damage, promote the generation of endogenous energy, improve mitochondrial function after myocardial ischemia, increase the activity of mitochondrial complex enzymes, stimulate mitochondria in cells, and promote the generation of adenosine triphosphate, the energy required by cells; tea polyphenols can effectively reduce weight by activating the sympathetic nervous system, releasing norepinephrine, stimulating peripheral fat oxidation, and increasing heat production in the body. It can also inhibit the activity of α-glucosidase by regulating enzyme activity, thereby inhibiting the increase in blood sugar and reducing the conversion of carbohydrates to fat, thereby preventing obesity and controlling weight. In addition, tea polyphenols have a high inhibitory activity on pancreatic lipase, which can reduce the body's absorption of triglycerides in food, inhibit the formation and accumulation of fat, and have a good effect in anti-obesity.

[0047] (6) In the technical scheme of the present invention, propolis extract, white kidney bean extract, pomegranate peel extract, quercetin, tea polyphenols and composite fucoxanthin are combined through hydrogen bonds to form a stable core material, β-cyclodextrin and gum arabic are used as capsule materials, and the composite is wrapped by β-cyclodextrin and gum arabic, thereby improving the stability of the composite. The formed microcapsules contain a network structure and lipid nanoparticles inside, which can exert a strong force on the composite fucoxanthin and avoid leakage of fucoxanthin in the microcapsules. After the microcapsule shell material is decomposed, the released lipid nanoparticles can better exert the fat-reducing effect. DETAILED DESCRIPTION

[0048] The technical solutions in the embodiments of the present invention are described clearly and completely. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0049] The raw materials used in the examples of the present invention are as follows: fucoxanthin (purity 98%, purchased from Chengdu Desite Biotechnology Co., Ltd.); quercetin (batch number BD105676, purity 99%, purchased from Henan Aoke Standard Material Technology Co., Ltd.); tea polyphenols (purity 95%, purchased from Xi'an Dongfeng Biotechnology Co., Ltd.); carrageenan (batch number: GF2022071101A, purchased from Fujian Lvxin Food Co., Ltd.); zein (AR, amylose content 26%, molecular weight 168.29kD a); whey protein isolate (purchased from Shanghai Maclean Biochemical Technology Co., Ltd.); chromium picolinate (food grade, purity 99%, purchased from Shandong Fuhe Biotechnology Co., Ltd.); succinic anhydride (analytical grade, Shanghai Maclean Biochemical Technology Co., Ltd.); microcrystalline cellulose (Tesco Chemical (Hubei) Co., Ltd., purity 99%); propolis (Zhejiang Jiangshan Hengliang Bee Products Co., Ltd.); β-cyclodextrin (analytical grade, Henan Wanbang Industrial Co., Ltd.); gum arabic (food grade, Tianjin Fengchuan Chemical Reagent Technology Co., Ltd.).

[0050] Example 1

[0051] Fucoxanthin lipid nanoparticles were prepared by the following steps:

[0052] A1. Mix 2.5 g of zein and 1.5 g of whey protein isolate to obtain a protein complex.

[0053] A2. Add 2.5 g of the protein complex to 20 mL of 85% ethanol-water solution, stir well to obtain a composite solution, add 1.4 g of carrageenan to 50 mL of ethanol, stir well, add 4 mL of the composite solution and 1.5 g of fucoxanthin, stir well, add 1 mL of 0.2 mol / L apple cider vinegar to adjust the pH value to 4, stir at 700 r / min for 1 h, remove ethanol by rotary evaporation at 40°C and -0.1 MPa, add 50 mL of deionized water, centrifuge at 500 r / min for 30 min to remove large particles, filter, wash with deionized water, and dry in an oven at 4°C overnight to obtain fucoxanthin lipid nanoparticles.

[0054] Comparative Example 1

[0055] The difference between this comparative example and Example 1 is that carrageenan is not added, and the remaining steps and raw materials are the same as those of Example 1.

[0056] A1. Mix 2.5 g of zein and 1.5 g of whey protein isolate to obtain a protein complex.

[0057] A2. Add 2.5 g of protein complex to 20 mL of 85% ethanol-water solution, stir evenly, add 1.5 g of fucoxanthin and 1 mL of 0.2 mol / L apple cider vinegar to adjust the pH to 4, stir at 700 r / min for 1 h, remove ethanol by rotary evaporation at 40°C and -0.1 MPa, add 50 mL of deionized water, centrifuge at 500 r / min for 30 min to remove large particles, filter, wash with deionized water, and dry in an oven at 4°C overnight to obtain fucoxanthin lipid nanoparticles.

[0058] Example 2

[0059] The composite fucoxanthin is specifically prepared by the following steps:

[0060] B1. Add 1.5 g of succinic anhydride to 10 mL of ethanol and stir well to obtain anhydride solution. Add 3.5 g of microcrystalline cellulose to 80 mL of 80% ethanol aqueous solution and stir well. Add 15 mL of anhydride solution, heat to 35°C, add 2 mol / L sodium bicarbonate aqueous solution to adjust the pH to 8.5, stir and react for 2 hours to form a solid. The solid is filtered, washed with ethanol 3 times, washed with deionized water 3 times, and dried in an oven at 55°C overnight to obtain succinic anhydride grafted modified microcrystalline cellulose.

[0061] B2. Add 3.5 g of succinic anhydride grafted modified microcrystalline cellulose and 1.7 g of food-grade chromium picolinate into 50 mL of deionized water, heat to 50°C, stir evenly, add 1.5 g of fucoxanthin lipid nanoparticles, and stir at 30°C for 4 hours to form a nanoemulsion, i.e., composite fucoxanthin.

[0062] Comparative Example 2

[0063] The difference between this comparative example and Example 2 is that the fucoxanthin lipid nanoparticles are replaced with the substance prepared in Comparative Example 1, and the remaining steps and raw materials are the same as those in Example 2.

[0064] Comparative Example 3

[0065] The difference between this comparative example and Example 2 is that the succinic anhydride grafted modified microcrystalline cellulose is replaced by microcrystalline cellulose, and the remaining steps are synchronized with Example 2.

[0066] 3.5 g of modified microcrystalline cellulose and 1.7 g of food-grade chromium picolinate were added to 50 mL of deionized water, heated to 50°C, stirred evenly, 1.5 g of fucoxanthin lipid nanoparticles were added, and stirred at 30°C for 4 h to form a nanoemulsion, namely, composite fucoxanthin.

[0067] Comparative Example 4

[0068] The difference between this comparative example and Example 2 is that succinic anhydride grafted modified microcrystalline cellulose and food-grade chromium picolinate are not added, and the remaining steps are synchronized with Example 2.

[0069] Mix 3.6 g of 1.2 mol / L calcium hydroxide aqueous solution and 2.5 g of 0.6 mol / L phosphoric acid aqueous solution, stir evenly, add 2.6 g of diatomaceous earth, mix at 400 rpm for 30 min, heat to 60° C., stir to react for 2 h, cool to room temperature, filter, wash 3 times with deionized water, dry in an oven at 65° C. for 10 min, place in 30 mL of ethanol, add 10 g of urea, stir evenly, stir at 85° C. until the organic solvent evaporates, cool to room temperature, and obtain a loaded urea complexing agent.

[0070] Comparative Example 5

[0071] The difference between this comparative example and Example 2 is that the fucoxanthin lipid nanoparticles are replaced with fucoxanthin, and the remaining steps are synchronized with Example 2.

[0072] B1. Add 1.5 g of succinic anhydride to 10 mL of ethanol and stir well to obtain anhydride solution. Add 3.5 g of microcrystalline cellulose to 80 mL of 80% ethanol aqueous solution and stir well. Add 15 mL of anhydride solution, heat to 35°C, add 2 mol / L sodium bicarbonate aqueous solution to adjust the pH to 8.5, stir and react for 2 hours to form a solid. The solid is filtered, washed with ethanol 3 times, washed with deionized water 3 times, and dried in an oven at 55°C overnight to obtain succinic anhydride grafted modified microcrystalline cellulose.

[0073] B2. Add 3.5 g of succinic anhydride grafted modified microcrystalline cellulose and 1.7 g of food-grade chromium picolinate to 50 mL of deionized water, heat to 50°C, stir evenly, add 1.5 g of fucoxanthin, and stir at 30°C for 4 hours to form a nanoemulsion, i.e., composite fucoxanthin.

[0074] Example 3

[0075] Propolis extract is specifically prepared by the following steps:

[0076] Grind 10 g of crude propolis in a -20°C refrigerated grinder, take it out, add it to 100 mL of 80% ethanol aqueous solution, stir it for 24 hours, add 5 g of hexane to remove wax and non-polar substances, filter and collect the filtrate, treat the filtrate in a rotary evaporator at 40°C for 1 hour to remove the solvent, and freeze-dry it at -10°C for 10 minutes to obtain propolis extract.

[0077] Example 4

[0078] The white kidney bean extract is specifically prepared by the following steps:

[0079] Soak white kidney beans in deionized water at 4°C for 12 hours and peel them, take them out, place them in an oven at 35°C to dry for 10 minutes, grind them with a grinder, and pass them through a 100-mesh sieve to obtain white kidney bean powder. Take 5g of white kidney bean powder and add it to 100mL of deionized water, stir it at 50°C for 2.5 hours, add 0.5mL of 0.5mol / L hydrochloric acid to adjust the pH to 3, centrifuge it at 10000r / min for 20 minutes, collect the centrifuge, and obtain the white kidney bean extract.

[0080] Example 5

[0081] The pomegranate peel extract is specifically prepared by the following steps:

[0082] Grind 2.3 g of pomegranate peel with a grinder, add it to 46 mL of 70% ethanol, stir evenly, place it in a microwave oven with a power of 264 W, perform microwave extraction for 1.5 min, take it out, collect the filtrate by filtration, concentrate it with a rotary evaporator, and freeze-dry it in a -20°C freeze dryer to obtain a pomegranate peel extract.

[0083] Example 6

[0084] A method for preparing fucoxanthin microcapsules comprises the following steps:

[0085] 1g of composite fucoxanthin, 0.2g of propolis extract, 0.1g of white kidney bean extract, 0.1g of pomegranate peel extract, 0.05g of quercetin and 0.2g of tea polyphenols were added to 80mL of 70% ethanol aqueous solution, stirred at 2000rpm to obtain core material, 2.5g of β-cyclodextrin and 1.3g of gum arabic were added to 10mL of ethanol and 20mL of deionized water, stirred evenly, 10g of core material was added, the temperature was raised to 60°C, and the reaction was stirred at 1000rpm for 5h for embedding, and the solid was obtained by filtration with a vacuum pump. The solid was freeze-dried at -15°C to obtain microcapsules.

[0086] Example 7

[0087] A method for preparing fucoxanthin microcapsules comprises the following steps:

[0088] 1.5g of composite fucoxanthin, 0.3g of propolis extract, 0.2g of white kidney bean extract, 0.15g of pomegranate peel extract, 0.1g of quercetin and 0.25g of tea polyphenols were added to 80mL of 70% ethanol aqueous solution, stirred at 2000rpm to obtain core material, 2.6g of β-cyclodextrin and 1.4g of gum arabic were added to 10mL of ethanol and 20mL of deionized water, stirred evenly, 10g of core material was added, the temperature was raised to 60°C, and the reaction was stirred at 1000rpm for 5h for embedding, and the solid was obtained by filtration with a vacuum pump. The solid was freeze-dried at -15°C to obtain microcapsules.

[0089] Example 8

[0090] A method for preparing fucoxanthin microcapsules comprises the following steps:

[0091] 2g of composite fucoxanthin, 0.4g of propolis extract, 0.3g of white kidney bean extract, 0.2g of pomegranate peel extract, 0.15g of quercetin and 0.3g of tea polyphenols were added to 80mL of 70% ethanol aqueous solution, stirred at 2000rpm to obtain core material, 2.7g of β-cyclodextrin and 1.5g of gum arabic were added to 10mL of ethanol and 20mL of deionized water, stirred evenly, 10g of core material was added, the temperature was raised to 60°C, and the reaction was stirred at 1000rpm for 5h for embedding, and the solid was obtained by filtration with a vacuum pump. The solid was freeze-dried at -15°C to obtain microcapsules.

[0092] Comparative Example 6

[0093] The difference between this comparative example and Example 7 is that the composite fucoxanthin is replaced by the substance prepared in Comparative Example 2, and the remaining steps and raw materials are the same as those in Example 7.

[0094] Comparative Example 7

[0095] The difference between this comparative example and Example 7 is that the composite fucoxanthin is replaced by the substance prepared in Comparative Example 3, and the remaining steps are synchronized with Example 7.

[0096] Comparative Example 8

[0097] The difference between this comparative example and Example 7 is that the composite fucoxanthin is replaced by the substance prepared in Comparative Example 4, and the remaining steps are synchronized with Example 7.

[0098] Comparative Example 8

[0099] The difference between this comparative example and Example 7 is that the composite fucoxanthin is replaced by the substance prepared in Comparative Example 5, and the remaining steps are synchronized with Example 7.

[0100] Comparative Example 9

[0101] The difference between this comparative example and Example 7 is that propolis extract, white kidney bean extract, pomegranate peel extract, quercetin and tea polyphenols are not added, and the remaining steps are synchronized with Example 7.

[0102] The microcapsules prepared in Examples 6-8 and Comparative Examples 6-9 were tested.

[0103] Fat reduction performance test: select people aged 25-50 years old, who are overweight, do not like to exercise, have a high body fat rate or constipation problems, and those who need to reduce fat, with a body weight of 62.5kg and a body fat rate of 27.14%; take the above-prepared microcapsules for four consecutive weeks, twice a day in the morning and evening, 2 mg each time; measure the average body weight (kg) after taking it for 2 weeks and 4 weeks, and calculate the average body fat rate after taking it for 1 week and 2 weeks;

[0104] Stability test: Determination of retention rate of fucoxanthin microcapsules: Take a certain mass of fucoxanthin microcapsules and place them in a stoppered test tube, heat them with air blast at 60°C for 8 days, take samples respectively, and determine the retention rate, and perform quantitative analysis by liquid chromatography. The retention rate is calculated according to the following formula: Fucoxanthin retention rate (%) = fucoxanthin microcapsule content after heating / initial fucoxanthin microcapsule content × 100%.

[0105] Determination of embedding rate: the embedding rate prepared above was placed in a 10mL centrifuge tube, 5ml petroleum ether (boiling range 30-60°C) was added, shaken and centrifuged for 10min, the supernatant was taken, diluted with anhydrous ethanol, and the peak area was measured at 460nm by high performance liquid chromatography (HPLC), and the amount of fucoxanthin on the surface of the microcapsule was calculated; the fucoxanthin microcapsules were added to 10mL phosphate buffer with a pH of 7, neutral protease was added, the wall was broken by shaking, suction filtered, and the filter paper was washed with anhydrous ethanol, the filtrate was collected and rotary evaporated to dryness, the rotary evaporator was washed with an appropriate amount of anhydrous ethanol, the eluent was diluted with anhydrous ethanol to a certain concentration, and the total content of fucoxanthin in the microcapsule was measured by HPLC; the embedding rate of the microcapsule was calculated in this way, and repeated three times, and the calculation formula is as follows: Fucoxanthin microcapsule embedding rate % = (1-free fucoxanthin content on the microcapsule surface / total fucoxanthin content in the microcapsule × 100%); the test results are shown in Table 1 below:

[0106] Table 1

[0107]

[0108] It can be seen from the data in Table 1 that in Comparative Example 5, the fucoxanthin lipid nanoparticles prepared without adding carrageenan are used to prepare microcapsules, and their stability and antioxidant properties are reduced. This may be because carrageenan improves the antioxidant properties of fucoxanthin through electrostatic adsorption with zein and whey protein isolate, and carrageenan has good stability under acid and alkali conditions, thereby improving the stability of fucoxanthin under acid and alkali conditions; Comparative Example 6 replaces succinic anhydride grafted modified microcrystalline cellulose with microcrystalline cellulose for preparing microcapsules, and its fat-reducing effect and microcapsule embedding rate are reduced. This may be because more oxygen-containing functional group carboxyl groups are introduced into microcrystalline cellulose to improve the adsorption of microcrystalline cellulose on fucoxanthin lipid nanoparticles, avoid microcapsules from easily expelling the encapsulated fucoxanthin, and succinic anhydride can promote the energy metabolism of human cells, enhance the metabolic activity of myocardial cells, and improve human immunity and antioxidant capacity; Comparative Example 7 does not add succinic anhydride grafted modified microcrystalline cellulose and food-grade chromium picolinate to prepare microcapsules with composite fucoxanthin The stability and embedding rate of the capsule decreased, which may be because the anhydride microcrystalline cellulose and the food-grade chromium picolinate formed a hydrogen-bonded cross-linked network nanoemulsion, which enhanced the force on the fucoxanthin lipid nanoparticles and avoided the leakage of the fucoxanthin lipid nanoparticles. In addition, the food-grade chromium picolinate, as a chromium dietary supplement, can reduce body fat. In Comparative Example 8, the fucoxanthin lipid nanoparticles were replaced with fucoxanthin to prepare the composite fucoxanthin microcapsules, and the fat-reducing function thereof decreased. This may be because the fucoxanthin exists in the lipid nanoparticles, which improves the thermal stability of the fucoxanthin, and its nano size can better adhere to the cell mitochondria, thereby stimulating the mitochondrial effect and enhancing the static fat-reducing effect. In Comparative Example 9, the microcapsules prepared without the addition of propolis extract, white kidney bean extract, pomegranate peel extract, quercetin, and tea polyphenols had a decreased fat-reducing function and fucoxanthin stability compared with the examples. The addition of propolis extract, white kidney bean extract, pomegranate peel extract, quercetin, and tea polyphenols was beneficial to improving the stability and fat-reducing performance of the microcapsules.

[0109] The data in Table 1 show that the fucoxanthin microcapsules prepared in Examples 6-8 not only have good fat-reducing efficacy and stability, but also have good embedding rate and better mitochondrial stimulation. Fucoxanthin, carrageenan and protein complex are mixed and stirred to react to obtain fucoxanthin lipid nanoparticles; microcrystalline cellulose is grafted and modified with succinic anhydride, and then mixed with food-grade chromium picolinate and fucoxanthin lipid nanoparticles to obtain composite fucoxanthin; composite fucoxanthin, propolis extract, white kidney bean extract, pomegranate peel extract, quercetin and tea polyphenols are mixed as core materials, β-cyclodextrin and gum arabic are used as capsule materials, and microcapsules are prepared to meet the test performance requirements, while the microcapsules prepared in Comparative Examples 5-9 do not meet the performance requirements, indicating that the fucoxanthin microcapsules prepared in the present invention not only have good fat-reducing efficacy and stability, but also have good embedding rate and better mitochondrial stimulation.

[0110] In the description of the specification, the description with reference to the terms "one embodiment", "example", "specific example", etc. means that the specific features, structures, materials or characteristics described in conjunction with the embodiment or example are included in at least one embodiment or example of the present invention. In this specification, the schematic representation of the above terms does not necessarily refer to the same embodiment or example. Moreover, the specific features, structures, materials or characteristics described can be combined in any one or more embodiments or examples in a suitable manner.

[0111] The above contents are merely examples and explanations of the present invention. Those skilled in the art may make various modifications or additions to the specific embodiments described or replace them in a similar manner. As long as they do not deviate from the invention or exceed the scope defined by the claims, they shall all fall within the protection scope of the present invention.

Claims

1. A method for preparing fucoxanthin microcapsules, characterized in that: Including the following raw materials by weight: S1. mixing fucoxanthin, carrageenan and protein complex, stirring and reacting to obtain fucoxanthin lipid nanoparticles; The protein complex is formed by mixing zein and whey protein isolate in a mass ratio of (2-3):(1-2); The mass ratio of the fucoxanthin, carrageenan and protein complex is (1.5-1.7):(0.5-0.7):(2-3); The particle size of the fucoxanthin lipid nanoparticles is 50-80 nm; S2. Succinic anhydride is used to graft and modify microcrystalline cellulose, and then mixed with food-grade chromium picolinate and fucoxanthin lipid nanoparticles to obtain composite fucoxanthin; The succinic anhydride grafted modified microcrystalline cellulose is specifically prepared by the following steps: The succinic anhydride is added to ethanol and stirred evenly to obtain an anhydride solution, the microcrystalline cellulose is added to an ethanol aqueous solution with a mass fraction of 80%, stirred evenly, the anhydride solution is added, the temperature is raised to 35°C, a 2 mol / L sodium bicarbonate aqueous solution is added to adjust the pH to 8.5, the reaction is stirred for 2 hours to form a solid, and the solid is filtered, washed, and dried to obtain the succinic anhydride grafted modified microcrystalline cellulose; The ratio of succinic anhydride to ethanol is (1-2) g: (8-12) mL; The amount ratio of the microcrystalline cellulose, ethanol aqueous solution, acid anhydride solution, and sodium bicarbonate aqueous solution is (3-4) g: (70-90) mL: (13-17) mL: (0.2-0.3) g; S3. Prepare microcapsules by mixing composite fucoxanthin, propolis extract, white kidney bean extract, pomegranate peel extract, quercetin and tea polyphenols as core materials, and using β-cyclodextrin and gum arabic as capsule materials.

2. The method for preparing fucoxanthin microcapsules according to claim 1, characterized in that: The propolis extract is specifically prepared by the following steps: The crude propolis was ground in a -20°C refrigerated grinder, taken out, added to an 80% by mass ethanol aqueous solution, stirred for 24 hours, hexane was added to remove wax and non-polar substances, the filtrate was collected by filtration, the filtrate was treated in a rotary evaporator at 40°C for 1 hour, and freeze-dried at -10°C for 10 minutes to obtain a propolis extract.

3. The method for preparing fucoxanthin microcapsules according to claim 1, characterized in that: The white kidney bean extract is specifically prepared by the following steps: The white kidney bean was soaked in deionized water at 4°C for 12 hours and peeled, taken out, placed in an oven at 35°C for 10 minutes, crushed with a grinder, and passed through a 100-mesh sieve to obtain white kidney bean powder. The white kidney bean powder was added to deionized water, stirred at 50°C for 2.5 hours, 0.5 mol / L hydrochloric acid was added to adjust the pH to 3, and the centrifuge was collected at 10000 r / min for 20 minutes to obtain the white kidney bean extract.

4. The method for preparing fucoxanthin microcapsules according to claim 1, characterized in that: The pomegranate peel extract is specifically prepared by the following steps: The pomegranate peel was ground with a grinder, added to 70% ethanol, stirred evenly, placed in a microwave oven with a power of 264 W, and microwave extracted for 1.5 minutes. The extract was taken out, the filtrate was collected by filtration, concentrated with a rotary evaporator, and freeze-dried in a -20°C freeze dryer to obtain a pomegranate peel extract.

5. The method for preparing fucoxanthin microcapsules according to claim 1, characterized in that: The mass ratio of the β-cyclodextrin to gum arabic is (2.5-2.7):(1.3-1.5).

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