Lycopene capsule as well as preparation method and application thereof in enhancing immunity

Abstract

The invention provides a lycopene capsule as well as a preparation method and application thereof in enhancing immunity, and relates to the technical field of health food. The lycopene capsule comprises the following raw materials: lycopene powder, pyrroloquinoline quinone disodium salt and a compound stabilizer, wherein the compound stabilizer is a mixture of an emblic leafflower fruit extract, an echinacea purpurea extract and D-sodium erythorbate. The lycopene capsule prepared by the invention has the effects of being high in stability and remarkably improving the immunity.

Description

Technical Field

[0001] The present invention relates to the technical field of health-care foods, and in particular to a lycopene capsule, a preparation method thereof and an application thereof in enhancing immunity. Background Art

[0002] Immunity is the body's ability to resist infection by pathogens. The human immune system has three protective functions: ① Defense: Resisting invasion by pathogenic microorganisms such as bacteria, viruses, and fungi; ② Stabilization: Promptly removing aging and dead cells from the body to prevent them from interfering with normal body functions, thereby maintaining internal stability; and ③ Immune Surveillance: Promptly detecting and eliminating mutated cells in the body to prevent the onset of cancer. The human immune system plays a crucial role in enabling humans to survive healthily in various environments. When the body is affected by a host of factors, such as adverse environmental conditions, diet, and emotional states, individual cells can mutate. Under normal circumstances, the immune system eliminates these mutated cells. However, in individuals with weakened immunity, their immune cells are unable to recognize these mutated cells, allowing them to grow and multiply, ultimately leading to cancer. Therefore, strengthening immunity and enhancing the body's resistance to pathogens has become a growing health priority.

[0003] Lycopene is an important carotenoid with a unique long-chain molecular structure and more unsaturated double bonds than other carotenoids. This unique structure provides excellent free radical scavenging and antioxidant capabilities, with its antioxidant capacity being twice that of beta-carotene and 100 times that of vitamin E. Lycopene's health benefits include delaying aging, preventing cancer, enhancing immunity, improving beauty, and protecting against ultraviolet radiation. It has become a new research hotspot in the international health food, cosmetics, and pharmaceutical sectors.

[0004] Chinese patent application CN103520361A discloses a lycopene compound preparation that enhances immunity and protects against chronic diseases. A composition comprising lycopene oleoresin, selenium-enriched yeast, and arginine as active ingredients, along with pharmaceutically acceptable excipients, can be prepared into soft capsules. The lycopene oleoresin contains lycopene and safflower oil. The composition of this invention can stimulate the body's own disease resistance, enhance the body's immune and digestive systems, and prevent and treat chronic diseases and sub-health conditions. Chinese patent application CN105638931A discloses a composite natural health supplement that enhances immunity, comprising lycopene, ganoderma lucidum extract, and edible oil. The nutritional supplement described in this invention can effectively improve the user's immunity. However, the technical solutions of the above-mentioned patent applications do not reflect testing of product stability.

[0005] Due to the 11 conjugated double bonds and 2 non-conjugated double bonds in the lycopene molecule, lycopene is relatively unstable and can undergo cis-trans isomerization and oxidative degradation under certain conditions. As a physiologically active substance, lycopene is susceptible to oxidation and decomposition under the influence of factors such as light, heat, oxygen, and pH, thereby losing its physiologically active functions and nutritional value. Furthermore, lycopene is a fat-soluble pigment, soluble in other lipids and non-polar solvents but insoluble in water. This instability and water insolubility significantly limit its application and promotion.

[0006] PQQ (pyrroloquinolinequinone) is a cofactor for various oxidoreductases and is widely found in plants and animals. Its powerful free radical scavenging function plays a vital role in the body. PQQ has been discovered to be the third redox cofactor in the redox cycle, following niacinamide and riboflavin. It is an antioxidant that participates in mitochondrial function, scavenges reactive oxygen species (ROS), and effectively protects human dermal fibroblasts from UVA-induced aging, improves cell damage, and inhibits collagen degradation.

[0007] However, there are currently few studies on the combined use of lycopene and PQQ. Based on this, it is necessary to provide a lycopene capsule with lycopene and PQQ as the main active ingredients, high stability, and the ability to significantly improve immunity. Summary of the Invention

[0008] In response to the above problems, the present invention provides a lycopene capsule, a preparation method and application thereof. By optimizing the components, ratios and preparation method, the obtained lycopene capsule is more stable, storage-resistant and has better immunity-enhancing activity.

[0009] To achieve the above object, the technical solution adopted by the present invention is as follows: In a first aspect, the present invention provides a lycopene capsule comprising the following raw materials: lycopene powder, pyrroloquinoline quinone disodium salt and a composite stabilizer, wherein the composite stabilizer is a mixture of emblica bud extract, echinacea purpurea extract and sodium D-isoascorbate.

[0010] Preferably, the mass ratio of the emblica extract, the echinacea extract and sodium D-isoascorbate is 10-20:1-10:1-3.

[0011] Preferably, the mass ratio of the emblica extract, the echinacea extract and sodium D-isoascorbate is 12-18:1-8:1-2.

[0012] Further preferably, the mass ratio of the emblica extract, the echinacea extract and sodium D-isoascorbate is 14:5:1.

[0013] Preferably, the preparation method of the emblica extract is: mixing emblica with water, heating and refluxing extraction 1-3 times to obtain a water extract and residue, gradient alcohol extraction of the residue to obtain an alcohol extract, mixing the water extract and the alcohol extract, and drying to obtain the emblica extract.

[0014] Preferably, the material-liquid ratio of the emblica buds to water is 1:8-15 g:L; Further preferably, the solid-liquid ratio of the emblica buds to water is 1:10 g:L.

[0015] Preferably, the step of gradient alcohol extraction is: the residue is mixed with 35wt%-45wt% ethanol solution for one alcohol extraction, then the concentration of the ethanol solution is increased to 50wt%-65wt% for a second alcohol extraction, and then the concentration of the ethanol solution is increased to 75wt%-85wt% for a third alcohol extraction, and the mixture is combined to obtain an alcohol extract.

[0016] Further preferably, the preparation method of the emblica extract is: the emblica is crushed, passed through a 40-60 mesh sieve, mixed with water at a solid-liquid ratio of 1:10 g:L, heated and refluxed for extraction 3 times to obtain a water extract and residue, the residue is mixed with a 40wt% ethanol solution for one alcohol extraction, and then the concentration of the ethanol solution is increased to 55wt% for a second alcohol extraction, and then the concentration of the ethanol solution is increased to 80wt% for a third alcohol extraction, combined to obtain an alcohol extract, the water extract and the alcohol extract are mixed, and spray dried to obtain the emblica extract.

[0017] Preferably, the preparation method of the Echinacea extract is: mixing Echinacea with alcohol, extracting with alcohol at 40-50° C. for 1-3 times, combining the alcohol extracts, and drying.

[0018] Preferably, the alcohol is a 75wt%-90wt% ethanol solution.

[0019] Preferably, the material-liquid ratio of Echinacea to alcohol is 1:8-12 g:L.

[0020] Further preferably, the preparation method of the Echinacea extract is: mixing Echinacea with 80wt% ethanol solution at a solid-liquid ratio of 1:10g:L, extracting with alcohol three times at 45°C, combining the alcohol extracts, and drying.

[0021] Preferably, the lycopene capsules comprise the following raw materials in parts by mass: 40-160 parts of lycopene powder, 1-10 parts of pyrroloquinoline quinone disodium salt and 1-12 parts of a composite stabilizer.

[0022] More preferably, the lycopene capsules comprise the following raw materials in parts by mass: 50-150 parts of lycopene powder, 2-10 parts of pyrroloquinoline quinone disodium salt and 1-10 parts of a composite stabilizer.

[0023] More preferably, the lycopene capsules comprise the following raw materials in parts by mass: 100 parts of lycopene powder, 10 parts of pyrroloquinoline quinone disodium salt and 8 parts of composite stabilizer.

[0024] Preferably, the lycopene capsules may further include auxiliary materials.

[0025] Preferably, the auxiliary materials include but are not limited to excipients, buffers, emulsifiers, diluents, binders, preservatives, lubricants, pH regulators, cryoprotectants, flavorings, fillers and antioxidants.

[0026] Preferably, the excipient is selected from at least one of microcrystalline cellulose, lactose, pregelatinized starch, cyclodextrin, carboxymethyl cellulose, mannitol, magnesium stearate, starch, calcium phosphate, ethyl cellulose, methyl cellulose, alginic acid, gelatin, gum arabic, glyceryl monostearate, sodium starch glycolate, guar gum, glycerol, and propylene glycol.

[0027] Preferably, the buffer is selected from at least one of sodium dihydrogen phosphate, sodium bicarbonate, ammonium bicarbonate, sodium acetate, citrate, histidine, and succinate.

[0028] Preferably, the emulsifier is selected from at least one of magnesium stearate, zinc stearate, calcium stearate, glyceryl stearate, sorbitan isostearate, sorbitan oleate, glyceryl oleate, and polyglyceryl-3 polyricinoleate.

[0029] Preferably, the diluent is selected from at least one of erythritol, mannitol, sorbitol, xylitol, lactose, sucrose, corn starch, potato starch, calcium phosphate, calcium citrate, and crystalline cellulose.

[0030] Preferably, the binder is selected from at least one of ethanol, starch slurry, pregelatinized starch, dextrin, syrup, hydroxypropyl methylcellulose, methylcellulose, sodium carboxymethylcellulose, ethyl cellulose, polyvinyl alcohol, polyethylene glycol, sodium alginate, polyvinyl pyrrolidone, gum arabic, gelatin, and alginic acid.

[0031] Preferably, the preservative is selected from at least one of methylparaben, propylparaben, methylparaben, ethylparaben, propylparaben, chlorobutanol, thimerosal, mercuric cyanide, phenoxyethanol, chlorhexidine, benzoic acid, sodium benzoate, chlorocresol, benzalkonium bromide, benzalkonium chloride, and ethylparaben.

[0032] Preferably, the lubricant is selected from at least one of magnesium stearate, zinc stearate, glyceryl monostearate, polyethylene glycol, stearic acid, talc, sodium chloride, sodium oleate, sodium lauryl sulfate, magnesium lauryl sulfate, sodium stearyl fumarate, poloxamer, and silicon dioxide.

[0033] Preferably, the pH adjuster is at least one selected from citric acid, fumaric acid, succinic acid, tartaric acid, malic acid and ascorbic acid.

[0034] Preferably, the flavoring agent is selected from at least one of sweet orange flavor, vanilla flavor, strawberry flavor, milk flavor, banana flavor, and cherry flavor.

[0035] Preferably, the filler is selected from at least one of mannitol, xylitol, sorbitol, maltose, microcrystalline cellulose, glucose, lactose, sucrose, dextrin, starch, sodium alginate, and sodium bicarbonate.

[0036] Preferably, the antioxidant can be selected from at least one of L-cysteine ​​hydrochloride, L-cysteine ​​base, 4,4 (2,3-dimethyltetramethylene dicatechol), tocopherol-rich extract (natural vitamin E), α-tocopherol (synthetic vitamin E), β-tocopherol, 6-tocopherol, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), propyl gallate, octyl gallate, dodecyl gallate, tert-butylhydroquinone (TBHQ), fumaric acid, malic acid, ascorbic acid (vitamin C), sodium ascorbate, calcium ascorbate, potassium ascorbate, ascorbyl palmitate, and ascorbyl stearate.

[0037] Preferably, the auxiliary material can be selected from at least one of microcrystalline cellulose, pregelatinized starch, magnesium stearate and silicon dioxide.

[0038] In a second aspect, the present invention provides a method for preparing the above-mentioned lycopene capsules, comprising the following steps: S1: Pyrroloquinoline quinone disodium salt is mixed with excipients to obtain mixture 1; S2: Lycopene powder is mixed with a composite stabilizer to obtain mixture 2; S3: Mix mixture 1 and mixture 2, and put them into empty capsules to obtain the product.

[0039] Preferably, the specification of the lycopene capsule is 0.3g / capsule.

[0040] In a third aspect, the present invention provides the use of the lycopene capsules described above in the preparation of health-care foods for improving immunity.

[0041] Compared with the prior art, the present invention has the following beneficial effects: 1. The present invention optimizes the components and uses lycopene powder and pyrroloquinoline quinone disodium salt (PQQ) as functional ingredients. Experiments have found that the two have a synergistic effect in improving immunity and strengthening the body.

[0042] 2. The present invention uses emblica extract, echinacea extract and sodium D-isoascorbate as composite stabilizers. Under the joint action of the three, the stability of the product is significantly improved, the storage time is greatly extended, and when used in conjunction with functional ingredients, the immunity-enhancing effect of the product is further improved.

[0043] 3. The specific components and proportions of the present invention can produce a product with high stability and significantly improve immunity. DETAILED DESCRIPTION

[0044] In order to make the technical means, creative features, purpose and effect of the present invention easy to understand, the present invention is further illustrated below in conjunction with specific embodiment, but the following embodiment is only a preferred embodiment of the present invention, not all. Based on the embodiment in the embodiment, other embodiments obtained by those skilled in the art without making creative work all fall within the protection scope of the present invention. It is worth noting that the raw materials used in the present invention are all common commercial products, and their source is not specifically limited. The technology and scientific terms used in the embodiment have the meaning commonly understood by those of ordinary skill in the art to which the present invention belongs.

[0045] Example 1 A lycopene capsule comprises the following raw materials in parts by mass: 100 parts of lycopene powder, 10 parts of pyrroloquinoline quinone disodium salt and 8 parts of composite stabilizer.

[0046] The composite stabilizer is a mixture of emblica bud extract, echinacea purpurea extract and sodium D-isoascorbate (mass ratio is 14:5:1).

[0047] The preparation method of the emblica fruit extract comprises the following steps: crushing the emblica fruit, passing through a 40-60 mesh sieve, mixing with water at a solid-liquid ratio of 1:10 g:L, heating and refluxing for extraction three times to obtain a water extract and residues, mixing the residues with a 40wt% ethanol solution for one alcohol extraction, then increasing the concentration of the ethanol solution to 55wt% for a second alcohol extraction, then increasing the concentration of the ethanol solution to 80wt% for a third alcohol extraction, combining the mixture to obtain an alcohol extract, mixing the water extract with the alcohol extract, spray drying, and obtaining the emblica fruit extract.

[0048] The preparation method of the echinacea extract is as follows: echinacea is mixed with 80wt% ethanol solution at a material-liquid ratio of 1:10g:L, the mixture is subjected to alcohol extraction at 45°C for 3 times, the alcohol extracts are combined, and the extracts are dried.

[0049] The preparation method of lycopene capsules is as follows: S1: Pyrroloquinoline quinone disodium salt is mixed with excipients such as microcrystalline cellulose, pregelatinized starch, magnesium stearate, and silicon dioxide, and the mixture is passed through a 40-60 mesh sieve to obtain a mixture 1; S2: Lycopene powder and composite stabilizer are mixed and passed through a 40-60 mesh sieve to obtain mixture 2; S3: Mix mixture 1 and mixture 2 evenly, and put them into empty capsules to obtain the product.

[0050] The specification of lycopene capsules is 0.3g / capsule.

[0051] Example 2 A lycopene capsule comprises the following raw materials in parts by mass: 40 parts of lycopene powder, 1 part of pyrroloquinoline quinone disodium salt and 1 part of composite stabilizer.

[0052] The composite stabilizer is a mixture of emblica oleracea extract, echinacea purpurea extract and sodium D-isoascorbate (mass ratio is 10:1:1).

[0053] The preparation method of the emblica extract comprises the following steps: crushing the emblica, passing through a 40-60 mesh sieve, mixing with water at a solid-liquid ratio of 1:8 g:L, heating and refluxing for extraction once to obtain a water extract and residues, mixing the residues with a 35wt% ethanol solution for alcohol extraction once, then increasing the concentration of the ethanol solution to 65wt% for alcohol extraction twice, then increasing the concentration of the ethanol solution to 75wt% for alcohol extraction three times, combining to obtain an alcohol extract, mixing the water extract with the alcohol extract, spray drying, and obtaining the emblica extract.

[0054] The preparation method of the echinacea extract is as follows: echinacea is mixed with a 90wt% ethanol solution at a material-liquid ratio of 1:8g:L, the mixture is subjected to alcohol extraction once at 40°C, the alcohol extracts are combined, and the extracts are dried.

[0055] The preparation method of lycopene capsules is as follows: S1: Pyrroloquinoline quinone disodium salt is mixed with excipients such as microcrystalline cellulose, pregelatinized starch, magnesium stearate, and silicon dioxide, and the mixture is passed through a 40-60 mesh sieve to obtain a mixture 1; S2: Lycopene powder and composite stabilizer are mixed and passed through a 40-60 mesh sieve to obtain mixture 2; S3: Mix mixture 1 and mixture 2 evenly, and put them into empty capsules to obtain the product.

[0056] The specification of lycopene capsules is 0.3g / capsule.

[0057] Example 3 A lycopene capsule comprises the following raw materials in parts by mass: 160 parts of lycopene powder, 9 parts of pyrroloquinoline quinone disodium salt and 12 parts of composite stabilizer.

[0058] The composite stabilizer is a mixture of emblica bud extract, echinacea purpurea extract and sodium D-isoascorbate (mass ratio is 20:10:3).

[0059] The preparation method of the emblica extract comprises the following steps: crushing the emblica, passing through a 40-60 mesh sieve, mixing with water at a solid-liquid ratio of 1:15 g:L, heating and refluxing for extraction three times to obtain a water extract and residues, mixing the residues with a 35wt% ethanol solution for one alcohol extraction, then increasing the concentration of the ethanol solution to 50wt% for a second alcohol extraction, then increasing the concentration of the ethanol solution to 85wt% for a third alcohol extraction, combining the mixture to obtain an alcohol extract, mixing the water extract with the alcohol extract, spray drying, and obtaining the emblica extract.

[0060] The preparation method of the Echinacea extract is as follows: Echinacea is mixed with a 75wt% ethanol solution at a material-liquid ratio of 1:12g:L, the mixture is subjected to alcohol extraction three times at 50°C, the alcohol extracts are combined, and the extracts are dried.

[0061] The preparation method of lycopene capsules is as follows: S1: Pyrroloquinoline quinone disodium salt is mixed with excipients such as microcrystalline cellulose, pregelatinized starch, magnesium stearate, and silicon dioxide, and the mixture is passed through a 40-60 mesh sieve to obtain a mixture 1; S2: Lycopene powder and composite stabilizer are mixed and passed through a 40-60 mesh sieve to obtain mixture 2; S3: Mix mixture 1 and mixture 2 evenly, and put them into empty capsules to obtain the product.

[0062] The specification of lycopene capsules is 0.3g / capsule.

[0063] Comparative Example 1 A lycopene capsule, compared with Example 1, uses lycopene powder instead of pyrroloquinoline quinone disodium salt, specifically as follows: Calculated by weight, it includes the following raw materials: 110 parts of lycopene powder and 8 parts of compound stabilizer.

[0064] The preparation method is as follows: S1: Microcrystalline cellulose, pregelatinized starch, magnesium stearate, and silicon dioxide are mixed and passed through a 40-60 mesh sieve to obtain mixture 1; S2: Lycopene powder and composite stabilizer are mixed and passed through a 40-60 mesh sieve to obtain mixture 2; S3: Mix mixture 1 and mixture 2 evenly, and put them into empty capsules to obtain the product.

[0065] The rest is the same as Example 1.

[0066] Comparative Example 2 A lycopene capsule, compared with Example 1, uses Ganoderma lucidum extract and camellia oil instead of pyrroloquinoline quinone disodium salt, specifically as follows: Calculated by weight, it includes the following raw materials: 100 parts of lycopene powder, 1.25 parts of ganoderma lucidum extract, 8.75 parts of camellia oil and 8 parts of compound stabilizer.

[0067] The preparation method is as follows: S1: Ganoderma lucidum extract and camellia oil are mixed with excipients such as microcrystalline cellulose, pregelatinized starch, magnesium stearate, and silicon dioxide, and the mixture is passed through a 40-60 mesh sieve to obtain mixture 1; S2: Lycopene powder and composite stabilizer are mixed and passed through a 40-60 mesh sieve to obtain mixture 2; S3: Mix mixture 1 and mixture 2 evenly, and put them into empty capsules to obtain the product.

[0068] The rest is the same as Example 1.

[0069] Comparative Example 3 A lycopene capsule, compared with Example 1, only the weight fractions of the raw materials are changed, specifically as follows: 35 parts of lycopene powder, 68 parts of pyrroloquinoline quinone disodium salt and 15 parts of composite stabilizer.

[0070] The rest is the same as Example 1.

[0071] Comparative Example 4 A lycopene capsule, compared with Example 1, uses sodium D-isoascorbate instead of the composite stabilizer, specifically as follows: Calculated by weight, it includes the following raw materials: 100 parts of lycopene powder, 10 parts of pyrroloquinoline quinone disodium salt and 8 parts of sodium D-isoascorbate.

[0072] The preparation method of lycopene capsules is as follows: S1: Pyrroloquinoline quinone disodium salt is mixed with excipients such as microcrystalline cellulose, pregelatinized starch, magnesium stearate, and silicon dioxide, and the mixture is passed through a 40-60 mesh sieve to obtain a mixture 1; S2: Lycopene powder and sodium D-isoascorbate were mixed and passed through a 40-60 mesh sieve to obtain mixture 2; S3: Mix mixture 1 and mixture 2 evenly, and put them into empty capsules to obtain the product.

[0073] The specification of lycopene capsules is 0.3g / capsule.

[0074] Comparative Example 5 A lycopene capsule, compared with Example 1, uses emblica extract instead of the composite stabilizer, specifically as follows: 100 parts of lycopene powder, 10 parts of pyrroloquinoline quinone disodium salt and 8 parts of emblica officinalis extract.

[0075] The preparation method of lycopene capsules is as follows: S1: Pyrroloquinoline quinone disodium salt is mixed with excipients such as microcrystalline cellulose, pregelatinized starch, magnesium stearate, and silicon dioxide, and the mixture is passed through a 40-60 mesh sieve to obtain a mixture 1; S2: Lycopene powder and emblica extract are mixed and passed through a 40-60 mesh sieve to obtain mixture 2; S3: Mix mixture 1 and mixture 2 evenly, and put them into empty capsules to obtain the product.

[0076] The rest is the same as Example 1.

[0077] Comparative Example 6 A lycopene capsule, compared with Example 1, uses Echinacea extract instead of the composite stabilizer, specifically as follows: 100 parts lycopene powder, 10 parts pyrroloquinoline quinone disodium salt and 8 parts echinacea extract.

[0078] The preparation method of lycopene capsules is as follows: S1: Pyrroloquinoline quinone disodium salt is mixed with excipients such as microcrystalline cellulose, pregelatinized starch, magnesium stearate, and silicon dioxide, and the mixture is passed through a 40-60 mesh sieve to obtain a mixture 1; S2: Lycopene powder and Echinacea extract are mixed and passed through a 40-60 mesh sieve to obtain mixture 2; S3: Mix mixture 1 and mixture 2 evenly, and put them into empty capsules to obtain the product.

[0079] The rest is the same as Example 1.

[0080] Test Example 1 Stability test: The lycopene content of the pre-packaged soft capsules in each Example and Comparative Example was determined with reference to GB / T 22249-2008 "Determination of Lycopene in Health Foods." Thirty packaged soft capsules from each Example and Comparative Example were collected and stored sealed and uncovered at 50°C and 60% relative humidity for four months. The lycopene content of ten soft capsules from each group was determined at one, three, and four months. The lycopene loss rate was calculated and the average value was taken.

[0081] Lycopene loss rate = (initial lycopene content - lycopene content after storage) / initial lycopene content.

[0082] The results are shown in Table 1 below: Table 1. Stability test results

[0083] Test Example 2 Test of the effect of lycopene capsules on immunity: 1. Materials and Methods 1.1 Sample: Lycopene capsules prepared in Example 1 and Comparative Examples 1 to 3. The test substances were prepared with sterile water and prepared immediately before use.

[0084] 1.2 Experimental Animals: 120 SPF-grade KM female mice weighing 18-22 g were bred by Beijing Huafukang Biotechnology Co., Ltd. (License No.: SCXK-(Beijing)-2024-0003). After 3 days of acclimatization and observation, the mice were randomly divided into 15 groups according to body weight, with 8 mice in each group, corresponding to different doses: Blank control group; Low-dose groups (Example 1 group, Comparative Example 1 group to Comparative Example 3 group); Medium dose (Example 1 group, Comparative Example 1 group to Comparative Example 3 group); High dose (Example 1-Example 3 groups, Comparative Example 1-Comparative Example 3 groups).

[0085] The organ / body weight ratio, delayed hypersensitivity test, and half hemolysis value (HC 50 ) and determination of the number of antibody-producing cells.

[0086] Laboratory Animal Quality Certificate: No. 110322241101639721. Experimental animals were housed in an SPF-grade animal room at the Health Food Functional Testing Center, College of Applied Arts and Sciences, Beijing Union University (conditions: temperature 20°C–26°C, relative humidity 30%–70%). Laboratory Animal Use License Number: SYXK(Beijing)2023-0013. Maintenance feed was produced by Beijing Huafukang Biotechnology Co., Ltd. [License Number: SCXK(Beijing)2024-0003].

[0087] 1.3 Dosage: The recommended human dose of the lycopene capsules prepared in the present invention is 0.6g / 60kg BW per day. The low, medium, and high dose groups were set up with doses 5, 10, and 30 times the recommended human dose, i.e., 0.05g / kg BW, 0.10g / kg BW, and 0.30g / kg BW per day, respectively.

[0088] High-dose test substance: weigh 1.50 g of sample and add sterile water to 50.0 mL; Medium-dose test substance: weigh 0.50 g of sample and add sterile water to 50.0 mL; Low-dose test substance: Weigh 0.25 g of sample and add sterile water to 50.0 mL.

[0089] The samples are capsules prepared in various examples or comparative examples, and the capsule shells are removed to serve as samples.

[0090] The test substance was orally administered once daily for 31 consecutive days, and then various indicators were measured. The oral volume of the mice was 10 mL / kg BW.

[0091] A blank control group (0 g / kg BW) was also established, in which sterile water was used instead of the test substance, and the daily gavage volume was the same as that of the test substance groups. All dose groups were given maintenance feed.

[0092] 1.4 Main instruments and reagents: BP211D electronic balance (2004013), T1000 electronic balance (2012003), BS223S electronic balance (2008007), BS2202S electronic balance (2014007), UV2600 ultraviolet-visible spectrophotometer (2015003), SPARK microplate reader (2021001), carbon dioxide incubator, centrifuge, constant temperature water bath, microscope, micrometer screw (96099).

[0093] Cell counter, 24-well and 96-well flat-bottom cell culture plates, 96-well U-shaped cell culture plates, sheep red blood cells (SRBC), normal saline, Hank's solution (pH 7.2-7.4), RPMI1640 culture medium, calf serum, concanavalin A (ConA), 1 mol / L HCl solution, acidic isopropyl alcohol (96 mL isopropyl alcohol plus 4 mL 1 mol / L HCl solution), MTT, PBS buffer (pH 7.2-7.4), complement (guinea pig serum), SA buffer, agarose, Hb dilution kit, YAC-1 cells, lithium lactate, nitrotetrazolium chloride, phenazine dimethyl sulfate, oxidized coenzyme I, 0.2 mol / L Tris-HCl buffer (pH 8.2), 1% NP40, India ink, 0.1% Na2CO3 solution, chicken red blood cells, methanol, Giemsa stain, etc.

[0094] 1.5 Experimental methods: 1.5.1 Determination of body weight and organ-to-body weight ratio Weigh the mice before and after the experiment to observe the effects of the test substances on their body weight. On the day of the experiment, the mice were weighed and sacrificed by cervical dislocation. The spleen and thymus were removed, the fascia was removed, and the blood on the surface of the organs was blotted with filter paper. The organs were weighed, and the spleen / body weight ratio and thymus / body weight ratio were calculated.

[0095] 1.5.2 ConA-induced mouse lymphocyte transformation assay (MTT method) Remove the spleen aseptically and place it in a small dish containing an appropriate amount of sterile Hank's solution. Use forceps to gently grind the spleen to prepare a single cell suspension. Filter through a 200-mesh sieve to prepare a cell suspension. Wash twice with Hank's solution, centrifuging each time for 10 minutes (1000 rpm). Then suspend the cells in 1 mL of complete culture medium and count them under a microscope to adjust the cell concentration to 3 × 10 6 The spleen cell suspension was then added to two wells of a 24-well culture plate, 1.0 mL per well. 75 μL of ConA solution (equivalent to 7.5 μg / mL) was added to one well, while the other well served as a control. The cells were incubated in a CO2 incubator at 37°C in a 5% CO2 atmosphere for 72 hours. Four hours before the end of the incubation period, 0.7 mL of supernatant was gently aspirated from each well, and 0.7 mL of RPMI-1640 culture medium without calf serum was added. MTT (5 mg / mL) was also added at 50 μL / well, and the cells were incubated for another 4 hours. After the incubation period, 1.0 mL of acidic isopropanol was added to each well and mixed by pipetting to completely dissolve the purple crystals. This solution was then transferred to a cuvette and the OD value was measured on a spectrophotometer at a wavelength of 570 nm. The proliferation capacity of lymphocytes was determined by subtracting the optical density of the well without ConA from the optical density of the well with ConA. A positive result was determined if the optical density difference of the test sample group was significantly higher than that of the control group.

[0096] 1.5.3 Delayed-type hypersensitivity (DTH) test (plantar thickening method) Defibrinated sheep blood was collected and washed three times with saline. Each mouse was then injected intraperitoneally with 0.2 mL of a 2% (v / v, prepared with saline) SRBC suspension (2000 rpm, 10 minutes). Four days after sensitization, the thickness of the left hind paw was measured. Then, 20 μL of a 20% (v / v, prepared with saline) SRBC suspension was injected subcutaneously at the measurement site. The thickness of the left hind paw was measured 24 hours after injection. Both measurements were performed three times at the same site, and the average value was taken. The difference in paw thickness before and after the attack was used to represent the degree of DTH. If the difference in the test sample group was significantly higher than that in the control group, the test result was considered positive.

[0097] 1.5.4 Determination of Antibody-Producing Cell Numbers (Jerne Modified Slide Method) Defibrinated sheep blood was collected and washed three times with saline. Each mouse was intraperitoneally injected with 0.2 mL of a 2% (v / v, prepared in saline) SRBC suspension. Five days after SRBC immunization, mice were sacrificed by cervical dislocation. The spleen was removed and gently ground to prepare a cell suspension (filtered through a 200-mesh sieve). The cells were centrifuged (1000 rpm) for 10 minutes, washed twice with Hank's solution, and finally suspended in 8.0 mL of Hank's solution. Dissolve 1% agarose by heating, then mix with an equal volume of double Hank's solution. Aliquot 0.5 mL into small test tubes. Add 50 μL of a 10% (v / v) SRBC suspension prepared with SA buffer and 25 μL of a spleen cell suspension to each tube. Mix quickly and pour onto a glass slide coated with a thin layer of agarose. Create parallel sections. After the agar solidifies, place the slide horizontally on a slide rack and incubate in a CO2 incubator at 37°C for 1 hour. Then, add complement diluted in SA buffer (1:8) into the grooves of the slide rack. After a further incubation of 1.5 hours, count the hemolytic plaques. This reflects the number of antibody-producing cells and is expressed as plaques / total splenocytes. A significantly higher number of plaques in the test sample group than in the control group indicates a positive result.

[0098] 1.5.5 Half hemolysis value (HC 50 ) Defibrinated sheep blood was collected and washed three times with saline. Each mouse was immunized with 0.2 mL of a 2% (v / v) SRBC suspension prepared in saline. Five days later, the eyeballs were enucleated and blood was collected in a centrifuge tube. The tube was allowed to stand for approximately 1 hour, and the coagulated blood was separated from the tube wall to allow the serum to fully precipitate. The tube was then centrifuged at 3000 rpm for 10 minutes to collect the serum. The serum was diluted 300-fold with SA buffer, and 1.0 mL was transferred to a test tube. 0.5 mL of a 10% (v / v) SRBC suspension prepared in SA buffer and 1.0 mL of complement (diluted 1:8 in SA buffer) were then added. A control tube without serum (using SA buffer) was also prepared. The tube was incubated in a 37°C water bath for 15 minutes, and then the reaction was terminated by ice bath. The tube was centrifuged at 2000 rpm for 10 minutes, and 1.0 mL of the supernatant was collected and added to 3.0 mL of Hb diluent. At the same time, take 0.25 mL of 10% (v / v, prepared with SA buffer) SRBC suspension, add Hb diluent to 4.0 mL in another test tube, mix thoroughly, let stand for 10 minutes, and measure the optical density of each tube at 540 nm with the control tube as blank. The amount of hemolysin is calculated as the half hemolysis value (HC 50 ) is expressed as follows: The half-hemolysis value of the sample = the optical density value of the sample / the optical density value of the SRBC when half of the hemolysis is achieved × the dilution factor.

[0099] The HC50 of the test sample group was significantly higher than that of the control group. 50, the test result can be determined to be positive.

[0100] 1.6 Data Processing SPSS software was used for data processing. ANOVA was used, but the homogeneity test of variance was performed first according to the procedure of ANOVA. If the variance was homogeneous, the F value was calculated. <F 0.06 Conclusion: There is no significant difference in the mean values ​​of each group; F value ≥ F 0.06 , P ≤ 0.05, and the pairwise comparison method between the means of multiple experimental groups and a control group was used for statistical analysis; for non-normal or heterogeneous variance data, appropriate variable transformation was performed, and after the normality or homogeneous variance requirements were met, the transformed data were used for statistical analysis; if the normality or homogeneous variance requirements were still not met after the variable transformation, the rank sum test was used for statistical analysis.

[0101] 2. Results 2.1 Effects of the test substances on mouse body weight Table 1. Initial body weight of mice ( )

[0102] As can be seen from the data in Table 1, there was no significant difference in the initial body weight of mice between each dosage group and the 0 g / kg BW group, and there was no significant difference between the examples and the comparative examples.

[0103] Table 2. Effects of different doses of the test substances on body weight ( )

[0104] As can be seen from the data in Table 2, there were no significant differences between each dosage group and the 0 g / kg BW group, and there were no significant differences between the examples and the comparative examples.

[0105] 2.2 Effects of the test substances on the organ / body weight ratio of mice Table 3. Effects of different doses of the test substance on the spleen / body weight ratio ( )

[0106] * indicates P < 0.05 compared with the blank group, and ** indicates P < 0.01 compared with the blank group.

[0107] From the data in Table 3, it can be seen that the spleen / body weight ratios of Comparative Examples 1 to 3 are significantly different compared with those of the Examples.

[0108] Table 4. Effects of different doses of the test substance on the thymus / body weight ratio ( )

[0109] * indicates P < 0.05 compared with the blank group, and ** indicates P < 0.01 compared with the blank group.

[0110] From the data in Table 4, it can be seen that the thymus / body weight ratios of Comparative Examples 1 to 3 are significantly different compared with those of the Examples.

[0111] 2.3 Effects of the test substances on the cellular immune function of mice Table 5. Effects of different doses of the test substance on delayed allergic reactions ( )

[0112] Note: * indicates P < 0.05 compared with 0.00 g / kg BW; ** indicates P < 0.01 compared with 0.00 g / kg BW.

[0113] As can be seen from the data in Table 5, after oral administration of different doses of the test substance to mice for 31 days, the paw swelling of the mice in each dose group of the Example was increased compared with the 0 g / kg BW group, with significant differences.

[0114] Table 6. Effects of different doses of test substances on lymphocyte transformation assay (x±SD)

[0115] Note: * indicates P < 0.05 compared with 0.00 g / kg BW; ** indicates P < 0.01 compared with 0.00 g / kg BW.

[0116] As shown in Table 6, after oral administration of different doses of the test substance to mice for 31 days, the lymphocyte proliferation ability of mice in each dose group of the embodiment was significantly improved compared with that in the 0 g / kg BW group.

[0117] 2.4 Effects of the test substances on the humoral immunity of mice Table 7. Effects of different doses of the test substance on the number of antibody-producing cells (x±SD)

[0118] Note: * indicates P < 0.05 compared with 0.00 g / kg BW; ** indicates P < 0.01 compared with 0.00 g / kg BW.

[0119] As shown in Table 7, after oral administration of different doses of the test substance to mice for 31 days, the number of hemolytic plaques in the mice in each dose group of the Example increased compared with the 0 g / kg BW group, with significant differences.

[0120] Table 8 Effects of different doses of the test substance on the median hemolysis value (x±SD)

[0121] Note: * indicates P < 0.05 compared with 0.00 g / kg BW; ** indicates P < 0.01 compared with 0.00 g / kg BW.

[0122] As shown in Table 8, after oral administration of different doses of the test substance to mice for 31 days, the median hemolysis value of mice in each group of the Example was increased compared with the 0 g / kg BW group, with significant differences.

[0123] Finally, it should be noted that the above content is only used to illustrate the technical solution of the present invention, rather than to limit the scope of protection of the present invention. Simple modifications or equivalent substitutions of the technical solution of the present invention by ordinary technicians in this field do not deviate from the essence and scope of the technical solution of the present invention.

Claims

1. A lycopene capsule, characterized in that: The invention comprises the following raw materials: lycopene powder, pyrroloquinoline quinone disodium salt and a composite stabilizer, wherein the composite stabilizer is a mixture of emblica fruit extract, echinacea purpurea extract and sodium D-isoascorbate.

2. The lycopene capsule according to claim 1, characterized in that The mass ratio of the emblica fruit extract, the echinacea purpurea extract and sodium D-isoascorbate is 10-20:1-10:1-3.

3. The lycopene capsule according to claim 2, characterized in that The mass ratio of the emblica fruit extract, the echinacea purpurea extract and sodium D-isoascorbate is 12-18:1-8:1-2.

4. The lycopene capsule according to claim 1, characterized in that The preparation method of the emblica extract comprises the following steps: mixing the emblica with water, heating and refluxing for 1-3 times to obtain a water extract and residues, gradient alcohol extraction of the residues to obtain an alcohol extract, mixing the water extract and the alcohol extract, and drying to obtain the emblica extract.

5. The lycopene capsule according to claim 4, characterized in that The gradient alcohol extraction step is as follows: the residue is mixed with a 35wt%-45wt% ethanol solution for a first alcohol extraction, then the concentration of the ethanol solution is increased to 50wt%-65wt% for a second alcohol extraction, then the concentration of the ethanol solution is increased to 75wt%-85wt% for a third alcohol extraction, and the mixture is combined to obtain an alcohol extract.

6. The lycopene capsule according to claim 1, characterized in that The preparation method of the Echinacea extract comprises the following steps: mixing Echinacea with alcohol, performing alcohol extraction at 40-50° C. for 1-3 times, combining the alcohol extracts, and drying.

7. The lycopene capsule according to claim 6, characterized in that The alcohol is a 75wt%-90wt% ethanol solution.

8. The lycopene capsule according to claim 6, characterized in that The material-liquid ratio of Echinacea to alcohol is 1:8-12 g:L.

9. The lycopene capsule according to claim 1, characterized in that The lycopene capsules include the following raw materials in parts by mass: 40-160 parts of lycopene powder, 1-10 parts of pyrroloquinoline quinone disodium salt and 1-12 parts of a composite stabilizer.

10. The lycopene capsule according to claim 9, characterized in that The lycopene capsules include the following raw materials in parts by mass: 50-150 parts of lycopene powder, 2-10 parts of pyrroloquinoline quinone disodium salt and 1-10 parts of a composite stabilizer.

11. The lycopene capsule according to claim 1, characterized in that The lycopene capsules may further include auxiliary materials.

12. The lycopene capsule according to claim 11, characterized in that The auxiliary materials include, but are not limited to, excipients, buffers, emulsifiers, diluents, binders, preservatives, lubricants, pH adjusters, cryoprotectants, flavorings, fillers and antioxidants.

13. The method for preparing lycopene capsules according to any one of claims 1 to 12, characterized in that: The following steps are involved: S1: Pyrroloquinoline quinone disodium salt is mixed with excipients to obtain mixture 1; S2: Lycopene powder is mixed with a composite stabilizer to obtain mixture 2; S3: Mix mixture 1 and mixture 2, and put them into empty capsules to obtain the product.

14. Use of the lycopene capsule according to any one of claims 1 to 12 in the preparation of health food for improving immunity.

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