阅读说明：本技术 一种速溶型纳米级生物素微胶囊及其制备方法和应用 (Instant nano-scale biotin microcapsule and preparation method and application thereof ) 是由 魏初权 刘锦洪 王文积 林木荣 王雪瑞 于 2021-09-27 设计创作，主要内容包括：本发明属于生物素技术领域,具体涉及一种速溶型纳米级生物素微胶囊及其制备方法和应用。所述速溶型纳米级生物素微胶囊包括囊芯和囊材,所述囊芯包覆于囊材中,以速溶型纳米级生物素微胶囊的总重量为基准,所述囊芯中含有5～15％生物素结晶、20～30％载体油和0.5～1.5％油相乳化剂,所述囊材中含有20～40％水相乳化壁材、10～50％小分子填充物、2～5％助溶剂和0.5～1.5％稳定剂,所述生物素结晶的晶体粒径≤100nm,所述小分子填充物的分子量≤1000。本发明提供的纳米级生物素微胶囊为速溶型,耐酸性强,溶解冲调性好,有利于肠道吸收,生物利用度高。(The invention belongs to the technical field of biotin, and particularly relates to an instant nano-scale biotin microcapsule as well as a preparation method and application thereof. The instant nano-scale biotin microcapsule comprises a capsule core and a capsule wall material, wherein the capsule core is coated in the capsule wall material, the capsule core contains 5-15% of biotin crystals, 20-30% of carrier oil and 0.5-1.5% of oil phase emulsifier by taking the total weight of the instant nano-scale biotin microcapsule as a reference, the capsule wall material contains 20-40% of water phase emulsifying wall materials, 10-50% of micromolecular fillers, 2-5% of cosolvent and 0.5-1.5% of stabilizing agents, the crystal grain size of the biotin crystals is less than or equal to 100nm, and the molecular weight of the micromolecular fillers is less than or equal to 1000. The nano-scale biotin microcapsule provided by the invention is instant, has strong acid resistance and good dissolution and reconstitution properties, is beneficial to intestinal absorption, and has high bioavailability.)

1. The instant nano-scale biotin microcapsule is characterized by comprising a capsule core and a capsule wall material, wherein the capsule core is coated in the capsule wall material, the capsule core contains 5-15% of biotin crystals, 20-30% of carrier oil and 0.5-1.5% of oil phase emulsifier by taking the total weight of the instant nano-scale biotin microcapsule as a reference, the capsule wall material contains 20-40% of water phase emulsifying wall materials, 10-50% of small molecule fillers, 2-5% of cosolvent and 0.5-1.5% of stabilizing agents, the crystal grain size of the biotin crystals is less than or equal to 100nm, and the molecular weight of the small molecule fillers is less than or equal to 1000.

2. The fast-dissolving nanoscale biotin microcapsule according to claim 1,

the carrier oil is vegetable oil and/or fatty acid ester; the vegetable oil is preferably at least one selected from corn oil, soybean oil, peanut oil, olive oil, sesame oil, sunflower seed oil, safflower oil, linseed oil, castor oil, cottonseed oil, hemp seed oil, linseed oil, camellia seed oil, peony seed oil, palm oil, walnut oil, coconut oil, rice oil and rapeseed oil; the fatty acid ester is preferably at least one selected from the group consisting of caproic acid triglyceride, caprylic acid triglyceride, capric acid triglyceride, caprylic acid triglyceride, enanthic acid triglyceride and lauric acid triglyceride;

preferably, the oil phase emulsifier is at least one of sucrose ester with HLB value of 5-7, vitamin E polyethylene glycol succinate, glyceride and lecithin;

preferably, the water phase emulsifying wall material is selected from at least one of modified starch, Arabic gum, whey protein, sodium caseinate and soy protein isolate; the modified starch is preferably at least one selected from acid modified starch, starch octenylsuccinate, starch sodium octenylsuccinate, oxidized starch, acetate starch, hydroxypropyl starch and pregelatinized starch;

preferably, the small molecular filler is selected from at least one of monosaccharide, disaccharide and oligosaccharide with the molecular weight of 150-500, preferably at least one of glucose, maltose, sucrose, maltodextrin, solid corn syrup and resistant dextrin;

preferably, the cosolvent is selected from at least one of glycerol, sorbitol, mannitol, and hexaglycerol laurate;

preferably, the stabilizer is selected from at least one of sodium hexametaphosphate, potassium tripolyphosphate, trisodium citrate, ethylenediaminetetraacetic acid and sodium salts thereof, ascorbic acid and sodium salts thereof.

3. The instant nano-scale biotin microcapsule according to claim 1, further comprising a pigment and/or a flavor, wherein the oil-soluble component of the pigment and the flavor is distributed in the core and the water-soluble component of the pigment and the flavor is distributed in the capsule wall material.

4. The instant nano-sized biotin microcapsule according to claim 1, having a particle size D₉₀Is 300 to 500 μm.

5. The instant nano-sized biotin microcapsule according to claim 1, wherein 1g of the instant nano-sized biotin microcapsule is dissolved in 100mL of water at room temperature to give an emulsion having a particle size distribution D of oil droplets₁₀≤0.15μm，D₅₀≤0.5μm，D₉₀≤1μm。

6. The method for preparing the instant nano-scale biotin microcapsules according to any one of claims 1 to 5, comprising the steps of:

s1, stirring and dispersing the biotin crystals, an oil phase emulsifier and optional oil-soluble components in pigments and/or flavors and fragrances in carrier oil, then grinding until the particle size of the crystals of the biotin crystals is less than or equal to 100nm, and then cooling the system to 0-20 ℃ to obtain an oil suspension I;

s2, stirring and dissolving water-soluble components in the water-phase emulsification wall material, the small-molecule filler, the cosolvent, the stabilizer and the optional pigment and/or essence perfume in water at 50-60 ℃, and then cooling the system to 0-20 ℃ to obtain a water phase II;

s3, mixing and homogenizing the oil suspension I and the water phase II at 0-20 ℃, and adjusting the viscosity to 100-350 mPa & S to obtain an emulsion;

and S4, spray drying the emulsion to obtain the instant nano-scale biotin microcapsule.

7. The method for preparing the instant nano-scale biotin microcapsules according to claim 6, wherein in step S3, the oil suspension I and the water phase II are mixed and homogenized, and then the temperature of the system is controlled to be 0-20 ℃ and the shearing speed is controlled to be 8000-12000 r/min for shearing emulsification for 10-30 min so as to make the diameter D of oil drops be larger than the diameter D of the oil drops₉₀Less than or equal to 2 mu m, and then homogenizing for 2-3 times under the high pressure of 30-60 MPa to ensure that the grain diameter D of the oil drops₉₀≤1μm。

8. The method for preparing the instant nano-scale biotin microcapsule according to claim 6, wherein the spray drying conditions in step S4 include an inlet air temperature of 100-180 ℃, an outlet air temperature of 50-90 ℃, and an inlet air volume of 20-30 m³The pressure of the spray tower is 0.1-0.5 KPa.

9. The method for preparing the instant nano-scaled biotin microcapsule according to claim 6, further comprising adding an anti-caking agent to the instant nano-scaled biotin microcapsule after the spray-drying; the anticaking agent is selected from at least one of silicon dioxide, tricalcium phosphate, calcium silicate and microcrystalline cellulose; the dosage of the anticaking agent is not more than 1 wt% of the mass of the instant nano-scale biotin microcapsule.

10. Use of the instant nano-scale biotin microcapsules of any one of claims 1 to 5 as a food additive or a pharmaceutical product.

Technical Field

The invention belongs to the technical field of biotin, and particularly relates to an instant nano-scale biotin microcapsule as well as a preparation method and application thereof.

Background

Biotin (Biotin), also known as vitamin H and coenzyme r (coenzyme r), is a water-soluble vitamin, also belongs to the vitamin B group, specifically vitamin B7, is an essential substance for synthesizing vitamin C, is an essential substance for normal metabolism of fats and proteins, and is a nutrient necessary for maintaining the natural growth, development and normal function of the human body. Biotin was found in the liver as a factor for preventing and treating depilation and skin damage of rats induced by feeding raw egg protein in the study of growth and respiration promoting factors of yeast growth factors and rhizobia in the 30 s of the 20 th century. Biotin is easily bound to one of the chicken proteins, and a large amount of dietary raw protein can prevent absorption of biotin, resulting in biotin deficiency, such as alopecia, weight loss, dermatitis, etc. Biotin plays an important role in biochemical reaction pathways such as fat synthesis and carbohydrate regeneration. Biotin is a saving star of baldness, has a good effect on preventing hair loss and light on the top of the head, and can also prevent the common juvenile canities of modern people. It also plays an important role in maintaining skin health, and is beneficial to depression and insomnia.

Biotin is colorless needle crystal, is very slightly soluble in water (22mg/100mL water, 25 deg.C) and ethanol (80mg/100mL, 25 deg.C), is insoluble in other common organic solvents, is stable to heat, and can be destroyed by strong acid, strong base and oxidant, and can be gradually destroyed by ultraviolet ray. Biotin is mainly stored in the liver and is present in the blood at a low level. The biotin in the food exists mainly in a free form or a form combined with protein, and the biotin combined with the protein forms biocytin under the action of intestinal protease, and then releases the free biotin through the action of intestinal biotinidase. The main part of biotin absorption is the proximal end of the small intestine, and the biotin is actively absorbed by carrier transport at low concentration; at high concentration, biotin is absorbed in a simple diffusion mode, absorbed biotin is circulated through portal vessels and transported to the liver and kidney for storage, and other cells also contain biotin but in a small amount. Human intestinal bacteria can synthesize biotin from diopimelic acid instead of pelargonic acid, but the direct source of biotin is not enough. Most of biotin products in the market are pure products or pure products and other auxiliary materials are simply mixed for use, the bioavailability is low, and the improvement of the bioavailability becomes the direction of the current industry exploration.

Although patent documents and non-patent documents are commonly reported about biotin application and synthesis methods, no reports about biotin microencapsulation technology are found, because biotin is non-fat-soluble and is very slightly soluble in water (22mg/100mL water, 25 ℃) although being a water-soluble vitamin, and has low solubility in hot water and high difficulty in microencapsulation, and thus no reports about biotin microencapsulation technology are found at present.

Disclosure of Invention

The invention aims to overcome the defect of low bioavailability of the existing biotin product, and provides a quick-dissolving nano-scale biotin microcapsule capable of improving bioavailability and a preparation method and application thereof.

After intensive research, the inventor of the present invention finds that the main reason for the low bioavailability of the biotin products in the market, which are used in the form of pure products or pure products simply mixed with other auxiliary materials, is that: on one hand, the biotin crystal particles in the biotin products are large, poor in dissolving and dissolving performance and poor in stability, and are difficult to absorb and utilize by organisms; on the other hand, the biotin products are not resistant to strong acid, the pH value of gastric juice is 0.9-1.8 (strong acid), the pH value of small intestine liquid is 8.0-9.0 (alkaline), the main part of biotin absorption is small intestine, and when the biotin is taken in a form of pure product or simple mixture of the pure product and other auxiliary materials, the biotin is destroyed in the strong acid gastric juice and cannot effectively reach the intestinal tract, so that the bioavailability is low, and the health care or treatment effect is poor.

After intensive research, the inventor of the invention finds that after biotin is prepared into microcapsule powder, the solubility of the biotin can be improved, the biotin is protected from the destructive effect of strong acid in gastrointestinal tract, and the absorption of the biotin is enhanced. However, as mentioned above, the key reason why no reports on the biotin microencapsulation technology are available at present is that the unique property of non-fat solubility and very slight water solubility of biotin results in great difficulty in microencapsulation. In the process of biotin microencapsulation, non-fat-soluble biotin which is extremely slightly dissolved in water is dispersed in carrier oil in the presence of an oil phase emulsifier and is firstly prepared into oil suspension of nano microcrystals through nano grinding treatment, and then the oil suspension is prepared into microcapsule powder through microencapsulation embedding by adopting a low-temperature emulsification method, so that the effective coating of the biotin microcrystals can be realized, the solubility of the biotin can be effectively improved, the biotin is protected from the destructive effect of strong acid in gastrointestinal tracts, and the bioavailability of the biotin can be enhanced. Based on this, the present invention has been completed.

The invention provides an instant nano-scale biotin microcapsule, which comprises a capsule core and a capsule wall material, wherein the capsule core is coated in the capsule wall material, the capsule core contains 5-15% of biotin crystals, 20-30% of carrier oil and 0.5-1.5% of oil-phase emulsifier by taking the total weight of the instant nano-scale biotin microcapsule as a reference, the capsule wall material contains 20-40% of water-phase emulsifying wall material, 10-50% of small-molecule filler, 2-5% of cosolvent and 0.5-1.5% of stabilizer, the crystal grain size of the biotin crystals is less than or equal to 100nm, and the molecular weight of the small-molecule filler is less than or equal to 1000.

Further, the carrier oil is a vegetable oil and/or a fatty acid ester.

Further, the vegetable oil is selected from at least one of corn oil, soybean oil, peanut oil, olive oil, sesame oil, sunflower oil, safflower oil, linseed oil, castor oil, cottonseed oil, hemp oil, linseed oil, camellia seed oil, peony seed oil, palm oil, walnut oil, coconut oil, rice oil and rapeseed oil.

Further, the fatty acid ester is selected from at least one of caproic acid triglyceride, caprylic acid triglyceride, capric acid triglyceride, caprylic acid triglyceride, heptanoic acid triglyceride and lauric acid triglyceride.

Further, the oil phase emulsifier is at least one of sucrose ester with HLB value of 5-7, vitamin E polyethylene glycol succinate, glyceride and lecithin.

Further, the water phase emulsifying wall material is at least one selected from modified starch, Arabic gum, whey protein, sodium caseinate and isolated soy protein, and is preferably modified starch.

Further, the modified starch is selected from at least one of acid-modified starch, starch octenyl succinate, starch sodium octenyl succinate, oxidized starch, starch acetate, hydroxypropyl starch, and pregelatinized starch.

Further, the small molecular filler is selected from at least one of monosaccharide, disaccharide and oligosaccharide with the molecular weight of 150-500, and is preferably selected from at least one of glucose, maltose, sucrose, maltodextrin, solid corn syrup and resistant dextrin.

Further, the cosolvent is at least one selected from glycerol, sorbitol, mannitol and hexaglycerol laurate.

Further, the stabilizer is selected from at least one of sodium hexametaphosphate, potassium tripolyphosphate, trisodium citrate, ethylene diamine tetraacetic acid and sodium salts thereof, ascorbic acid and sodium salts thereof.

Furthermore, the instant nano-scale biotin microcapsule also contains pigment and/or essence, wherein oil-soluble components in the pigment and the essence are distributed in the capsule core, and water-soluble components in the capsule wall are distributed in the capsule wall material.

Further, the particle size D of the instant nano-scale biotin microcapsule₉₀Is 300 to 500 μm.

Further, after 1g of the instant nano-scale biotin microcapsule is dissolved in 100mL of normal-temperature water, the particle size distribution D of oil drops in the emulsion is obtained₁₀≤0.15μm，D₅₀≤0.5μm，D₉₀≤1μm。

The invention also provides a preparation method of the instant nano-scale biotin microcapsule, which comprises the following steps:

s1, stirring and dispersing the biotin crystals, an oil phase emulsifier and optional oil-soluble components in pigments and/or flavors and fragrances in carrier oil, then grinding until the particle size of the crystals of the biotin crystals is less than or equal to 100nm, and then cooling the system to 0-20 ℃ to obtain an oil suspension I;

s2, stirring and dissolving water-soluble components in the water-phase emulsification wall material, the small-molecule filler, the cosolvent, the stabilizer and the optional pigment and/or essence perfume in water at 50-60 ℃, and then cooling the system to 0-20 ℃ to obtain a water phase II;

s3, mixing and homogenizing the oil suspension I and the water phase II at 0-20 ℃, and adjusting the viscosity to 100-350 mPa & S to obtain an emulsion;

and S4, spray drying the emulsion to obtain the instant nano-scale biotin microcapsule.

Further, in step S3, the mixing and homogenizing manner is to mix the oil suspension I and the water phase II, and then the system temperature is controlled to be 0-20 ℃ and the shearing rotation speed is controlled to be 8000-12000 r/min for shearing and emulsifying for 10-30 min so as to ensure thatTo obtain the oil drop particle diameter D₉₀Less than or equal to 2 mu m, and then homogenizing for 2-3 times under the high pressure of 30-60 MPa to ensure that the grain diameter D of the oil drops₉₀≤1μm。

Further, in the step S4, the spray drying conditions include an inlet air temperature of 100-180 ℃, an outlet air temperature of 50-90 ℃, and an inlet air volume of 20-30 m³The pressure of the spray tower is 0.1-0.5 KPa.

Furthermore, the preparation method of the instant nano-scale biotin microcapsule provided by the invention also comprises the step of adding an anticaking agent into the instant nano-scale biotin microcapsule after spray drying.

Further, the anti-caking agent is selected from at least one of silicon dioxide, tricalcium phosphate, calcium silicate and microcrystalline cellulose.

Further, the dosage of the anticaking agent is not more than 1 wt% of the mass of the instant nano-scale biotin microcapsule.

The invention also provides application of the instant nano-scale biotin microcapsule as a food additive or a medicine.

The key point of the invention is that the biotin is made into the microcapsule by adopting a specific microencapsulation technology, specifically, the biotin crystal particles are dispersed in carrier oil and are firstly made into oil suspension of nano microcrystals through nano grinding treatment, and then the oil suspension is microencapsulated and embedded by adopting a low-temperature emulsification method to prepare microcapsule powder, so that the obtained microcapsule has better solubility and acid resistance, can protect the biotin from the destructive effect of strong acid in stomach, can effectively reach the small intestine to be effectively absorbed, enhances the bioavailability of the biotin and can directly carry related components to enter blood. The instant nano-scale biotin microcapsule provided by the invention has a wide pH application range, can be used at a pH value of 1-14, and expands the application field of biotin products. In addition, the invention adopts a low-temperature emulsification method to prepare the instant nano-scale biotin microcapsule, can effectively prevent biotin from being dissolved by water phase in the microencapsulation process, effectively microencapsulates biotin microcrystals, and adds a cosolvent and a stabilizer in the microencapsulation process, thereby remarkably improving the solubility, the reconstitution property and the strong acid resistance of the biotin microcapsule.

The nano-scale biotin microcapsule obtained by the method has uniform content, can be mixed with water in any proportion, is instant, has acid resistance and good dissolution and reconstitution properties, can form uniform and stable solution, has high stability, can obviously inhibit crystallization of biotin, is beneficial to intestinal absorption, has high bioavailability, can improve intestinal transmission of nutrient components and provide a protection effect, can be added into systems such as foods, medicines and the like only according to the required amount in the application process, and is extremely convenient to use.

Detailed Description

In the invention, the instant nano-scale biotin microcapsule comprises a capsule core and a capsule wall material, wherein the capsule core is coated in the capsule wall material. Wherein the capsule core contains biotin crystals, carrier oil and an oil phase emulsifier. The capsule wall material contains an aqueous phase emulsification wall material, a small molecular filler, a cosolvent and a stabilizer. Particle size D of the instant nano-scale biotin microcapsule₉₀Preferably 300 to 500 μm. Furthermore, it is preferable that 1g of the instant nano-sized biotin microcapsule is dissolved in 100mL of water at normal temperature to obtain an emulsion having a particle size distribution D of oil droplets₁₀≤0.15μm，D₅₀≤0.5μm，D₉₀≤1μm。

The crystal size of biotin in the capsule core is nanometer, and is less than or equal to 100nm, such as 5nm, 10nm, 15nm, 20nm, 25nm, 30nm, 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, 65nm, 70nm, 75nm, 80nm, 85nm, 90nm, 95nm, and 100 nm. The content of the biotin crystal is 5 to 15%, and may be, for example, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%.

The carrier oil in the capsule core plays a role in suspending and dispersing the biotin crystals so as to facilitate grinding the biotin crystals to a nanometer level. The carrier oil may be a vegetable oil and/or a fatty acid ester. The vegetable oil can be natural vegetable oil, or vegetable oil after structural modification and hydrolysis, or a mixture of the two, and specifically can be at least one selected from corn oil, soybean oil, peanut oil, olive oil, sesame oil, sunflower seed oil, safflower oil, linseed oil, castor oil, cottonseed oil, hemp oil, linseed oil, camellia oil, peony seed oil, palm oil, walnut oil, coconut oil, rice oil, rapeseed oil and the like. The fatty acid ester may be at least one of caproic acid triglyceride, caprylic acid triglyceride, capric acid triglyceride, caprylic-capric acid triglyceride (MCT), heptanoic acid triglyceride and lauric acid triglyceride, preferably caprylic-capric acid triglyceride (MCT). The carrier oil has excellent mutual solubility, dilution and emulsification solubilization, can realize effective dispersion of biotin crystals, has extremely high oxidation resistance, and can avoid the change of color and smell of vegetable oil caused by rancidity. In addition, the carrier oil can be rapidly digested and absorbed without the hydrolysis of pancreatic lipase and without the participation of lymphatic system in absorption, and can promote the absorption of biotin. The content of the carrier oil is 20 to 30%, and for example, may be 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%.

The use of an oil phase emulsifier in the core ensures that a stable emulsion is formed during microencapsulation. The oil phase emulsifier can be at least one of sucrose ester with HLB value of 5-7, vitamin E polyethylene glycol succinate, glyceride and lecithin. Specific examples of the sucrose esters include, but are not limited to: sucrose acetate isobutyrate, sucrose fatty acid ester, sucrose laurate, and the like. The average molecular weight of the polyethylene glycol in the vitamin E polyethylene glycol succinate may be 200 to 8000, specifically, PEG200, PEG400, PEG800, PEG1000, PEG2000, PEG4000, PEG8000, and the like, and is not particularly limited. The glyceride may be at least one of stearic acid monoglyceride, lauric acid monoglyceride, citric acid monoglyceride, succinic acid monoglyceride, oleic acid monoglyceride, palmitic acid monoglyceride, and the like. The content of the oil phase emulsifier is 0.5 to 1.5%, and may be, for example, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%.

Specific examples of the aqueous phase emulsified wall material in the capsule material include, but are not limited to: at least one of modified starch, gum arabic, whey protein, sodium caseinate and soy protein isolate, preferably modified starch. The modified starch is preferably at least one selected from acid-modified starch, starch octenylsuccinate, starch sodium octenylsuccinate, oxidized starch, starch acetate, hydroxypropyl starch and pregelatinized starch, and the modified starch can be obtained commercially or prepared by the existing method, and the specific preparation process is well known to those skilled in the art and is not described herein. The content of the aqueous emulsification wall material is 20 to 40%, and for example, may be 20%, 21%, 22%, 23%, 24%, 25%, 26%, 27%, 28%, 29%, 30%, 31%, 32%, 33%, 34%, 35%, 36%, 37%, 38%, 39%, 40%.

The micro-molecular filler in the capsule wall material plays a role in filling micro-pores on the surface of the microcapsule, and the surface of the microcapsule can be more compact due to the good film-forming property, so that a capsule core substance (biotin) is effectively protected, and the risk that the capsule core is damaged by external conditions is reduced. The molecular weight of the small molecule filler is less than or equal to 1000, for example, the molecular weight can be 100, 150, 200, 250, 300, 350, 400, 450, 500, 550, 600, 650, 700, 750, 800, 850, 900, 950 and 1000, and is preferably 150 to 500. The small molecule filler is preferably a low molecule saccharide substance, and specifically may be at least one of monosaccharide, disaccharide and oligosaccharide, and specific examples thereof include but are not limited to: at least one of glucose, maltose, sucrose, maltodextrin, solid corn syrup, and resistant dextrin. The content of the small molecule filler is 10-50%, for example, 10%, 12%, 15%, 18%, 20%, 22%, 25%, 28%, 30%, 32%, 35%, 37%, 40%, 42%, 45%, 48%, 50%.

The cosolvent in the capsule wall material has the functions of assisting the dissolution and dispersion of the biotin microcapsules in water, promoting the dissolution performance of the biotin microcapsules in water, and inhibiting the crystallization phenomenon of biotin after the biotin microcapsules are dissolved in water, so that the absorption of the biotin by intestinal tracts is facilitated, and the bioavailability of the biotin is improved. Specific examples of such co-solvents include, but are not limited to: at least one of glycerin, sorbitol, mannitol, and hexaglycerol laurate. The content of the cosolvent is 2-5%, for example, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%.

The stabilizing agent in the capsule wall material has the functions of improving the stability of the biotin microcapsules and inhibiting the crystallization phenomenon of the biotin after the biotin microcapsules are dissolved in water, so that the absorption of the biotin by intestinal tracts is facilitated, and the bioavailability of the biotin is improved. Specific examples of the stabilizer include, but are not limited to: at least one of sodium hexametaphosphate, potassium tripolyphosphate, trisodium citrate, ethylene diamine tetraacetic acid and sodium salts thereof, ascorbic acid and sodium salts thereof. The content of the stabilizer is 0.5 to 1.5%, and may be, for example, 0.5%, 0.6%, 0.7%, 0.8%, 0.9%, 1.0%, 1.1%, 1.2%, 1.3%, 1.4%, 1.5%.

The instant nano-scale biotin microcapsule also contains pigment and/or essence and flavor, and is used for improving the color, the fragrance and the taste of the microcapsule. The types of the pigment and the essence and flavor are allowed to be used in the food field or the pharmaceutical field, the specific types can be selected conventionally in the field, and the addition amount is also the conventional addition amount allowed to be used in the food field or the pharmaceutical field, and is known to those skilled in the art, and the details are not described herein. The distribution of the pigment and the essence perfume in the microcapsule depends on the hydrophilic and lipophilic characteristics of the microcapsule, and the oil-soluble components in the pigment and the essence perfume are distributed in the capsule core and the water-soluble components in the capsule wall material.

The preparation method of the instant nano-scale biotin microcapsule provided by the invention comprises the following steps:

s1, stirring and dispersing the biotin crystals, an oil phase emulsifier and optional oil-soluble components in pigments and/or flavors and fragrances in carrier oil, then grinding until the particle size of the crystals of the biotin crystals is less than or equal to 100nm, and then cooling the system to 0-20 ℃ to obtain an oil suspension I;

s2, stirring and dissolving water-soluble components in the water-phase emulsification wall material, the small-molecule filler, the cosolvent, the stabilizer and the optional pigment and/or essence perfume in water at 50-60 ℃, and then cooling the system to 0-20 ℃ to obtain a water phase II;

s3, mixing and homogenizing the oil suspension I and the water phase II at 0-20 ℃, and adjusting the viscosity to 100-350 mPa & S to obtain an emulsion;

and S4, spray drying the emulsion to obtain the instant nano-scale biotin microcapsule.

In step S1, the biotin crystal material is commercially available, and the crystal particle size is generally not less than 50 μm. The stirring dispersion is only required to be capable of approximately dispersing the biotin crystals, the oil-phase emulsifier and optionally the oil-soluble component in the pigment and/or the essence in the carrier oil. The grinding may be performed in various existing nano-sand mills under such conditions that the crystal particle size of the biotin crystals can be reduced to 100nm or less.

In step S3, the mixing and homogenizing manner is not particularly limited as long as the oil suspension I and the water phase II can be sufficiently emulsified to form an emulsion, and preferably, the mixing and homogenizing manner is to mix the oil suspension I and the water phase II, and then, the system temperature is controlled to be 0-20 ℃, and the shearing rotation speed is controlled to be 8000-12000 r/min, and the shearing emulsification is performed for 10-30 min to make the oil drop particle diameter D₉₀Less than or equal to 2 mu m, and then homogenizing for 2-3 times under the high pressure of 30-60 MPa to ensure that the grain diameter D of the oil drops₉₀Less than or equal to 1 mu m. The mixing and homogenizing at 0-20 deg.C is selected mainly in consideration of the fact that biotin is very slightly soluble in water (22mg/100mL water, 25 deg.C), and mixing and homogenizing at a low temperature of 0-20 deg.C can reduce the probability of dissolution of biotin crystals by the water phase as much as possible, so that the biological crystals are coated in the capsule material as much as possible. In addition, the viscosity needs to be adjusted to 100-350 mPas after the mixing and homogenizing, so that the obtained emulsion can be stably carried out in the subsequent spray drying process, and a good coating effect is realized. In the present invention, the viscosity is measured using an Ubbelohde viscometer.

In the step S4, the preferable conditions of the spray drying include that the air inlet temperature is 100-180 ℃, the air outlet temperature is 50-90 ℃, and the air inlet volume is 20-30 m³The pressure of the spray tower is 0.1-0.5 KPa. In the present invention, the pressure isBoth refer to gauge pressure.

In another preferred embodiment, the method for preparing the instant nano-scale biotin microcapsules further comprises adding an anti-caking agent to the instant nano-scale biotin microcapsules after spray-drying, wherein agglomeration between the microcapsules can be effectively prevented. The anticaking agent may be at least one selected from the group consisting of silicon dioxide, tricalcium phosphate, calcium silicate, and microcrystalline cellulose. In addition, the amount of the anti-caking agent is preferably not more than 1 wt% of the mass of the instant nano-scale biotin microcapsule.

In the preparation process of the instant nano-scale biotin microcapsule, the types and the amounts of the raw materials are described above, and are not described herein again.

The invention also provides application of the instant nano-scale biotin microcapsule as a food additive or a medicine.

In the following examples and comparative examples, the components are used in parts by weight unless otherwise specified.

In the following examples and comparative examples, the content of biotin in the microcapsules was measured by HPLC, and the column was VP-ODS (4.6 mm. times.250 mm, 5 μm); the mobile phase is acetonitrile-phosphoric acid-0.05% trifluoroacetic acid (volume ratio is 250:1: 750); the detector is an ultraviolet detector; the detection wavelength is 210 nm; the column temperature is room temperature; the flow rate is 1.0 mL/min; the amount of sample was 20. mu.L.

Example 1

Formulation composition

Formulation components	Batch charging quantity (parts)
		Biotin crystal	10
Spicy sunflowerGlyceride esters (MCT)	25
		Stearic acid monoglyceride	1
Starch sodium octenyl succinate	30
		Sucrose	30.5
Glycerol	2
		Ascorbic acid sodium salt	1.5

S1, adding 10 parts of biotin crystals and 1 part of glycerol monostearate into 25 parts of MCT oil, stirring and dispersing uniformly, then grinding by a nano sand mill to enable the biotin crystals to reach nano-scale particles (less than 100nm), cooling to 20 ℃ by cooling water to prepare oil suspension I for later use.

S2, adding 30 parts of sodium starch octenylsuccinate, 30.5 parts of sucrose, 2 parts of glycerol and 1.5 parts of sodium ascorbate into 110 parts of pure water, heating to 50-60 ℃, stirring until the sodium starch is completely dissolved, and cooling to 20 ℃ by using cooling water to obtain a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, and then shearing and emulsifying for 15min at the temperature of 20 ℃ and the rotating speed of 10000rpm to ensure that the particle size D of oil drops₉₀Less than or equal to 2 microns, then homogenizing for 2 times under 50MPa, and detecting to obtain the oil drop grain diameter D of the high-pressure homogenized product₉₀Then, the viscosity of the emulsion was adjusted to 0.862 μm and 250 mPas, to obtain a biotin emulsion.

S4, spray drying the biotin emulsion, setting the inlet air temperature at 170 ℃, and dischargingThe wind temperature is 80 ℃, and the wind inlet quantity is 30m³The pressure of the spray tower is 0.4KPa, and the biotin microcapsule (particle diameter D) is obtained₉₀300 μm), 0.5 part of silica was mixed as an anticaking agent. The content of biotin in the microcapsules was 9.75% by HPLC.

Example 2

Formulation composition

Formulation components	Batch charging quantity (parts)
		Biotin crystal	5
Caprylic capric acid glyceride (MCT)	20
		Stearic acid monoglyceride	0.5
Starch sodium octenyl succinate	20
		Maltodextrin	50
Sorbitol	3
		Sodium hexametaphosphate	1.5

S1, adding 5 parts of biotin crystals and 0.5 part of stearic acid monoglyceride into 20 parts of MCT oil, stirring and dispersing uniformly, then grinding by a nano sand mill to enable the biotin crystals to reach nano-scale particles (less than 100nm), cooling to 15 ℃ by cooling water to prepare oil suspension I for later use.

S2, adding 20 parts of sodium starch octenylsuccinate, 50 parts of maltodextrin, 3 parts of sorbitol and 1.5 parts of sodium hexametaphosphate into 110 parts of pure water, heating to 50-60 ℃, stirring until the sodium starch hexametaphosphate is completely dissolved, and cooling to 15 ℃ by using cooling water to obtain a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, and then shearing and emulsifying at the temperature of 15 ℃ and the rotating speed of 11000rpm for 15min to ensure that the particle size D of oil drops₉₀Less than or equal to 2 microns, then homogenizing for 2 times under 60MPa, and detecting to obtain the oil drop grain diameter D of the high-pressure homogenized product₉₀Then, the viscosity of the emulsion was adjusted to 200 mPas to obtain a biotin emulsion, which was 0.756 μm.

S4, spray drying the biotin emulsion, setting the air inlet temperature at 165 ℃, the air outlet temperature at 75 ℃ and the air inlet amount at 30m³The pressure of the spray tower is 0.4KPa, and the biotin microcapsule (particle diameter D) is obtained₉₀350 μm) was mixed with 0.8 part of tricalcium phosphate as an anticaking agent. The content of biotin in the microcapsules was 4.85% as determined by high performance liquid chromatography.

Example 3

Formulation composition

Formulation components	Batch charging quantity (parts)
		Biotin crystal	15
Corn oil	30
		Lauric acid monoglyceride	1.5
Arabic gum	40
		Solid corn syrup	10
Mannitol	3
		Trisodium citrate	0.5

S1, adding 15 parts of biotin crystals and 1.5 parts of lauric acid monoglyceride into 30 parts of corn oil, stirring and dispersing uniformly, then grinding by a nano sand mill to enable biotin crystals to reach nano-scale particles (less than 100nm), cooling to 20 ℃ by cooling water to prepare oil suspension I for later use.

S2, adding 40 parts of Arabic gum, 10 parts of solid corn syrup, 3 parts of mannitol and 0.5 part of trisodium citrate into 110 parts of pure water, heating to 50-60 ℃, stirring until the materials are completely dissolved, and cooling to 20 ℃ with cooling water to obtain a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, and then shearing and emulsifying at the temperature of 20 ℃ and the rotating speed of 9000rpm for 20min to ensure that the oil drop diameter D₉₀Less than or equal to 2 microns, high-pressure homogenizing for 3 times under 55MPa, and detecting to obtain oil drop particle diameter D₉₀Then, the emulsion viscosity was adjusted to 220 mPas to 0.776 μm to obtain a biotin emulsion.

S4, spray drying the biotin emulsion, setting the air inlet temperature to be 160 ℃, the air outlet temperature to be 75 ℃ and the air inlet amount to be 30m³/h，Spraying tower pressure of 0.4KPa to obtain biotin microcapsule (particle diameter D)₉₀400 μm), 0.7 part of calcium silicate was mixed as an anticaking agent. The content of biotin in the microcapsules was 14.75% by HPLC.

Example 4

Formulation composition

Formulation components	Batch charging quantity (parts)
		Biotin crystal	10
Corn oil	25
		Lauric acid monoglyceride	1
Arabic gum	35
		Solid corn syrup	23
Hexapolyglycerol laurate	5
		Trisodium citrate	1

S1, adding 10 parts of biotin crystals and 1 part of lauric acid monoglyceride into 25 parts of corn oil, stirring and dispersing uniformly, then grinding by a nano sand mill to enable biotin crystals to reach nano-scale particles (less than 100nm), cooling to 18 ℃ by cooling water to obtain oil suspension I for later use.

S2, adding 35 parts of Arabic gum, 23 parts of solid corn syrup, 5 parts of hexaglycerol laurate and 1 part of trisodium citrate into 110 parts of pure water, heating to 50-60 ℃, stirring until the materials are completely dissolved, and cooling to 18 ℃ with cooling water to obtain a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, and then shearing and emulsifying for 12min at the temperature of 18 ℃ and the rotating speed of 10000rpm to ensure that the particle size D of oil drops₉₀Less than or equal to 2 microns, then homogenizing for 2 times under 50MPa, and detecting to obtain the oil drop grain diameter D of the high-pressure homogenized product₉₀Then, the viscosity of the emulsion was adjusted to 250 mPas to 0.766 μm, thereby obtaining a biotin emulsion.

S4, spray drying the biotin emulsion, setting the air inlet temperature at 165 ℃, the air outlet temperature at 80 ℃ and the air inlet amount at 30m³The pressure of the spray tower is 0.4KPa, and the biotin microcapsule (particle diameter D) is obtained₉₀500 μm), 0.5 part of silica was mixed as an anticaking agent. The content of biotin in the microcapsules was 9.87% by HPLC.

Example 5

Formulation composition

Formulation components	Batch charging quantity (parts)
		Biotin crystal	5
Soybean oil	20
		Lauric acid monoglyceride	0.5
Casein sodium salt	20
		Glucose	50
Glycerol	4
		Ethylenediaminetetraacetic acid sodium salt	0.5

S1, adding 5 parts of biotin crystals and 0.5 part of lauric acid monoglyceride into 20 parts of soybean oil, stirring and dispersing uniformly, then grinding by a nano sand mill to enable biotin crystals to reach nano-scale particles (less than 100nm), cooling to 15 ℃ by cooling water to prepare oil suspension I for later use.

S2, adding 20 parts of sodium caseinate, 50 parts of glucose, 4 parts of glycerol and 0.5 part of sodium ethylene diamine tetracetate into 100 parts of pure water, heating to 50-60 ℃, stirring until the sodium caseinate, the glucose, the glycerol and the sodium ethylene diamine tetracetate are completely dissolved, and cooling to 15 ℃ by using cooling water to obtain a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, and then shearing and emulsifying at the temperature of 15 ℃ and the rotating speed of 8500rpm for 25min to ensure that the oil drop diameter D₉₀Less than or equal to 2 microns, then homogenizing for 2 times under 60MPa, and detecting to obtain the oil drop grain diameter D of the high-pressure homogenized product₉₀Then, the emulsion viscosity was adjusted to 180 mPas to 0.665. mu.m, to obtain a biotin emulsion.

S4, spray drying the biotin emulsion, setting the air inlet temperature to be 160 ℃, the air outlet temperature to be 75 ℃ and the air inlet amount to be 30m³The pressure of the spray tower is 0.4KPa, and the biotin microcapsule (particle diameter D) is obtained₉₀450 μm), 0.7 part of microcrystalline cellulose was mixed as an anticaking agent. The content of biotin in the microcapsules was 4.82% as determined by high performance liquid chromatography.

Example 6

Formulation composition

Formulation components	Batch charging quantity (parts)
		Biotin crystal	15
Soybean oil	30
		Lauric acid monoglyceride	1.5
Casein sodium salt	40
		Glucose	10
Sorbitol	2
		Potassium tripolyphosphate	1.5

S1, adding 15 parts of biotin crystals and 1.5 parts of lauric acid monoglyceride into 30 parts of soybean oil, stirring and dispersing uniformly, then grinding by a nano sand mill to enable biotin crystals to reach nano-scale particles (less than 100nm), cooling to 16 ℃ by cooling water to prepare oil suspension I for later use.

S2, adding 40 parts of sodium caseinate, 10 parts of glucose, 2 parts of sorbitol and 1.5 parts of potassium tripolyphosphate into 100 parts of pure water, heating to 50-60 ℃, stirring until the sodium caseinate, the glucose, the sorbitol and the potassium tripolyphosphate are completely dissolved, and cooling to 16 ℃ with cooling water to obtain a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, and then shearing and emulsifying at the temperature of 16 ℃ and the rotating speed of 9000rpm for 15min to ensure that the oil drop diameter D₉₀Less than or equal to 2 microns, high-pressure homogenizing for 3 times under 55MPa, and detecting to obtain oil drop particle diameter D₉₀Then, the viscosity of the emulsion was adjusted to 190 mPas to 0.635. mu.m, thereby obtaining a biotin emulsion.

S4, spray drying the biotin emulsion, setting the air inlet temperature at 165 ℃, the air outlet temperature at 80 ℃ and the air inlet amount at 30m³The pressure of the spray tower is 0.4KPa, and the biotin microcapsule (particle diameter D) is obtained₉₀300 μm) was mixed with 0.8 part of tricalcium phosphate as an anticaking agent. The content of biotin in the microcapsules was 14.79% as determined by high performance liquid chromatography.

Example 7

Formulation composition

Formulation components	Batch charging quantity (parts)
		Biotin crystal	5
Rapeseed oil	20
		Sucrose ester (HLB value 5)	1
Whey protein	20
		Resistant dextrins	48
Glycerol	5
		Trisodium citrate	1

S1, adding 5 parts of biotin crystals and 1 part of sucrose ester (HLB value of 5) into 20 parts of rapeseed oil, stirring and dispersing uniformly, then grinding by a nano sand mill to enable the biotin crystals to reach nano-particles (less than 100nm), cooling to 12 ℃ by cooling water to prepare an oil suspension I for later use.

S2, adding 20 parts of whey protein, 48 parts of resistant dextrin, 5 parts of glycerol and 1 part of trisodium citrate into 100 parts of pure water, heating to 50-60 ℃, stirring until the materials are completely dissolved, cooling to 12 ℃ by using cooling water, and preparing a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, and then shearing and emulsifying at the temperature of 12 ℃ and the rotation speed of 8000rpm for 30min to ensure that the oil drop diameter D₉₀Less than or equal to 2 microns, then homogenizing for 3 times under 45MPa, and detecting to obtain the oil drop grain diameter D of the high-pressure homogenized product₉₀Then, the emulsion viscosity was adjusted to 185 mPas to 0.656. mu.m, thereby obtaining a biotin emulsion.

S4, spray drying the biotin emulsion, setting the air inlet temperature at 165 ℃, the air outlet temperature at 80 ℃ and the air inlet amount at 30m³The pressure of a spray tower is 0.4KPa, and the biotin microcapsule (particle diameter D) is prepared₉₀380 μm) was added, 0.8 part of tricalcium phosphate was added as an anticaking agent. Detecting the content of biotin in microcapsules by using a high performance liquid chromatographThe content was 4.88%.

Example 8

Formulation composition

Formulation components	Batch charging quantity (parts)
		Biotin crystal	10
Peanut oil	25
		Vitamin E polyethylene glycol 2000 succinate	0.5
Isolated soy protein	35
		Maltose	24
Hexapolyglycerol laurate	5
		Ascorbic acid	0.5

S1, adding 10 parts of biotin crystals and 0.5 part of vitamin E polyethylene glycol 2000 succinate into 25 parts of peanut oil, stirring and dispersing uniformly, then grinding by a nano sand mill to enable the biotin crystals to reach nano-particles (less than 100nm), cooling to 15 ℃ by cooling water to prepare an oil suspension I for later use.

S2, adding 35 parts of soybean protein isolate, 24 parts of maltose, 5 parts of hexaglycerol laurate and 0.5 part of ascorbic acid into 100 parts of pure water, heating to 50-60 ℃, stirring until the materials are completely dissolved, and cooling to 15 ℃ by using cooling water to obtain a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, and then shearing and emulsifying at the temperature of 15 ℃ and the rotation speed of 10000rpm for 15min to enable the oil drop particle size D₉₀Less than or equal to 2 microns, then homogenizing for 2 times under 55MPa, and detecting to obtain the oil drop grain diameter D of the high-pressure homogenized product₉₀The viscosity of the emulsion was adjusted to 0.750. mu.m, and then adjusted to 260 mPas, to obtain a biotin emulsion.

S4, spray drying the biotin emulsion, setting the air inlet temperature to be 170 ℃, the air outlet temperature to be 80 ℃ and the air inlet amount to be 30m³The pressure of the spray tower is 0.4KPa, and the biotin microcapsule (particle diameter D) is obtained₉₀430 μm), 0.5 parts of silica was mixed as an anticaking agent. The content of biotin in the microcapsules was 9.82% as determined by high performance liquid chromatography.

Example 9

Biotin microcapsules were prepared by the same method as in example 7 except that sucrose ester having HLB value of 5 was replaced with the same weight part of sucrose ester having HLB value of 7 under the same conditions as in example 7, to obtain biotin microcapsules having particle size D₉₀Is 460 μm. The content of biotin in the microcapsules was 4.90% by high performance liquid chromatography.

Comparative example 1

Biotin microcapsules were prepared according to the procedure of example 1, except that the biotin crystals and the stearic acid monoglyceride were mixed in MCT oil and then not ground by a nanomesher, the specific procedure was as follows:

s1, adding 10 parts of biotin crystals and 1 part of glycerol monostearate into 25 parts of MCT oil, stirring and dispersing uniformly, cooling to 20 ℃ by using cooling water to obtain an oil suspension I, wherein the crystal grain size of the biotin crystals is more than or equal to 50 microns for later use.

S2, adding 30 parts of sodium starch octenylsuccinate, 30.5 parts of sucrose, 2 parts of glycerol and 1.5 parts of sodium ascorbate into 110 parts of pure water, heating to 50-60 ℃, stirring until the sodium starch is completely dissolved, and cooling the water to 20 ℃ to obtain a water phase II for later use.

S3, mixing the oil suspension I and the water phase II, shearing and emulsifying at 20 ℃ and 10000rpm for 15min, homogenizing at 50MPa for 2 times, and detecting to obtain the oil drop diameter D of the high-pressure homogenized product₉₀And 55.5 μm (the actual particle diameter of the biotin crystal particles was measured), followed by adjusting the viscosity of the emulsion to 250 mPas to obtain a biotin emulsion.

S4, spray drying the biotin emulsion, setting the air inlet temperature to be 170 ℃, the air outlet temperature to be 80 ℃ and the air inlet amount to be 30m³The pressure of the spray tower is 0.4KPa, and the biotin microcapsule (particle diameter D) is obtained₉₀430 μm), 0.5 parts of silica was mixed as an anticaking agent. The content of biotin in the microcapsules was 9.05% as determined by high performance liquid chromatography.

Comparative example 2

A biotin batch was prepared as in example 1, except that caprylin caprin (MCT) was replaced with the same weight parts of sucrose and the raw materials were simply physically mixed to form the biotin batch, specifically: fully and uniformly mixing 10 parts of biotin crystals, 1 part of glycerol monostearate, 30 parts of sodium starch octenylsuccinate, 55.5 parts of sucrose, 2 parts of glycerol and 1.5 parts of sodium ascorbate to obtain a biotin mixed material, mixing 0.5 part of silicon dioxide as an anticaking agent, and detecting the content of biotin in the mixed material to be 8.45% by using a high performance liquid chromatograph.

Comparative example 3

A biotin batch was prepared as in example 1, except that caprylic capric acid glyceride (MCT) was replaced with the same weight parts of sucrose, and the biotin batch was prepared by completely dissolving the biotin crystals in hot water without embedding, and spray-drying after all the materials were dissolved together, specifically: adding 10 parts of biotin crystals into 110 parts of pure water, heating to 80-90 ℃ to completely dissolve the biotin crystals, and then adding 1 part of glycerol monostearate, 30 parts of sodium starch octenyl succinate and 2 parts of glycerol55.5 parts of cane sugar and 1.5 parts of sodium ascorbate are stirred and dissolved, and then spray drying is carried out, wherein the air inlet temperature is 170 ℃, the air outlet temperature is 80 ℃, and the air inlet amount is 30m³At 0.4KPa in spray tower pressure, biotin-containing mixture powder was obtained, and 0.5 part of silica was mixed as an anti-caking agent. The content of biotin in the powder was measured to be 9.45% by high performance liquid chromatography.

Comparative example 4

A biotin microcapsule was prepared by the method of example 1, except that the conditions for the nano-sand mill grinding in step S1 were controlled so that the particle diameter of the biotin crystal was 200 to 1000nm, and the other conditions were the same as in example 1, to obtain a biotin microcapsule having a particle diameter D₉₀And 390 μm. The content of biotin in the microcapsules was 9.15% as determined by high performance liquid chromatography.

Test example 1

The samples of examples 1-9 and comparative examples 1-4 were tested for solubility, specifically: the sample (1 g) was dissolved in 100mL of normal temperature water, and the time for the sample to completely settle in water (standing) and the dissolution of the aqueous solution were observed.

Table 1: table of solubility

Numbering	Solubility in water
		Example 1	The emulsion is uniform in 25s during dissolution
Example 2	The emulsion is uniform for 23s
		Example 3	The emulsion is uniform in 25s during dissolution
Example 4	The emulsion is uniform in 24s during dissolution
		Example 5	The emulsion is uniform in 24s during dissolution
Example 6	The emulsion is uniform for 23s
		Example 7	The emulsion is uniform for 21s
Example 8	26s when dissolved, the emulsion is uniform
		Example 9	The emulsion is uniform within 22s when dissolved
Comparative example 1	It can not be completely dissolved within 20min, the emulsion has obvious particles, and insoluble substances precipitate at the bottom
		Comparative example 2	It can not be completely dissolved within 20min, the emulsion has obvious particles, and insoluble substances precipitate at the bottom
Comparative example 3	It can not be completely dissolved within 20min, the emulsion has obvious particles, and insoluble substances precipitate at the bottom
		Comparative example 4	It can not be completely dissolved after 20min, and the emulsion has small amount of particles and bottomThere is insoluble substance precipitate

The results in table 1 show that the microencapsulated biotin crystals of examples 1 to 9 have nanocrystalline particles, fast dissolution and dispersion speed, and uniform emulsion after dissolution. The biotin crystals of comparative examples 1 and 4 have large particles and cannot be embedded without grinding or after grinding, and cannot be dissolved in water at normal temperature, and insoluble precipitates are the biotin crystals. Comparative examples 2 and 3 were simply physical mixing, were not microencapsulated, were insoluble in normal temperature water, and had large-particle crystalline precipitates of biotin.

Test example 2

The emulsion of test example 1 was measured for the particle size distribution of oil droplets or insoluble particles in the system using a malvern laser particle size analyzer 3000, and observed under a microscope, and the embedding of the emulsion in the granulation process of the microencapsulation technique was compared and judged, and the results are shown in table 2.

Table 2: oil droplet or insoluble particle distribution of multiple emulsions of each of the examples and comparative examples

The results in table 2 show that the biotin crystals in examples 1 to 9 were first nano-ground and then microencapsulated, the emulsion oil droplets were small in particle size, and each oil droplet contained a biotin microcrystal, which resulted in a good embedding effect. Comparative example 1, which was not ground, the biotin crystal particles were large and biotin could not be embedded, and large biotin crystal particles were tested. Comparative example 2 and comparative example 3 were simply physical mixing, without microencapsulation, and tested as large biotin crystal particles. Comparative example 4 the biotin crystals after milling were not fine enough, the particles were large, most were not encapsulated by oil droplets, the biotin was not completely embedded, and the particle size of the crystal particles was also tested.

Test example 3

Example 1, example 3, example 5 and exampleThe samples of example 7 and comparative examples 1 to 4 were placed in sealed, colorless and transparent vials, respectively, and placed in ultraviolet aging test chambers (irradiance of 2W/m)²25 ℃) and oxygen filling (80 ℃) for 30 days, sampling respectively for 0 day, 10 days, 20 days and 30 days, detecting the content of biotin in each sample by using a high performance liquid chromatograph, and examining the influence of ultraviolet irradiation and oxygen filling high temperature conditions on the content (stability) of biotin in the sample, wherein the results are shown in table 3.

Table 3: influence of ultraviolet irradiation and oxygenation high-temperature conditions on stability of biotin in sample

From the results in table 3, it can be seen that the biotin microcapsules obtained by the method provided by the present invention are placed under the conditions of ultraviolet irradiation and high temperature of oxygen, and accelerated aging is performed, sampling tests are performed for 0 day, 10 days, 20 days, and 30 days, and the sample microcapsules of examples 1, 3, 5, and 7 have substantially no significant drop in the biotin content, while the sample microcapsules of comparative examples 1 to 4 have significantly large drops, which indicates that the biotin microcapsules obtained by the method provided by the present invention have good stability and can prolong the storage time of the biotin active ingredients.

Test example 4

In order to verify that the biotin microcapsules prepared by the invention have unique acid resistance, the samples of the above examples 1, 3, 5 and 7 and comparative examples 1 to 4 are subjected to a stability experiment simulating gastric juice in a strong acid environment. Preparing a buffer solution with the pH value of 1.0, dissolving and dispersing 5g of a sample in 200mL of the buffer solution in a closed container with magnetic stirring, keeping the temperature of the solution at 37 +/-0.5 ℃, starting the magnetic stirring (rotating speed of 100rpm), immediately starting timing, starting a simulated gastric juice digestion experiment, wherein the simulation time is 3h, sampling and detecting at 0, 30min, 60min, 90min, 120min and 180min respectively, and examining the retention rate of biotin content (%), and the results are shown in Table 4.

Table 4: simulating the destruction of the strong acid environment of the stomach to the biotin in the sample

As can be seen from the results in Table 4, when the experiment is carried out in a simulated strong acid environment in the stomach, the retention rate of the biotin content of the samples of example 1, example 3, example 5 and example 7 is still high at 3h, and the embedded biotin resists the corrosion of acid substances in the stomach, so that the loss of the biotin content is reduced. And in comparative examples 1-4, the biotin microcapsule prepared by the method is not used or is not completely used, the loss of biotin content of the obtained sample is large under the action of strong acid in the stomach, the sample cannot effectively reach the small intestine, and the bioavailability is reduced.

In conclusion, the biotin microcapsule obtained by the method provided by the invention has better acid resistance, is not damaged in the stomach, and can effectively reach the intestines to be absorbed.

Test example 5

The results of the investigation of the washability of the samples of examples 1, 3, 5 and 7 and comparative examples 1 to 4 at different pH values are shown in Table 5.

Table 5: the reconstitution behavior of the biotin microcapsules under different pH values

From the results of table 5, it can be seen that the biotin microcapsule powders of examples 1, 3, 5 and 7 have excellent acid and alkali resistance and can be normally reconstituted in various pH ranges. Comparative examples 1 to 4 were not completely soluble at each pH, the emulsion had significant insoluble particles, bottom precipitates, and poor reconstitution properties.

Although embodiments of the present invention have been shown and described above, it is understood that the above embodiments are exemplary and should not be construed as limiting the present invention, and that variations, modifications, substitutions and alterations can be made in the above embodiments by those of ordinary skill in the art without departing from the principle and spirit of the present invention.