Product containing flexible glucan, preparation method, composition, and use

Abstract

The present invention relates to the technical field of cosmetics, and in particular to a product containing flexible glucan. The product can be prepared by using a high-pressure micro-jet to treat a material containing glucan. In the product containing flexible glucan of the present invention, the flexible glucan has special intramolecular and intermolecular arrangements, achieves excellent wound-healing-promoting and anti-inflammatory effects, when added to a cosmetic, shows a prominent skin-repair-promoting effect, can effectively improve the effect after medical aesthetic procedures such as fractional laser, chemical peel, and microneedling, and can also promote the repair of sensitive skin. The preparation method for the product containing flexible glucan of the present invention allows for adjustment of the intramolecular and intermolecular arrangements of the glucan and enhances the stability of the product at room temperature while improving the flexibility of the product, so that a large amount of the product can be added to a cosmetic.

Description

Product containing flexible glucan, preparation method, composition and application TECHNICAL FIELD

[0001] The present application relates to the technical field of skin care products, in particular to a product containing flexible glucan and its preparation and application. BACKGROUND

[0002] Glucan will cause intramolecular and intermolecular bonding due to its polyhydroxy structure and long straight chain during storage, which will further cause aggregation and agglomeration of glucan molecules. On the one hand, the biological activity is reduced due to the closed active sites caused by agglomeration, and on the other hand, the product is unstable due to the agglomeration between molecules.

[0003] In related technologies, in order to improve the stability of glucan, means such as changing functional groups and removing macromolecular glucan in the preparation process are adopted. These means essentially change the composition of the glucan product, and also make the preparation process more complex and increase the production cost.

[0004] Therefore, the present application is proposed. SUMMARY

[0005] An object of the present application is to provide a product containing flexible glucan, which can solve the problems of poor efficacy and poor stability of such products during long-term storage.

[0006] Another object of the present application is to provide a preparation method of a product containing flexible glucan.

[0007] Still another object of the present application is to provide a composition containing the product containing flexible glucan described above.

[0008] Still another object of the present application is to provide an application of the product containing flexible glucan / composition described above in the preparation of medical products and / or cosmetics.

[0009] In order to achieve the above objects of the present application, the present application provides a product containing flexible glucan, wherein the flexible glucan molecules in the product containing flexible glucan have a globular chain structure, and the molecular conformation parameter is 0.5-0.8.

[0010] The product containing flexible glucan of the present application has a special intramolecular and intermolecular arrangement of glucan, has excellent effects of promoting wound healing and anti-inflammatory, and when added to cosmetics, has outstanding effects of promoting skin repair, can effectively improve the healing effects of medical beauty projects such as fractional laser, acid brushing and microneedle, and can promote the repair of sensitive skin.

[0011] Another aspect of the present application provides a method for preparing a product containing flexible glucan, comprising treating a glucan-containing material with high-pressure microfluidization.

[0012] The method for preparing a product containing flexible glucan of the present application can adjust the arrangement within and between glucan molecules, increase the flexibility of the product while increasing its room temperature stability, so that it can be added in large amounts in cosmetics.

[0013] In one embodiment of the present application, the glucan comprises beta-glucan.

[0014] In one embodiment of the present application, the glucan comprises cereal beta-glucan.

[0015] In one embodiment of the present application, the glucan comprises one or more of oat beta-glucan, barley beta-glucan, and barley beta-glucan.

[0016] In one embodiment of the present application, the glucan-containing material comprises a glucan-containing extract.

[0017] In one embodiment of the present application, during the treatment of the glucan-containing material with high-pressure microfluidization, the mass ratio of the glucan-containing material to the fluid is 1:(12-70), and further preferably 1:14.

[0018] In one embodiment of the present application, the fluid comprises water.

[0019] In one embodiment of the present application, the high-pressure microfluidization pressure is 80-140 MPa gauge, and further preferably 120 MPa gauge.

[0020] In one embodiment of the present application, the high-pressure microfluidization is performed 1-5 times, and further preferably 2 times.

[0021] In one embodiment of the present application, the high-pressure microfluidization feed temperature is 0-45°C; and further preferably 25°C.

[0022] In one embodiment of the present application, the glucan-containing material is heated and dissolved before being treated with high-pressure microfluidization.

[0023] In one embodiment of the present application, the heating and dissolving temperature is 70-90°C; and further preferably, the heating and dissolving temperature is 80-85°C.

[0024] In one embodiment of the present application, the method further comprises at least one of decolorization, desalination, and deproteinization.

[0025] In one embodiment of the present application, the number of decoloring is 1-3 times, preferably 2 times.

[0026] In one embodiment of the present application, the total amount of decoloring agent added is 2%-5% of the total mass of the material to be decolored, further preferably 2-3%.

[0027] In one embodiment of the present application, the decoloring agent comprises activated carbon.

[0028] In one embodiment of the present application, the time for each decoloring is 30-60 min.

[0029] In one embodiment of the present application, the temperature for each decoloring is 75-85℃.

[0030] In one embodiment of the present application, the number of decoloring is 2 times, the first time adding 3% of the total mass of the material to be decolored, and the second time adding 2% of the total mass of the material to be decolored, and the decoloring is carried out at 80-85℃ for 30 min.

[0031] In one embodiment of the present application, the desalination is carried out using cation and anion exchange resins.

[0032] In one embodiment of the present application, the flow rate of the material during desalination is 1-3 t / h, further preferably, the flow rate of the material during desalination is 2-3 t / h.

[0033] In one embodiment of the present application, the obtained product is filtered and reconstituted to obtain a product containing flexible glucan.

[0034] In one embodiment of the present application, one or more of hexylene glycol, pentylene glycol, glycerol and PEHG is used as a reconstitution solvent.

[0035] In one embodiment of the present application, sterilization is carried out after reconstitution.

[0036] Another aspect of the present application provides a composition containing flexible glucan, which comprises the above-mentioned product containing flexible glucan, and / or the product containing flexible glucan prepared by the above-mentioned method for preparing a product containing flexible glucan.

[0037] Another aspect of the present application provides the use of the above-mentioned product containing flexible glucan, the product containing flexible glucan prepared by the above-mentioned method for preparing a product containing flexible glucan, or the above-mentioned composition containing flexible glucan in the preparation of a pharmaceutical product and / or a cosmetic product.

[0038] Compared with the prior art, the present application has the following beneficial effects:

[0039] The product containing flexible glucan of the present application has excellent effects of promoting wound healing and anti-inflammation, and when added to cosmetics, the product has outstanding effects of promoting skin repair and can effectively improve the healing effects of medical and beauty projects such as fractional laser, acid brushing and microneedle, and can promote the repair of sensitive skin.

[0040] The preparation method of the product containing flexible glucan of the present application can adjust the arrangement form of glucan molecules and increase the flexibility and room temperature stability of the product, so that the product can be added in large amounts in cosmetics. BRIEF DESCRIPTION OF DRAWINGS

[0041] In order to more clearly illustrate the specific embodiments of the present application or the technical solutions in the prior art, the drawings needed in the following specific embodiment or prior art description will be briefly introduced. Obviously, the drawings in the following description are some embodiments of the present application, and other drawings can also be obtained by those skilled in the art without creative labor.

[0042] Figure 1 is a scanning electron microscope image of the flexible glucan sample of Example 3 of the present application, wherein Figure 1a and Figure 1b are scanning electron microscope images of the flexible glucan sample of Example 3 of the present application at different scales;

[0043] Figure 2 is a scanning electron microscope image of the flexible glucan sample of Example 7 of the present application, wherein Figure 2a and Figure 2b are scanning electron microscope images of the flexible glucan sample of Example 7 of the present application at different scales;

[0044] Figure 3 is a scanning electron microscope image of the glucan sample of Comparative Example 1 of the present application, wherein Figure 3a and Figure 3b are scanning electron microscope images of the glucan sample of Comparative Example 1 of the present application at different scales;

[0045] Figure 4 is a scanning electron microscope image of the glucan sample of Comparative Example 2 of the present application, wherein Figure 4a and Figure 4b are scanning electron microscope images of the glucan sample of Comparative Example 2 of the present application at different scales;

[0046] Figure 5 is a scanning electron microscope image of the glucan sample of Comparative Example 3 of the present application, wherein Figure 5a and Figure 5b are scanning electron microscope images of the glucan sample of Comparative Example 3 of the present application at different scales;

[0047] Figure 6 is a scanning electron microscope image of the glucan sample of Comparative Example 4 of the present application, wherein Figure 6a and Figure 6b are scanning electron microscope images of the glucan sample of Comparative Example 4 of the present application at different scales;

[0048] Figure 7 is an atomic force microscope image of the flexible glucan sample of Example 3 of the present application, wherein Figure 7a and Figure 7b are atomic force microscope images of the flexible glucan sample of Example 3 of the present application at different scales, respectively;

[0049] Figure 8 is an atomic force microscope image of the glucan sample of Comparative Example 2 of the present application, wherein Figure 8a and Figure 8b are atomic force microscope images of the glucan sample of Comparative Example 2 of the present application at different scales, respectively;

[0050] Figure 9 is a bright field image (100X) of a scratch wound healing of keratinocytes (HaCaT) in the specific embodiment of the present application;

[0051] Figure 10 is a graph of comparison data of the scratch wound healing rate of the flexible glucan sample in the specific embodiment of the present application;

[0052] Figure 11 is a graph of comparison data of the anti-inflammatory efficacy characterization of the flexible glucan sample in the specific embodiment of the present application. DETAILED DESCRIPTION

[0053] The technical solutions of the present application will be described clearly and completely in combination with the drawings and specific embodiments below, but those skilled in the art will understand that the following described embodiments are part of the embodiments of the present application, not all the embodiments, and are only used to illustrate the present application, and should not be regarded as limiting the scope of the present application. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without making creative efforts are within the scope of protection of the present application. The specific conditions are not specified in the embodiments, and are carried out according to the conventional conditions or the conditions recommended by the manufacturer. The reagents or instruments used are not specified by the manufacturer, and are conventional products that can be purchased on the market.

[0054] In one aspect, the present application provides a product containing flexible glucan, wherein the flexible glucan molecules in the product containing flexible glucan have a globular chain structure, and the molecular conformation parameter is 0.5-0.8.

[0055] The product containing flexible glucan, wherein the glucan has a special intramolecular and intermolecular arrangement, has excellent effects of promoting wound healing and anti-inflammation, and when added to cosmetics, has outstanding effects of promoting skin repair, can effectively improve the postoperative effects of medical and beauty projects such as fractional laser, acid brushing, and microneedle, and can promote the repair of sensitive skin.

[0056] In another aspect, the present application provides a preparation method of a product containing flexible glucan, comprising treating a material containing glucan by high-pressure microjet.

[0057] High pressure micro jet can have a series of comprehensive effects on fluid mixture, such as strong shearing and high speed impact, and is often used for destroying / breaking materials and homogenizing materials. Mainly, high pressure (generally above 80 Mpa) is generated by a hydraulic pump, so that the fluid in the impact chamber is dispersed into two or more streams, and strong high-speed impact is carried out in a small flow channel space. In the process of impact, most of the energy is converted instantaneously, a huge pressure drop is generated, so that the liquid particles are highly broken. The present application creatively uses high pressure micro jet to modify the flexibility of dextran, and various high pressure micro jet devices can be selected for operation.

[0058] This method can adjust the arrangement form of dextran molecules and increase the flexibility and room temperature stability of the product, so that the product can be added in large amounts in cosmetics.

[0059] In one embodiment of the present application, the dextran includes beta-glucan, which is a natural product, a polysaccharide connected by glucose molecules, and exists in various plants and fungi. Beta-glucan has many important biological activities and pharmacological effects, and is widely used in the fields of medicine, health care products, food, etc.

[0060] In one embodiment of the present application, the dextran suitable for the present application can come from different sources, and the dextran includes cereal beta-glucan. All cereal beta-glucans are unbranched linear non-starch polysaccharides connected by beta-1, 3 glycosidic bonds and beta-1, 4 glycosidic bonds, so different cereal beta-glucans have similar properties, that is, they will all be unstable due to hydrogen bonding.

[0061] In one embodiment of the present application, the dextran includes one or more of oat beta-glucan, barley beta-glucan and barley beta-glucan.

[0062] In one embodiment of the present application, the material containing dextran includes an extract containing dextran. The dextran can be obtained from various sources by conventional extraction, extraction, etc., or can be directly purchased.

[0063] In one embodiment of the present application, during the process of treating the material containing dextran by using high pressure micro jet, the mass ratio of the material containing dextran to the fluid is 1: (12-70), and is further preferably 1:14.

[0064] The material containing dextran with different concentrations can be treated by high-pressure microfluidization according to the actual equipment and the target product concentration requirements. The flexible modification requirements of dextran can be met in the mass ratio range of the material containing dextran and the fluid, for example, the mass ratio of the material containing dextran and the fluid can be 1:12, 1:12.5, 1:14, 1:18, 1:20, 1:26, 1:34, 1:37, 1:42, 1:50, 1:53, 1:62, 1:69, 1:70, etc.

[0065] In one embodiment of the present application, the fluid comprises water.

[0066] In one embodiment of the present application, the high-pressure microfluidization pressure is 80-140 MPa, and further preferably 120 MPa.

[0067] In different embodiments, the high-pressure microfluidization pressure can be 80 MPa, 85 MPa, 90 MPa, 102 MPa, 114 MPa, 120 MPa, 131 MPa, 140 MPa, etc.

[0068] In one embodiment of the present application, the high-pressure microfluidization frequency is 1-6 times, and further preferably 2 times.

[0069] In different embodiments, the high-pressure microfluidization frequency can be 1 time, 2 times, 3 times, 4 times, 5 times, 6 times.

[0070] In one embodiment of the present application, the high-pressure microfluidization feeding temperature is 0-45℃; further preferably 25℃.

[0071] The feeding temperature of the high-pressure microfluidization can be adjusted according to the use requirements of the relevant equipment, or the actual production line design and other needs. There is no strict temperature control requirement, and it can be fed at room temperature. The flexible modification requirements of dextran can be met in the above high-pressure microfluidization feeding temperature range. For example, in different embodiments, the high-pressure microfluidization feeding temperature can be 0℃, 4℃, 8℃, 15℃, 20℃, 26℃, 32℃, 37℃, 41℃, 45℃, etc. If the specific feeding temperature is not mentioned in the embodiment of the present application, the above material temperature or room temperature feeding can be used.

[0072] In one embodiment of the present application, the material containing dextran is heated and dissolved before high-pressure microfluidization treatment. For general dextran materials, heating is a convenient operation to dissolve them, which facilitates the subsequent high-pressure microfluidization treatment to fully play its role.

[0073] In an embodiment of the present application, the heating dissolution temperature is 70-90°C; further preferably, the heating dissolution temperature is 80-85°C.

[0074] The heating can improve the dissolution efficiency of dextran, and the dextran can be dissolved at a suitable temperature without strict temperature requirements. In the above dissolution temperature range, the flexible modification requirements of dextran can be met. For example, in different embodiments, the heating dissolution temperature can be exemplarily 70-73°C, 74-76°C, 78-80°C, 81-83°C, 88-90°C, etc.

[0075] In an embodiment of the present application, the preparation method further comprises at least one of decolorization, desalination, and protein removal. Other corresponding treatment methods can be selected according to actual needs, such as the needs of subsequent formula applications.

[0076] In an embodiment of the present application, the number of decolorization is 1-3 times, preferably 2 times.

[0077] In an embodiment of the present application, the total addition amount of the decolorizing agent in the decolorization is 2%-5% of the total mass of the material to be decolorized, and further preferably 2-3%.

[0078] The total addition amount of the decolorizing agent can be determined according to the colority of the obtained product and the decolorization requirements. The use of conventional decolorizing agents does not affect the flexible properties of the dextran that has been processed. For example, in different embodiments, the total addition amount of the decolorizing agent in the decolorization can be exemplarily 2%, 2.1%, 2.4%, 2.9%, 3.15%, 3.6%, 4.2%, 4.74%, 5% of the total mass of the material to be decolorized, etc.

[0079] In an embodiment of the present application, the decolorizing agent comprises activated carbon.

[0080] In an embodiment of the present application, the time for each decolorization is 30-60 min.

[0081] For example, in different embodiments, the time for each decolorization can be exemplarily 30 min, 34 min, 39 min, 43 min, 46 min, 52 min, 57 min, 60 min, etc.

[0082] In an embodiment of the present application, the temperature for each decolorization is 75-85°C.

[0083] The decolorization temperature can be set according to the equipment, production line, decolorization efficiency and other requirements, and the decolorization temperature does not affect the flexibility of the dextran that has been processed. For example, in different embodiments, the temperature of each decolorization can be 75-77℃, 76-78℃, 78-80℃, 78.5-81℃, 83-85℃, etc.

[0084] In one specific embodiment of the present application, the number of decolorization is 2, the first time adding 3% of the total mass of the decolorization agent, and the second time adding 2% of the total mass of the decolorization agent activated carbon, and the temperature is 80-85℃ for 30 minutes.

[0085] In one specific embodiment of the present application, cation and anion exchange resins are used for desalination.

[0086] In one specific embodiment of the present application, the flow rate of the material during desalination is 1-3t / h, and further preferably, the flow rate of the material during desalination is 2-3t / h.

[0087] The flow rate of the material during desalination can be set according to the equipment, production line, desalination efficiency and other requirements, and the flow rate of the material does not affect the flexibility of the dextran that has been processed. For example, in different embodiments, the flow rate of the material during desalination can be 1-1.3t / h, 1.2-1.6t / h, 1.9-2.4t / h, 2-2.5t / h, 2.3-2.7t / h, 2.8-3t / h, etc.

[0088] By isoelectric point and combined with heating treatment, trace amounts of protein contained in the sample can be destroyed, denatured and precipitated, and then removed by subsequent filtration steps, thereby improving the appearance and stability of the product. The present application can also use other protein removal processes as alternatives, such as Sevage method, TCA method and protease method, etc.

[0089] In one specific embodiment of the present application, the obtained product is filtered and compounded to obtain a product containing flexible dextran.

[0090] In one specific embodiment of the present application, one or more of hexylene glycol, pentylene glycol, glycerol and PEHG are used as compounding solvents for compounding.

[0091] In one specific embodiment of the present application, the amount of the compounding solvent can be adjusted as needed. In one preferred specific embodiment of the present application, the obtained flexible dextran is added to glycerol, hexylene glycol and pentylene glycol, and the content of glycerol in the obtained product is 10% of the total mass, the content of hexylene glycol is 2% of the total mass, and the content of pentylene glycol is 2% of the total mass.

[0092] In one embodiment of the present application, the complexing is followed by sterilization.

[0093] In another aspect of the present application, there is provided a composition containing flexible glucan, containing the above-mentioned product containing flexible glucan, and / or containing the product containing flexible glucan prepared by the above-mentioned method for preparing a product containing flexible glucan.

[0094] In another aspect of the present application, there is provided the use of the above-mentioned product containing flexible glucan, the product containing flexible glucan prepared by the above-mentioned method for preparing a product containing flexible glucan, or the above-mentioned composition containing flexible glucan in the preparation of a pharmaceutical product and / or a cosmetic product.

[0095] Example 1

[0096] The flexible oat β-glucan is prepared by the following specific steps:

[0097] 7 kg of oat β-glucan powder is dissolved in 98 kg of water (solid-liquid ratio of 1:14), heated to 83°C for 30 min, and cooled to 25°C to obtain solution A;

[0098] Solution A is subjected to high-pressure microjet at a pressure of 120 MPa (gauge pressure), and is circulated for 2 times to obtain 60 kg of solution B;

[0099] Solution B is diluted with deionized water according to the mass ratio of solution B:water = 1:2 to obtain solution C;

[0100] Solution C is stirred and heated to 75°C, and 3% of activated carbon based on the mass of solution C is added, and then heated to 83°C for 30 min, and then settled for 12-18 h, and then the supernatant is extracted to obtain solution D;

[0101] Solution D is heated to 75°C, and 2% of activated carbon based on the mass of solution D is added, and then heated to 84.8°C for 30 min, and then cooled to 62°C, and then filtered to obtain solution E;

[0102] Solution E is subjected to desalination by passing through a cation-anion chromatographic column at a flow rate of 3.0 TPH, and then the pH is adjusted to 4.64 to obtain solution F;

[0103] Solution F is added with 1% of activated carbon based on the mass of solution F, and then heated to 92°C for 20 min, and then filtered to obtain flexible oat β-glucan;

[0104] The flexible oat β-glucan is added with glycerol, hexylene glycol and pentylene glycol, and the obtained product contains 10% of glycerol based on the total mass, 2% of hexylene glycol based on the total mass, and 2% of pentylene glycol based on the total mass, and then sterilized at 100°C for 30 min.

[0105] Example 2

[0106] Preparation of flexible oat β-glucan, the specific steps are as follows:

[0107] The 20 kg of oat β-glucan aqueous solution with a mass fraction of 1% obtained on the market (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd., which also contains glycerol with a mass fraction of 12% and PEHG with a mass fraction of 1%) is subjected to high-pressure microfluidization at a pressure of 120 MPa of gage pressure, and is circulated 1 time to obtain 20 kg of flexible oat β-glucan.

[0108] Example 3

[0109] Preparation of flexible oat β-glucan, the specific steps are as follows:

[0110] The 20 kg of oat β-glucan aqueous solution with a mass fraction of 1% obtained on the market (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd., which also contains glycerol with a mass fraction of 12% and PEHG with a mass fraction of 1%) is subjected to high-pressure microfluidization at a pressure of 120 MPa of gage pressure, and is circulated 2 times to obtain 20 kg of flexible oat β-glucan.

[0111] Example 4

[0112] Preparation of flexible oat β-glucan, the specific steps are as follows:

[0113] The 20 kg of oat β-glucan aqueous solution with a mass fraction of 1% obtained on the market (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd., which also contains glycerol with a mass fraction of 12% and PEHG with a mass fraction of 1%) is subjected to high-pressure microfluidization at a pressure of 120 MPa of gage pressure, and is circulated 3 times to obtain 20 kg of flexible oat β-glucan.

[0114] Example 5

[0115] Preparation of flexible oat β-glucan, the specific steps are as follows:

[0116] The 20 kg of oat β-glucan aqueous solution with a mass fraction of 1% obtained on the market (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd., which also contains glycerol with a mass fraction of 12% and PEHG with a mass fraction of 1%) is subjected to high-pressure microfluidization at a pressure of 120 MPa of gage pressure, and is circulated 5 times to obtain 20 kg of flexible oat β-glucan.

[0117] Example 6

[0118] Preparation of flexible oat β-glucan, the specific steps are as follows:

[0119] The 20 kg of the obtained oat β-glucan aqueous solution with a mass fraction of 1% purchased on the market (purchased from Beijing Dongfangmiosen Biotechnology Co., Ltd., which also contains glycerol with a mass fraction of 12% and PEHG with a mass fraction of 1%) is subjected to high-pressure microjet at a pressure of 120 MPa, and is circulated for 5 times to obtain 20 kg of flexible oat β-glucan.

[0120] Example 7

[0121] The flexible oat β-glucan is prepared, and the specific steps are as follows:

[0122] The 20 kg of the obtained oat β-glucan aqueous solution with a mass fraction of 1% purchased on the market (purchased from Beijing Dongfangmiosen Biotechnology Co., Ltd., which also contains glycerol with a mass fraction of 12% and PEHG with a mass fraction of 1%) is subjected to high-pressure microjet at a pressure of 120 MPa, and is circulated for 6 times to obtain 20 kg of flexible oat β-glucan.

[0123] Example 8

[0124] The flexible oat β-glucan is prepared, and the specific steps are as follows:

[0125] 5.1 kg of oat β-glucan powder (purchased from Beijing Dongfangmiosen Biotechnology Co., Ltd.) is dissolved in 71.4 kg of water, heated to 85°C for 30 min, and cooled to 25°C to obtain solution A;

[0126] Solution A is subjected to high-pressure microjet at a pressure of 120 MPa, and is circulated for 2 times to obtain 60 kg of solution B;

[0127] Solution B is diluted with deionized water according to a mass ratio of solution B: water = 1:2 to obtain solution C;

[0128] Then, 5% of activated carbon based on the mass of solution C is added, and decolorization is performed at 82°C for 30 min, and then cooling and filtration are performed to obtain solution D;

[0129] Solution D is subjected to desalination through an anion-cation chromatographic column at a flow rate of 3.0 TPH, and then the pH is adjusted to 5 to obtain solution E;

[0130] Solution E is continuously cooled to 30°C and filtered to obtain flexible oat β-glucan;

[0131] The flexible oat β-glucan is added with glycerol, hexylene glycol and pentylene glycol, and the obtained product contains glycerol with a mass fraction of 10% of the total mass, hexylene glycol with a mass fraction of 2% of the total mass, and pentylene glycol with a mass fraction of 2% of the total mass, and then sterilization is performed at 100°C for 30 min.

[0132] Example 9

[0133] The flexible oat β-glucan is prepared, and the specific steps are as follows:

[0134] 5.1 kg oat beta-glucan powder (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd.) was dissolved in 71.4 kg water, heated to 85°C for 30 min, and cooled to 25°C to obtain solution A;

[0135] Solution A was subjected to high-pressure micro-jet at a pressure of 120 MPa (gauge pressure), and circulated for 2 times to obtain 60 kg of solution B;

[0136] Solution B was diluted with deionized water according to the mass ratio of solution B: water = 1:2 to obtain solution C;

[0137] Then 3% of the mass of solution C was added to solution C to add activated carbon, and decolorization was carried out at 82°C for 30 min, and then cooled and filtered to obtain solution D;

[0138] Solution D was added with 2% of the mass of solution D to add activated carbon, and decolorization was carried out at 85°C for 30 min, and then cooled and filtered to obtain solution E;

[0139] Solution E was subjected to desalination through anion and cation chromatographic columns at a flow rate of 3.0 TPH, and then the pH was adjusted to 5 to obtain solution F;

[0140] Solution F was further cooled to 30°C and filtered to obtain flexible oat beta-glucan;

[0141] The flexible oat beta-glucan was added to glycerol, hexylene glycol and pentylene glycol, and the obtained product had a glycerol content of 10% of the total mass, a hexylene glycol content of 2% of the total mass, and a pentylene glycol content of 2% of the total mass, and then sterilized at 100°C for 30 min.

[0142] Example 10

[0143] Flexible oat beta-glucan was prepared from oat beta-glucan powder, and the specific steps were as follows:

[0144] Oat beta-glucan powder (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd.) was dissolved in water according to the mass ratio of oat beta-glucan powder: water = 1:12, heated to 83°C for 30 min, and cooled to 25°C to obtain solution A;

[0145] Solution A was subjected to high-pressure micro-jet at a pressure of 80 MPa (gauge pressure), and circulated for 2 times to obtain solution B;

[0146] Solution B was diluted with water to a mass fraction of 1% of oat beta-glucan to obtain solution C;

[0147] Solution C was stirred and heated to 75°C, 3% of the mass of solution C was added to add activated carbon, and heating was continued to 83°C for 30 min, and after 12-18 h of settling, the supernatant was extracted to obtain solution D;

[0148] Solution D was heated to 75°C, 2% activated carbon of the mass of solution D was added, and the temperature was kept at 84.8°C for 30 min, and then cooled to 62°C, and filtration was performed to obtain solution E;

[0149] Solution E was desalted by passing through an anion-cation chromatographic column at a flow rate of 3.0 TPH, and then the pH was adjusted to 4.64 to obtain solution F;

[0150] Activated carbon of 1% of the mass of solution F was added to solution F, and the temperature was then increased to 92°C and kept for 20 min, and then filtration was performed to obtain flexible oat β-glucan;

[0151] The flexible oat β-glucan was added to glycerol, hexylene glycol, and pentylene glycol, and the resulting product had a glycerol content of 10% of the total mass, a hexylene glycol content of 2% of the total mass, and a pentylene glycol content of 2% of the total mass, and then sterilized at 100°C for 30 min.

[0152] Example 11

[0153] Flexible oat β-glucan was prepared from oat β-glucan powder, and the specific steps were as follows:

[0154] Oat β-glucan powder (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd.) was dissolved in water at a mass ratio of oat β-glucan powder: water = 1:20, heated to 83°C and kept for 30 min, and then cooled to 25°C to obtain solution A;

[0155] Solution A was subjected to high-pressure micro-jet at a pressure of 100 MPa, and circulated for 2 times to obtain solution B;

[0156] Solution B was diluted with water to a mass fraction of 1% of oat β-glucan to obtain solution C;

[0157] Solution C was stirred and heated to 75°C, 3% activated carbon of the mass of solution C was added, and the temperature was then increased to 83°C and kept for 30 min, and after settling for 12-18 h, the supernatant was extracted to obtain solution D;

[0158] Solution D was heated to 75°C, 2% activated carbon of the mass of solution D was added, and the temperature was kept at 84.8°C for 30 min, and then cooled to 62°C, and filtration was performed to obtain solution E;

[0159] Solution E was desalted by passing through an anion-cation chromatographic column at a flow rate of 3.0 TPH, and then the pH was adjusted to 4.64 to obtain solution F;

[0160] Activated carbon of 1% of the mass of solution F was added to solution F, and the temperature was then increased to 92°C and kept for 20 min, and then filtration was performed to obtain flexible oat β-glucan;

[0161] The flexible oat beta-glucan is added into glycerol, hexylene glycol and pentylene glycol, the glycerol content in the obtained product is 10% of the total mass, the hexylene glycol content is 2% of the total mass, and the pentylene glycol content is 2% of the total mass, and then sterilized at 100℃ for 30 min.

[0162] Comparative Example 1

[0163] The oat beta-glucan is prepared from oat beta-glucan powder, and the specific steps are as follows:

[0164] 7 kg of oat beta-glucan powder (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd.) is dissolved in 98 kg of water (solid-liquid ratio is 1:14), heated to 83℃ and kept for 30 min, and cooled to 25℃ to obtain solution A;

[0165] Solution A is diluted with deionized water according to the mass ratio of solution A:water = 1:2 to obtain solution B;

[0166] Solution B is stirred and heated to 75℃, and 3% of activated carbon based on the mass of solution B is added, and then heated to 83℃ and kept for 30 min, and then settled for 12-18 h to obtain solution C;

[0167] Solution C is heated to 75℃, 2% of activated carbon based on the mass of solution C is added, and then kept at 84.8℃ for 30 min, and then cooled to 62℃, and then filtered to obtain solution D;

[0168] Solution D is desalted through an anion-cation chromatographic column at a flow rate of 3.0 TPH, and then the pH is adjusted to 4.64 to obtain solution E;

[0169] Solution E is added with 1% of activated carbon based on the mass of solution E, and then heated to 92℃ and kept for 20 min, and then filtered to obtain oat beta-glucan;

[0170] The flexible oat beta-glucan is added into glycerol, hexylene glycol and pentylene glycol, the glycerol content in the obtained product is 10% of the total mass, the hexylene glycol content is 2% of the total mass, and the pentylene glycol content is 2% of the total mass, and then sterilized at 100℃ for 30 min.

[0171] Comparative Example 2

[0172] The oat beta-glucan is prepared, and the specific steps are as follows:

[0173] A cosmetic grade 1% oat beta-glucan aqueous solution (purchased from Beijing Dongfangmiansen Biotechnology Co., Ltd.) is purchased on the market, which also contains 12% glycerol and 1% PEHG.

[0174] Comparative Example 3

[0175] Preparation of high-pressure homogenized oat beta-glucan, the specific steps are as follows:

[0176] The 1% by mass oat beta-glucan aqueous solution obtained in Comparative Example 2 was subjected to high-pressure homogenization, and 1 pass of high-pressure homogenization at 20 MPa gauge pressure was performed.

[0177] Comparative Example 4

[0178] Preparation of high-pressure homogenized oat beta-glucan, the specific steps are as follows:

[0179] The 1% by mass oat beta-glucan aqueous solution obtained in Comparative Example 2 was subjected to high-pressure homogenization, and 2 passes of high-pressure homogenization at 40 MPa gauge pressure were performed.

[0180] Comparative Example 5

[0181] Preparation of high-pressure homogenized oat beta-glucan, the specific steps are as follows:

[0182] The 1% by mass oat beta-glucan aqueous solution obtained in Comparative Example 2 was subjected to high-pressure shear homogenization, and homogenization was performed at 15000 r / min for 20 min.

[0183] The samples prepared in Examples 1-11 and Comparative Examples 1-5 were subjected to physicochemical index detection, and the results are shown in Table 1:

[0184] Physicochemical index detection method:

[0185] Color: The color was detected using a platinum-cobalt colorimeter (HI96727). First, ultra-pure water was used for zero adjustment, and then each example / comparative example product sample was added for color testing.

[0186] Turbidity: The turbidity was detected using an ISO portable turbidimeter (HI98713-01). Before use, a standard turbidity sample was used for calibration, and then each example / comparative example product sample was added for turbidity testing.

[0187] Table 1 Physicochemical indexes of samples of the present application and comparative examples

[0188] According to the requirements of the cosmetic industry, it is more desirable for the raw material to be clear and colorless or close to colorless, so that the appearance of the finished product is minimally affected in formulation application. Therefore, color less than 90 and turbidity less than 5 are preferred. Therefore, some of the comparative examples in the above groups are also optional in terms of appearance, but even if the appearance of different beta-glucan samples meets the standards or is almost the same, there may still be large differences in stability, efficacy, etc. in subsequent formulation applications.

[0189] The samples prepared in Examples 1-11 and Comparative Examples 1-5 were subjected to stability observation, and the results are shown in Table 2:

[0190] Physical and chemical index detection method:

[0191] The sample is placed at room temperature in the dark (25℃±2℃, 60%RH±10%RH) and observed every 30 days in a small high-definition photography studio (Chunying) using white light. If the sample shows precipitation or gel, it is recorded as unstable. Then the number of stable days is counted and recorded.

[0192] Table 2 Stability observation results of samples of the embodiments and comparative examples of the application

[0193] As can be seen from Table 2, the stability observation results of the embodiments of the application are significantly better than the stability results of the comparative examples. The stability observation results of the embodiments of the application refer to the actual observation days as of the filing date, and within the above time range, there is no instability phenomenon such as delamination, precipitation, turbidity, discoloration, etc. It does not mean that it can only maintain stability for the above time. Those skilled in the art should be able to infer from this that the flexible glucan product of the application can also maintain stability for a longer period of time. The stability observation results of Comparative Examples 1-5 refer to the fact that within the above time, delamination, precipitation, turbidity, discoloration and other instability phenomena have already occurred, which will affect the final efficacy expression and application. It can be seen that the flexible glucan product of the application has better stability, and further has a wider application prospect in terms of co-use with other raw materials in formula products, improvement of addition amount, etc.

[0194] Scanning electron microscope method:

[0195] Each embodiment and comparative example was sampled and freeze-dried (freeze dryer, Biomedica, FD-1A-80), then the freeze-dried sample was cut open, and the sample at the cross section was adhered to the sample holder using conductive tape. First, an ion sputtering instrument (Hitachi, E-1010) was used to perform a 60s gold spraying operation once under a vacuum of 5pa with a current value of 15mA. Each sample was repeated twice to increase the conductivity of the sample. Finally, a scanning electron microscope (Hitachi, S-3400N) was used to take pictures and record the morphology of the sample at 300X and 1000X under a voltage of 10KW.

[0196] The flexible glucan samples of the application were subjected to scanning electron microscope morphology characterization, and the results are shown in Figures 1-6:

[0197] As can be seen from Figures 1-6, the micro-morphology of the samples of Comparative Examples 1-4 has a clear lamellar structure, while the micro-morphology of the samples of embodiments 3 and 7 of the application can be seen to have a clear chain structure, and form a flexible space structure, with orderly extension between adjacent glucan chains, without stacking and accumulating into a lamellar structure.

[0198] Atomic force microscope method:

[0199] Each of the examples / Comparative Examples was sampled and freeze-dried, then configured into a 1 mg / mL solution, then diluted to 5 μg / mL, 10 μL of the diluted sample to be tested was dropped and deposited onto freshly cleaved mica, and dried in air at room temperature for 1.5 hours; the measurement parameters: before AFM imaging, a Petri dish was used for covering, using a magnetic alternating current (MAC) mode, using a Picoscan atomic force microscope for measurement.

[0200] The solution conformation characterization of the flexible glucan samples of the present application by atomic force microscopy was carried out, and the results are shown in Figures 7-8:

[0201] As can be seen from Figures 7-8, the micro three-dimensional structure of the sample of Comparative Example 2 is in scattered dots, and after stacking, a lamellar structure is formed, while the micro three-dimensional structure of the sample of Example 3 of the present application has a more abundant three-dimensional structure, in which the glucan molecules have obvious globular chain structures, including partial space network structures, which may be due to weak binding between flexible glucan chains under the action of some weak forces, such as intermolecular forces, hydrogen bonds, etc., which further promotes the excellent stability of the flexible glucan of the present application, and at the same time, such space network structure can better maintain the flexibility of the glucan chain, so that it can be easily used and better play its efficacy.

[0202] Cell scratch test method:

[0203] HaCaT cells were seeded into 6-well plates at a corresponding seeding density of 5*10 5 The cells were seeded into 6-well plates at a corresponding seeding density of 5*10 5 The cells were seeded into 6-well plates at a corresponding seeding density of 5*10

[0204] Scratch healing rate = (area before scratch - area after 24h scratch) / area before scratch * 100%

[0205] The flexible glucan samples obtained from Examples 1-11 and Comparative Examples 1-5 of the present application were subjected to efficacy characterization by cell scratch test, and the results are shown in Figures 9-10:

[0206] According to FIGS. 9-10, it can be seen that the cell repair of the samples of embodiments 1-11 of the present application is comprehensively improved compared with the samples of comparative examples 1-5, and the cell repair of some of the embodiment samples has reached the level of EGF (epidermal growth factor). EGF has multiple biological activities, can cause a series of biochemical activities in cells, can stimulate epidermis and epithelial cells, promote epidermal proliferation and keratinization, can strongly promote cell division, enhance cell activity, promote metabolism, and promote the synthesis of hyaluronic acid and glycoprotein. EGF can also promote the healing of skin and mucosal wounds, has the effects of preventing and treating ulcers and anti-inflammatory analgesia, is widely used in the treatment of burns, scalds, surgical injuries, mechanical injuries, skin ulcers, laser beauty, etc., and can effectively inhibit the growth of acne and acne, and protect the skin and mucosa from or less mechanical and chemical damage. EGF has a wide application in the field of wound care (such as burns, scalds, abrasions, etc.) due to its excellent performance, but the injection of EGF in the medical beauty industry has a huge safety risk, which can cause uncontrollable growth of local skin tissue and further cause the formation of nodules and lumps, and in severe cases can cause disfigurement. In view of this, the State Drug Administration of China has listed EGF as a prohibited ingredient. The flexible dextran product provided in the present application can be said to be a safe, mild, and plant-derived effective substitute for EGF.

[0207] Cell anti-inflammatory experiment method:

[0208] RAW264.7 cells were inoculated in a 24-well plate at a corresponding cell density of 3*10 5 cells / well, and incubated in a CO2 cell incubator overnight. The next day, the sample of the embodiment / comparative example product to be tested was sampled and dissolved in high-sugar DMEM medium (gibio, C11965500BT), and configured to use 0.25% (mass fraction) of different embodiment and comparative example product sample solutions to treat cells and incubate for 24 h. On the third day, 10 ng / mL of LPS was added for inflammation induction and incubation for 24 h. On the fourth day, the supernatant of each group was detected for inflammatory factors according to the operation of the TNF-α ELISA kit.

[0209] The flexible dextran samples obtained from embodiments 1-11 and comparative examples 1-5 of the present application were subjected to cell anti-inflammatory efficacy characterization, and the results are shown in FIG. 11.

[0210] According to FIG. 11, it can be seen that the anti-inflammatory effect of the samples of embodiments 1-11 of the present application is comprehensively improved compared with the samples of comparative examples 1-5, and even reaches / exceeds the level of dexamethasone, an anti-inflammatory drug, at a general dosage (mass fraction of 0.001%, higher dosage will have cytotoxicity). The flexible dextran product of the present application has a significant advantage in anti-inflammatory effect.

[0211] Each example and comparative example was sampled and freeze-dried, the freeze-dried powder was subjected to monosaccharide composition detection, and the remaining sample was configured into a 1 mg / mL solution for flexible structure characterization, with the specific methods as follows:

[0212] Monosaccharide composition detection: a clean chromatographic bottle was weighed, an appropriate amount of sample was added, 1 ml of 2M TFA acid solution was added, and heated at 121°C for 2 hours. Nitrogen was blown and dried. 99.99% methanol was added for cleaning, and then dried, and the methanol cleaning was repeated 2-3 times. Sterile water was added for dissolution, and transferred into a chromatographic bottle for testing.

[0213] The chromatographic system used was a Thermo ICS 5000+ ion chromatography system (ICS 5000+, Thermo Fisher Scientific, USA), which used an electrochemical detector to analyze and detect the monosaccharide components.

[0214] A DionexTM CarboPacTM PA20 (150*3.0mm, 10μm) liquid chromatography column was used; the sample size was 5μL. The mobile phase A (H2O), the mobile phase B (0.1M NaOH), the mobile phase C (0.1M NaOH, 0.2M NaAc), the flow rate was 0.5mL / min; the column temperature was 30°C; the elution gradient was: 0min A phase / B phase / C phase (95:5:0, V / V), 26min A phase / B phase / C phase (85:5:10, V / V), 42min A phase / B phase / C phase (85:5:10, V / V), 42.1min A phase / B phase / C phase (60:0:40, V / V), 52min A phase / B phase / C phase (60:40:0, V / V), 52.1min A phase / B phase / C phase (95:5:0, V / V), 60min A phase / B phase / C phase (95:5:0, V / V).

[0215] The monosaccharide composition results of each group of samples were obtained by comparing with the standard spectrum.

[0216] Flexible structure characterization:

[0217] The prepared 1 mg / mL sample solution of each example / contrast example was filtered through a 0.45 μm filter membrane for testing. The chromatographic system was a gel chromatography-differential-multiple angle laser light scattering system, the liquid phase system was U3000 (Thermo, USA), the differential detector was Optilab T-rEX (Wyatt technology, CA, USA), and the laser light scattering detector was DAWN HELEOS II (Wyatt technology, CA, USA). The chromatographic conditions were as follows: an Ohpak SB-805HQ (300*8 mm) and an Ohpak SB-803HQ (300*8 mm) were connected in series as the gel exclusion chromatography column. The column temperature was 45 °C, the injection volume was 100 μL, the mobile phase A was 0.02% (mass fraction) NaN3 and 0.1 M NaNO3, the flow rate was 0.6 mL / min, and the elution gradient was isocratic for 75 min. The chromatographic data were processed by using software ASTRA 6.1 to obtain a molecular configuration diagram with the molar mass (g / mol) as the abscissa and the root mean square radius (R.M.S.Radius, nm) as the ordinate, and the slope was used as the molecular conformation parameter. When the slope was less than 0.5, the molecules were mostly spherical; when the slope was greater than 0.8, the molecules were mostly rigid rod-like structures; and when 0.8≥ the slope ≥0.5, the molecules were mostly flexible random coil configurations.

[0218] The flexible dextran prepared from the examples and contrast examples of the application was subjected to monosaccharide composition and flexible structure characterization, and the results are shown in Tables 3 and 4.

[0219] Table 3: Monosaccharide composition detection results of the examples and contrast examples of the application

[0220] As can be seen from Table 3, the monosaccharide composition results of the contrast examples and the examples of the application are similar, which proves that the modification method of the application does not cause changes in the monosaccharide composition of the sample and has little effect on the primary structure of polysaccharide.

[0221] Table 4: Molecular conformation detection results of the examples and contrast examples of the application

[0222] As can be seen from Table 4, the molecular configuration parameter of the contrast examples is 0.27<0.5, indicating that it is a spherical configuration, while the molecular configuration parameter of the examples of the application is 0.62, which is between 0.5 and 0.8, indicating that it is a flexible random coil configuration. The molecular weight of the contrast examples and the examples has little difference, which proves that the application effectively regulates the structure of dextran molecules to convert them from the original spherical configuration to the flexible random coil configuration, thereby obtaining flexible dextran.

[0223] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present application, and are not intended to limit the present application; although the present application has been described in detail with reference to the above embodiments, those skilled in the art should understand that the technical solutions recorded in the above embodiments can be modified, or some or all of the technical features can be replaced by equivalents; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the scope of the technical solutions of the embodiments of the present application.

Claims

1. A product containing a flexible glucan, characterized in that, The flexible glucan molecules in the product containing flexible glucan have a globular chain structure, and the molecular conformation parameter is 0.5-0.

8.

2. A product containing flexible glucan according to claim 1, characterized in that, The glucan comprises β-glucan; Preferably, the glucan comprises cereal β-glucan; Preferably, the glucan comprises one or more of oat β-glucan, barley β-glucan and barley β-glucan.

3. A method for the preparation of a product containing a flexible glucan, characterized in that, The treatment of the material containing glucan by high-pressure microfluidization is performed.

4. A method of preparing a product containing flexible glucan according to claim 3, characterized in that, The glucan comprises β-glucan; Preferably, the glucan comprises cereal β-glucan; Preferably, the glucan comprises one or more of oat β-glucan, barley β-glucan and barley β-glucan; Preferably, the material containing glucan comprises an extract containing glucan; Preferably, in the treatment of the material containing glucan by high-pressure microfluidization, the mass ratio of the material containing glucan to the fluid is 1:(12-70), and further preferably 1:14; Preferably, the fluid comprises water; Preferably, the high-pressure microfluidization pressure is 80-140 MPa in gauge pressure, and further preferably 120 MPa in gauge pressure; Preferably, the high-pressure microfluidization is performed 1-6 times, and further preferably 2 times. Preferably, the high-pressure microfluidization feed temperature is 0-45℃; further preferably 25℃.

5. A method of preparing a product containing flexible glucan according to claim 3, characterized in that, The material containing glucan is heated and dissolved before high-pressure microfluidization treatment; Preferably, the heating and dissolving temperature is 70-90℃; further preferably, the heating and dissolving temperature is 80-85℃.

6. A method of preparing a product containing flexible glucan according to claim 3, characterized in that, The preparation method further comprises at least one of decolorization, desalination and protein removal.

7. A method of preparing a product containing flexible glucan according to claim 6, characterized in that, The number of decolorization is 1-3 times, and preferably 2 times; Preferably, the total amount of decolorizing agent added in the decolorization is 2%-5% of the total mass of the material to be decolorized, and further preferably 2-3%; Preferably, the decolorizing agent comprises activated carbon; Preferably, the decolorization time of each decolorization is 30-60 min; Preferably, the decolorization temperature of each decolorization is 75-85℃; Further preferably, the number of decolorization is 2 times, 3% of the total mass of the material to be decolorized is added as decolorizing agent in the first decolorization, and the decolorization is performed at 80-85℃ for 30 min, and 2% of the total mass of the material to be decolorized is added as decolorizing agent activated carbon in the second decolorization, and the decolorization is performed at 80-85℃ for 30 min; Preferably, cation and anion exchange resins are used for desalination; Preferably, the material flow rate in the desalination process is 1-3 t / h, and further preferably, the material flow rate in the desalination process is 2-3 t / h.

8. A method of preparing a product containing flexible glucan according to claim 5 or 6, characterised in that, The obtained product is filtered and compounded to obtain a product containing flexible glucan; Preferably, one or more of hexylene glycol, pentylene glycol, glycerol and PEHG is used as a compounding solvent for compounding; Preferably, sterilization is performed after compounding.

9. A composition comprising a flexible glucan, characterized in that, A product containing flexible glucan according to any one of claims 1-2, and / or a product containing flexible glucan prepared by the preparation method according to any one of claims 3-8.

10. Use of a product containing flexible glucan as defined in any one of claims 1 to 2, a product containing flexible glucan prepared by a process as defined in any one of claims 3 to 8 or a composition containing flexible glucan as defined in claim 9 for the preparation of a pharmaceutical product and / or a cosmetic product.

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