Collagen-containing composition as well as preparation method and application thereof

Abstract

The invention relates to a collagen-containing composition and a preparation method and application thereof, the composition comprises collagen and a plant extract, and the plant extract is prepared from sambucus chinensis, perigord truffle, citrus peel, lycium ruthenicum and rhodomyrtus tomentosa. The prepared composition is powder and does not contain preservatives, the addition amount is easy to control, and the composition is easy to store and transport, the plant extract can improve the repairing effect of the composition while improving the stability, and the sambucus chinensis, the perigord truffle, the citrus peel, the lycium ruthenicum and the rhodomyrtus tomentosa have a certain synergistic effect on the effects.

Description

Technical Field

[0001] The invention belongs to the technical field of cosmetics and relates to a composition containing collagen and a preparation method and application thereof. Background Art

[0002] Collagen is an extremely important structural protein in the human body. It is widely present in tissues such as skin, bones, tendons, and ligaments. It plays a key role in maintaining the structural integrity, elasticity, and normal physiological functions of these tissues. In aqueous dosage forms containing collagen, collagen is usually dissolved or dispersed in the aqueous phase system. However, aqueous dosage forms have many limitations. From a stability perspective, the stability of collagen in aqueous solutions is affected by many factors, such as pH, temperature, and interactions with other ingredients. It is prone to degradation, resulting in a reduction in the content of active ingredients. In order to ensure the solubility of collagen in aqueous solutions, it is often necessary to add a certain amount of additives such as solubilizers. These additives may have potential adverse effects on the human body, and may also interfere with the interaction between collagen and skin or human tissues, weakening its original physiological effects.

[0003] The powder dosage form containing collagen is made into powder by a specific drying process, which can exist alone or be mixed with other suitable excipients. The stability of the powder is relatively high and high concentration of collagen can be added. Based on the advantages of the powder, it is expected to become a better dosage form choice. Therefore, it is urgent to provide a collagen powder composition. Summary of the invention

[0004] In view of the deficiencies of the prior art, the object of the present invention is to provide a composition containing collagen and a preparation method and application thereof.

[0005] In order to achieve the purpose of the invention, the present invention adopts the following technical solutions:

[0006] In a first aspect, the present invention provides a composition containing collagen, wherein the composition comprises collagen and a plant extract, wherein raw materials for preparing the plant extract comprise elderberry, black truffle, citrus peel, black wolfberry and myrtle.

[0007] The composition prepared by the present invention is a powder, does not contain preservatives, the addition amount is easy to control, and it is easy to store and transport. The plant extract can improve the stability and the repair effect of the composition at the same time. Elderberry, black truffle, citrus peel, black wolfberry and myrtle have a certain synergistic effect on the above effects.

[0008] Preferably, the composition comprises 1-3 parts of collagen and 2-4 parts of plant extract by mass.

[0009] The mass proportions of collagen can be selected as 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts etc., and the mass proportions of plant extract can be selected as 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts, 3.2 parts, 3.4 parts, 3.6 parts, 3.8 parts, 4 parts etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here one by one.

[0010] Preferably, the raw materials for preparing the plant extract include, by mass, 2-4 parts of elderberry, 1-2 parts of black truffle, 1-2 parts of citrus peel, 2-4 parts of black wolfberry, and 1-3 parts of myrtle.

[0011] The mass proportion of comfrey can be 2 parts, 2.2 parts, 2.5 parts, 2.8 parts, 3 parts, 3.2 parts, 3.4 parts, 3.6 parts, 3.8 parts, 4 parts, etc. The mass proportion of black truffle can be 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts, 1.7 parts, 1.8 parts, 1.9 parts, 2 parts, etc. The mass proportion of citrus peel can be 1.1 parts, 1.2 parts, 1.3 parts, 1.4 parts, 1.5 parts, 1.6 parts, The mass portions of black wolfberry can be selected as 2 parts, 2.2 parts, 2.5 parts, 2.8 parts, 3 parts, 3.2 parts, 3.4 parts, 3.6 parts, 3.8 parts, 4 parts, etc. The mass portions of myrtle can be 1 part, 1.2 parts, 1.4 parts, 1.6 parts, 1.8 parts, 2 parts, 2.2 parts, 2.4 parts, 2.6 parts, 2.8 parts, 3 parts, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here one by one.

[0012] Preferably, the plant extract is prepared by a method comprising the following steps: mixing the preparation raw material with water, extracting, centrifuging, and filtering.

[0013] Preferably, the mass ratio of the preparation raw materials to water is 4:(10-15), wherein the specific point values ​​in (10-15) can be selected as 10, 10.5, 11, 11.5, 12, 12.5, 13, 13.5, 14, 14.5, 15, etc. Other specific point values ​​within the above numerical range can be selected, and they will not be described one by one here.

[0014] Preferably, the leaching temperature is 65-75°C and the leaching time is 0.5-1.5h.

[0015] The temperature can be selected as 65℃, 66℃, 67℃, 68℃, 69℃, 70℃, 71℃, 72℃, 73℃, 74℃, 75℃, etc. The time can be selected as 0.5h, 0.8h, 1h, 1.2h, 1.5h, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0016] Preferably, the pore size of the ceramic membrane used for filtration is 100-150nm, for example 100nm, 105nm, 110nm, 115nm, 120nm, 125nm, 130nm, 135nm, 140nm, 145nm, 150nm, etc. Other specific point values ​​within the above numerical range can be selected, which will not be described here one by one.

[0017] Preferably, the collagen is prepared by a method comprising the following steps:

[0018] (1) activating Pichia pastoris and inoculating the activated Pichia pastoris into a fermentation medium for fermentation to obtain a fermentation product;

[0019] (2) Centrifuge the fermentation product, take the supernatant and filter it.

[0020] Preferably, the fermentation medium comprises, by mass percentage, 1-5% glucose, 0.5-2% tryptone, 0.1-1% yeast extract powder, 0.01-0.1% magnesium sulfate, 0.05-0.2% potassium dihydrogen phosphate and 93-98.5% water.

[0021] The mass percentage of glucose can be selected from 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, etc. The mass percentage of tryptone can be selected from 0.5%, 0.8%, 1%, 1.2%, 1.5%, 1.8%, 2%, etc. The mass percentage of yeast extract can be selected from 0.1%, 0.3%, 0.5%, 0.7%, 1%, etc. The mass percentage of magnesium sulfate can be selected from 0.01 The mass percentage of potassium dihydrogen phosphate can be selected as 0.05%, 0.08%, 0.1%, 0.12%, 0.15%, 0.18%, 0.2% etc., the mass percentage of water can be selected as 93%, 94%, 95%, 96%, 97%, 98%, 98.5% etc., and other specific point values ​​within the above numerical range can be selected, so they will not be described here one by one.

[0022] Preferably, the fermentation temperature is 25-30°C, the fermentation time is 45-55h, and the rotation speed is 120-180rpm.

[0023] The fermentation temperature can be selected as 25℃, 26℃, 27℃, 28℃, 29℃, 30℃, etc. The time can be selected as 45h, 46h, 47h, 48h, 49h, 50h, 51h, 52h, 53h, 54h, 55h, etc. The speed can be selected as 120rpm, 130rpm, 140rpm, 150rpm, 160rpm, 170rpm, 180rpm, etc. Other specific point values ​​within the above numerical range can be selected, so they will not be elaborated here.

[0024] Preferably, the pore size of the ceramic membrane used for filtration is 80-120nm, for example 80nm, 85nm, 90nm, 95nm, 100nm, 105nm, 110nm, 115nm, 120nm, etc. Other specific point values ​​within the above numerical range can be selected and will not be described here one by one.

[0025] Preferably, the dosage form of the composition comprises a powder.

[0026] In a second aspect, the present invention provides a method for preparing the composition containing collagen according to the first aspect, the preparation method comprising: mixing the plant extract with the collagen, and drying.

[0027] Preferably, the drying method is freeze drying.

[0028] In a third aspect, the present invention provides a use of the composition according to the first aspect in preparing a product with repair efficacy.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] The composition prepared by the present invention is a powder, does not contain preservatives, the addition amount is easy to control, and it is easy to store and transport. The plant extract can improve the stability and the repair effect of the composition at the same time. Elderberry, black truffle, citrus peel, black wolfberry and myrtle have a certain synergistic effect on the above effects. DETAILED DESCRIPTION

[0031] In order to further explain the technical means and effects adopted by the present invention, the technical solution of the present invention is further described below in conjunction with the preferred embodiments of the present invention, but the present invention is not limited to the scope of the embodiments.

[0032] The sources of the functional ingredients contained in the products involved in the following embodiments and comparative examples are as follows (only the functional ingredients are reflected, and the necessary auxiliary ingredients contained in other commercially available raw materials are not repeated):

[0033] Fermentation medium: in terms of mass percentage, it includes 2% glucose, 1% tryptone, 0.5% yeast extract powder, 0.05% magnesium sulfate, 0.1% potassium dihydrogen phosphate and the balance water, with a pH of about 6, and sterilized at 121° C. for 20 min.

[0034] Preparation Example 1

[0035] This preparation example provides a plant extract, which includes 3 parts of sagebrush, 1.5 parts of black truffle, 1.5 parts of citrus peel, 3 parts of black wolfberry, 2 parts of myrtle and 27.5 parts of water in parts by mass.

[0036] The preparation method thereof comprises:

[0037] Mix elderberry, black truffle, citrus peel, black wolfberry, myrtle and water, extract at 70°C for 1 hour, and filter through a 120nm ceramic membrane to obtain the product.

[0038] Preparation Example 2

[0039] This preparation example provides a plant extract, which includes, by weight, 2 parts of elderberry, 1 part of black truffle, 2 parts of citrus peel, 4 parts of black wolfberry, 1 part of myrtle and 37.5 parts of water.

[0040] The preparation method thereof comprises:

[0041] Mix elderberry, black truffle, citrus peel, black wolfberry, myrtle and water, extract at 65° C. for 1.5 h, and filter using a 100 nm ceramic membrane to obtain the product.

[0042] Preparation Example 3

[0043] This preparation example provides a plant extract, which includes 4 parts of sage, 2 parts of black truffle, 1 part of citrus peel, 2 parts of black wolfberry, 3 parts of myrtle and 45 parts of water in parts by mass.

[0044] The preparation method thereof comprises:

[0045] Mix elderberry, black truffle, citrus peel, black wolfberry, myrtle and water, extract at 75°C for 0.5h, and filter through a 150nm ceramic membrane to obtain the product.

[0046] Preparation Example 4

[0047] This preparation example provides a collagen protein, which is prepared by a method comprising the following steps:

[0048] (1) The Pichia pastoris GS115 strain was transferred to a slant medium for activation and cultured at 28°C until the mycelium covered the slant; the slant strain was then transferred to a liquid medium and cultured for 2 days to obtain a fermentation seed liquid;

[0049] (2) The Pichia pastoris seed solution was inoculated into the fermentation medium at an inoculum rate of 10%, and the fermentation was carried out at 28° C. and 160 rpm for 50 h. The fermentation product was centrifuged to remove the bacterial cells, and the fermentation product was obtained after passing through a 100 nm ceramic membrane and stored at 4° C.

[0050] Preparation Example 5

[0051] This preparation example provides a collagen protein, which is prepared by a method comprising the following steps:

[0052] (1) The Pichia pastoris GS115 strain was transferred to a slant medium for activation and cultured at 28°C until the mycelium covered the slant; the slant strain was then transferred to a liquid medium and cultured for 2 days to obtain a fermentation seed liquid;

[0053] (2) The Pichia pastoris seed solution was inoculated into the fermentation medium at an inoculum rate of 10%, and the fermentation was carried out at 25° C. and 180 rpm for 55 h. The fermentation product was centrifuged to remove the bacterial cells, and the fermentation product was obtained after passing through a 120 nm ceramic membrane and stored at 4° C.

[0054] Preparation Example 6

[0055] This preparation example provides a collagen protein, which is prepared by a method comprising the following steps:

[0056] (1) The Pichia pastoris GS115 strain was transferred to a slant medium for activation and cultured at 28°C until the mycelium covered the slant; the slant strain was then transferred to a liquid medium and cultured for 2 days to obtain a fermentation seed liquid;

[0057] (2) The Pichia pastoris seed solution was inoculated into the fermentation medium at an inoculum rate of 10%, and the fermentation was carried out at 30° C. and 120 rpm for 45 h. The fermentation product was centrifuged to remove the bacterial cells, and the fermentation product was obtained after passing through an 80 nm ceramic membrane and stored at 4° C.

[0058] Preparation Example 7

[0059] This preparation example provides a plant extract, which differs from Preparation Example 1 only in that the preparation method is different.

[0060] The preparation method is as follows: mix elderberry, black truffle, citrus peel, black wolfberry, myrtle and water, extract at 70°C for 1 hour, filter with a 120nm ceramic membrane, and dry to obtain the product.

[0061] Preparation Example 8

[0062] This preparation example provides a collagen, which differs from Preparation Example 1 only in the preparation method.

[0063] The step (2) of the preparation method is "inoculating Pichia pastoris seed liquid into a fermentation medium at a 10% inoculation rate, fermenting at 28° C. and 160 rpm for 50 hours, centrifuging the fermentation product to remove the bacteria, passing the fermentation product through a 100 nm ceramic membrane, and drying to obtain the product", and the other operations remain unchanged.

[0064] Comparative Preparation Example 1

[0065] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain sagebrush, and its reduced mass is proportionally distributed to the mass of black truffle, citrus peel, black wolfberry, and myrtle, while other ingredients and contents remain unchanged.

[0066] The preparation method is similar to that of Preparation Example 1.

[0067] Comparative Preparation Example 2

[0068] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain black truffle, and its reduced mass is proportionally distributed to the mass of elderberry, citrus peel, black wolfberry, and myrtle, while other ingredients and contents remain unchanged.

[0069] The preparation method is similar to that of Preparation Example 1.

[0070] Comparative Preparation Example 3

[0071] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain citrus peel, and its reduced mass is proportionally distributed to the mass of elderberry, black truffle, black wolfberry, and myrtle, while other ingredients and contents remain unchanged.

[0072] The preparation method is similar to that of Preparation Example 1.

[0073] Comparative Preparation Example 4

[0074] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain black wolfberry, and its reduced mass is proportionally distributed to the mass of elderberry, black truffle, citrus peel, and myrtle, while other ingredients and contents remain unchanged.

[0075] The preparation method is similar to that of Preparation Example 1.

[0076] Comparative Preparation Example 5

[0077] This comparative preparation example provides a plant extract, which differs from Preparation Example 1 only in that it does not contain myrtle, and its reduced mass is proportionally distributed to the mass of elderberry, black truffle, citrus peel, and black wolfberry, while other ingredients and contents remain unchanged.

[0078] The preparation method is similar to that of Preparation Example 1.

[0079] Example 1

[0080] This embodiment provides a composition containing collagen, which includes 3 parts of plant extract prepared in Preparation Example 1 and 2 parts of collagen prepared in Preparation Example 4 in terms of mass fractions, and the mass fractions are all mass fractions of solutes in the solution.

[0081] The preparation method thereof comprises:

[0082] The plant extract is mixed with collagen and freeze-dried.

[0083] Example 2

[0084] This embodiment provides a composition containing collagen, which includes 2 parts of the plant extract prepared in Preparation Example 2 and 3 parts of the collagen prepared in Preparation Example 5 in terms of mass fractions, and the mass fractions are all mass fractions of the solute in the solution.

[0085] The preparation method is similar to that of Example 1.

[0086] Example 3

[0087] This embodiment provides a composition containing collagen, which includes 4 parts of the plant extract prepared in Preparation Example 3 and 1 part of the collagen prepared in Preparation Example 6 in terms of mass fractions, and the mass fractions are all mass fractions of the solute in the solution.

[0088] The preparation method is similar to that of Example 1.

[0089] Example 4

[0090] This embodiment provides a composition containing collagen, which is different from Embodiment 1 only in the preparation method.

[0091] The preparation method comprises the following steps: mixing the plant extract with collagen to obtain the collagen.

[0092] Comparative Examples 1-5

[0093] This comparative example provides 5 compositions containing collagen, which differ from Example 1 only in that the plant extract prepared in Preparation Example 1 is replaced by the plant extracts prepared in Comparative Preparation Examples 1-5 in equal amounts, and other components and contents remain unchanged.

[0094] The preparation method is similar to that of Example 1.

[0095] Test Example 1

[0096] Stability test

[0097] The obtained products were placed at a temperature of 25±2°C and a relative humidity of 60±5% for different periods of time, and the appearance of the products was observed with the naked eye. The results are shown in Table 1.

[0098] Table 1

[0099]

[0100]

[0101] It can be seen from the data in Table 1 that the stability of the powder dosage form is better than that of the liquid dosage form, and the addition of plant extracts improves the stability of the product. Elderberry, black truffle, citrus peel, black wolfberry and myrtle have a certain synergistic effect in improving product stability.

[0102] Test Example 2

[0103] Cytotoxicity:

[0104] Samples to be tested: samples prepared in Examples 1-4, Comparative Examples 1-5, and Preparation Examples 1 and 4 were placed at 25° C. and a relative humidity of 65% for 3 months. The liquid samples after being placed for 3 months were freeze-dried to obtain solid powders.

[0105] BJ cells were placed in DMEM high-glucose medium containing 10% fetal bovine serum and 1% double antibody, and cultured at 37°C and 5% CO2. When the cells were in good growth condition, 1×10 5 Each concentration was repeated 5 times, and BJ cells were inoculated in a 96-well plate, with 150 μL of culture medium added to each well, and incubated at 37°C for 24 hours. Except for the normal group, the culture medium containing 1% of the sample to be tested was replaced and cultured for 24 hours. Then, 15 μL of CCK-8 reagent was added to each well, and the cells were incubated at 37°C in the dark for 0.5 hours. The OD value was detected at 450 nm, and the cell viability was calculated.

[0106] Among them: the OD of the normal group is the absorbance of the cells when no sample to be tested is added to the culture medium, and the OD of the blank group is the absorbance of the 96-well plate itself.

[0107]

[0108] Table 2

[0109] Survival rate (%) Preparation Example 1 92.30 Preparation Example 4 91.81 Example 1 96.53 Example 2 95.32 Example 3 94.97 Example 4 94.65 Comparative Example 1 93.17 Comparative Example 2 93.64 Comparative Example 3 92.69 Comparative Example 4 93.72 Comparative Example 5 92.48

[0110] It can be seen from the data in Table 2 that the composition of the present application has low cytotoxicity and high safety.

[0111] Test Example 3

[0112] Cell Repair

[0113] Samples to be tested: samples prepared in Examples 1-4, Comparative Examples 1-5, and Preparation Examples 1, 4, 7, and 8, which were placed at 25° C. and a relative humidity of 65% for 3 months; the liquid samples after being placed for 3 months were freeze-dried to obtain solid powders.

[0114] BJ cells were placed in DMEM high-glucose medium containing 10% fetal bovine serum and 1% double antibody, and cultured at 37°C and 5% CO2. When the cells were growing well, 5×10 cells were added to each well. 5 Each well was inoculated with 2 mL of culture medium and incubated at 37°C for 24 h. After making a cross scratch, the scratch distance was measured. Except for the blank group, the culture medium containing 1% of the sample to be tested was replaced and the scratch distance was measured again after culturing for 24 h.

[0115]

[0116] Table 3

[0117] Repair rate (%) Preparation Example 1 10.37 Preparation Example 4 15.48 Preparation Example 7 13.89 Preparation Example 8 16.92 Example 1 21.36 Example 2 18.64 Example 3 18.25 Example 4 17.54 Comparative Example 1 16.57 Comparative Example 2 16.32 Comparative Example 3 15.87 Comparative Example 4 16.01 Comparative Example 5 15.94 Blank Group 5.94 10% culture medium 9.34

[0118] It can be seen from the experimental results in Table 3 that the present application has a good cell repair effect. After being placed for a period of time, the efficacy of the composition in the form of a powder is better than that of the aqueous solution, indicating that the powder is more stable than the aqueous solution. The addition of the plant composition can not only improve the stability, but also improve the repair efficacy of the composition, and there is a certain synergistic effect between the several components.

[0119] The applicant declares that the present invention illustrates a composition containing collagen and its preparation method and application through the above-mentioned embodiments, but the present invention is not limited to the above-mentioned embodiments, that is, it does not mean that the present invention must rely on the above-mentioned embodiments to be implemented. Those skilled in the art should understand that any improvement of the present invention, equivalent replacement of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific methods, etc., all fall within the protection scope and disclosure scope of the present invention.

[0120] The preferred embodiments of the present invention are described in detail above. However, the present invention is not limited to the specific details in the above embodiments. Within the technical concept of the present invention, a variety of simple modifications can be made to the technical solution of the present invention, and these simple modifications all belong to the protection scope of the present invention.

[0121] It should also be noted that the various specific technical features described in the above specific embodiments can be combined in any suitable manner without contradiction. In order to avoid unnecessary repetition, the present invention will not further describe various possible combinations.

Claims

1. A composition containing collagen, characterized in that: The composition comprises collagen and plant extracts, and raw materials for preparing the plant extracts comprise elderberry, black truffle, citrus peel, black wolfberry and myrtle.

2. The composition containing collagen according to claim 1, characterized in that The composition comprises 1-3 parts of collagen and 2-4 parts of plant extract by mass; Preferably, the raw materials for preparing the plant extract include, by mass, 2-4 parts of elderberry, 1-2 parts of black truffle, 1-2 parts of citrus peel, 2-4 parts of black wolfberry, and 1-3 parts of myrtle.

3. The collagen-containing composition according to claim 1 or 2, characterized in that: The plant extract is prepared by a method comprising the following steps: mixing the raw material with water for extraction, centrifuging, and filtering to obtain the plant extract.

4. The collagen-containing composition according to claim 3, characterized in that The mass ratio of the preparation raw materials to water is 4:(10-15); Preferably, the leaching temperature is 65-75°C and the time is 0.5-1.5h; Preferably, the pore size of the ceramic membrane used for filtration is 100-150 nm.

5. The composition containing collagen according to any one of claims 1 to 4, characterized in that: The collagen is prepared by a method comprising the following steps: (1) activating Pichia pastoris and inoculating the activated Pichia pastoris into a fermentation medium for fermentation to obtain a fermentation product; (2) Centrifuge the fermentation product, take the supernatant and filter it.

6. The collagen-containing composition according to claim 5, characterized in that The fermentation medium comprises, by mass percentage, 1-5% glucose, 0.5-2% tryptone, 0.1-1% yeast extract powder, 0.01-0.1% magnesium sulfate, 0.05-0.2% potassium dihydrogen phosphate and 93-98.5% water; Preferably, the fermentation temperature is 25-30°C, the fermentation time is 45-55h, and the rotation speed is 120-180rpm; Preferably, the pore size of the ceramic membrane used for filtration is 80-120 nm.

7. The composition containing collagen according to any one of claims 1 to 6, characterized in that: The dosage form of the composition includes powder.

8. The method for preparing a composition containing collagen according to any one of claims 1 to 7, characterized in that: The method comprises: mixing the plant extract with collagen and drying the mixture.

9. The preparation method according to claim 8, characterized in that: The drying method is freeze drying.

10. Use of the composition according to any one of claims 1 to 7 in preparing a product with repair efficacy.

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