Water-in-oil nano-emulsion loaded with collagen hydrolyzed peptide and preparation method of water-in-oil nano-emulsion

Abstract

The invention discloses a water-in-oil nano-emulsion loaded with collagen hydrolyzed peptide and a preparation method of the water-in-oil nano-emulsion. The nano-emulsion is prepared from the following components in percentage by mass: 28 to 36 percent of PEG-40 hydrogenated castor oil, 18 to 2526 percent of polyglycerol ricinoleate, 6 to 8 percent of water-soluble glue protein hydrolyzed peptide and the balance of GTCC (Glycerol Tricarbonyl Cellulose) which is supplemented until the mass of the water-in-oil nano-emulsion is 100 percent. The method reduces the cost problem of an emulsification method, provides a basis for wide application of the method in different fields, and has good application prospects and industrial values.

Description

Technical Field

[0001] The invention relates to the field of cosmetics and cosmetic preparation, and in particular to a method for preparing a water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides. Background Art

[0002] Collagen hydrolyzed peptides, also known as collagen peptides, are a peptide mixture between protein and amino acids formed by the disintegration and breakage of collagen molecular chains through hydrolysis. The moisturizing, repairing and anti-aging effects of small molecule collagen hydrolyzed peptides are well known. It has become one of the popular raw materials in the cosmetics industry and is a new trend in cosmetics raw materials.

[0003] Oil nourishing skin is a skin care concept, which refers to the use of oily substances to moisturize and maintain the skin. This method has received more and more attention in recent years, because oil can form a protective film on the surface of the skin, help lock in moisture, and provide deep moisturizing. Although water-soluble skin care products can quickly replenish moisture on the surface of the skin, their moisturizing effect is often not as long-lasting as oil-soluble products; at the same time, oil products can promote the absorption of active ingredients in other skin care products, making the skin care effect better. However, most collagen hydrolyzed peptides are water-soluble and will not dissolve in most oils or non-polar solvents. At present, there are several main ways to solve the oil solubility problem of collagen hydrolyzed peptides, including chemical modification, use of amphiphilic carriers, nano-microemulsions and emulsification technology. Chemical modification can precisely control the surface properties of peptides through chemical modification. The modified peptides may have better bioavailability or functionality, but the process is more complicated, the biological activity needs to be verified multiple times and the cost is high. The use of amphiphilic carriers such as liposomes and microcapsules can carry hydrophilic and lipophilic substances at the same time, but their preparation is difficult and the stability of the carrier is also a problem. Although most nano-microemulsions can provide higher dispersibility and better absorption efficiency, they all require high temperature, high pressure or high shear force, which affects the stability and efficacy of active ingredients. In addition, the oil solubility of water-soluble active ingredients is still a big challenge.

[0004] Water-in-oil (W / O) nanoemulsion is a common method to solve the oil solubility problem of active ingredients. The inner phase of W / O nanoemulsion is water phase and the outer phase is oil phase. The interfacial tension between the water phase and the oil phase is reduced by the emulsifier, and the additional shear force is applied by the homogenizer. The water phase is slowly emulsified into the oil phase. After the emulsification is completed, the originally incompatible water phase and oil phase finally form a uniform emulsion of water-in-oil dosage form through emulsification. In the pharmaceutical industry, it has significant advantages as an advanced drug delivery system. In the cosmetics industry, W / O nanoemulsion can be used to deliver oil-soluble antioxidants, vitamin E and other ingredients to improve their skin penetration ability. Due to the high cost of natural emulsifiers or composite emulsifiers, most of the commonly used emulsifiers on the market are single emulsifiers or synthetic emulsifiers, such as sorbitan laurate, polysorbates, sorbitan oleate, sodium dodecyl sulfate, etc., which may cause problems such as system instability, poor tolerance and insufficient sensory characteristics, and also have disadvantages such as safety, biocompatibility or environmental unfriendliness. In addition, the addition of emulsifiers usually involves high-energy emulsification processes such as high temperature or high shear force, which can easily lead to high collagen production costs and damage its activity.

[0005] Therefore, there is an urgent need in the art for a method for preparing an oil-in-water nanoemulsion with safe emulsifier types, low production cost and low energy emulsification to solve the oil-solubility problem of collagen hydrolyzed peptides. Summary of the invention

[0006] In order to overcome the shortcomings of the prior art, the present application provides a method for preparing an oil-in-water nanoemulsion loaded with collagen hydrolyzed peptides, which has good biocompatibility and low review cost, and can give full play to the moisturizing, repairing, anti-aging and other effects of collagen hydrolyzed peptides.

[0007] The invention specifically provides a water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides, wherein the mass percentages of the components are as follows: 28-36% of PEG-40 hydrogenated castor oil, 18-26% of polyglycerol ricinoleate, 3-8% of water-soluble collagen hydrolyzed peptides, and the remaining mass is supplemented with GTCC (caprylic / capric triglyceride) to a mass of 100% of the water-in-oil nanoemulsion.

[0008] Preferably, the mass percentage of PEG-40 hydrogenated castor oil is 32% to 36%, specifically any value selected from 32%, 32.5%, 33%, 33.5%, 34%, 34.5%, 35%, 35.5%, 36% or any range formed by them.

[0009] Preferably, the mass percentage of polyglycerol ricinoleate is 20% to 24%, specifically any value selected from 20%, 20.5%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, 24% or any range formed by them.

[0010] Preferably, the mass percentage of the water-soluble glue protein hydrolyzed peptide is 6-8%, specifically any value selected from 6%, 6.5%, 7%, 7.5%, 8% or any range formed by them.

[0011] The present invention also provides a method for preparing the water-in-oil nanoemulsion, which specifically comprises the following steps: PEG-40 hydrogenated castor oil and polyglycerol ricinoleate are stirred evenly at low energy, GTCC (caprylic acid / capric acid triglyceride) is added to the uniform phase, stirred at low energy at room temperature, and after mixing, water-soluble collagen hydrolyzed peptide is added and stirred at low energy at room temperature to obtain the water-in-oil nanoemulsion, wherein the mass of the added PEG-40 hydrogenated castor oil is 28-36% of the mass of the water-in-oil nanoemulsion, the mass of the polyglycerol ricinoleate is 18-26% of the mass of the water-in-oil nanoemulsion, the mass of the added water-soluble collagen hydrolyzed peptide is 3-8% of the mass of the water-in-oil nanoemulsion, and the remaining mass is supplemented with GTCC until the mass of the water-in-oil nanoemulsion is 100%.

[0012] In some embodiments, the low-energy stirring is: stirring at room temperature with a rotation speed between 300 and 800 rpm, preferably 300 to 500 rpm, more preferably 380 to 400 rpm, and most preferably 400 rpm.

[0013] In some embodiments, the mass percentage of PEG-40 hydrogenated castor oil is 32% to 36%, specifically selected from any value of 32%, 32.5%, 33%, 33.5%, 34%, 34.5%, 35%, 35.5%, 36% or any range formed by them.

[0014] In some embodiments, the mass percentage of polyglycerol ricinoleate is 20% to 24%, specifically selected from any value of 20%, 20.5%, 21%, 21.5%, 22%, 22.5%, 23%, 23.5%, 24% or any range formed by them.

[0015] In some embodiments, the mass percentage of water-soluble glue protein hydrolyzed peptides is 6% to 8%, which can be specifically selected from any value of 6%, 6.5%, 7%, 7.5%, 8% or any range formed by them.

[0016] The nanoemulsion prepared by the method of the present invention and the application of the nanoemulsion in the preparation of skin care products.

[0017] Beneficial effects of the present invention:

[0018] (1) The method for preparing the water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides of the present application uses a plant-derived emulsifier with safe ingredients, reduces the amount of chemical synthetic emulsifier used, improves biocompatibility and reduces the risk of environmental pollution.

[0019] (2) The oil-in-water nanoemulsion of the present application has a particle size of less than 100 nm. The oil-in-water nanoemulsion system is prepared by a low-energy emulsification method, which solves the oil-solubility problem of collagen hydrolyzed peptides in cosmetics, allowing the collagen hydrolyzed peptides to fully exert their moisturizing, repairing, anti-aging and other effects in cosmetics. At the same time, it reduces the cost problem of the emulsification method, providing a basis for its wide application in different fields, and has good application prospects and industrial value.

[0020] (3) The emulsion component of the present invention does not require high-energy shearing and can be suitable for low-energy stirring, thereby reducing costs and effectively maintaining the activity of collagen. DETAILED DESCRIPTION

[0021] The following examples are provided for better understanding of the present invention, but are not intended to limit the present invention. The experimental methods in the following examples are conventional methods unless otherwise specified. The experimental materials used in the following examples are purchased from conventional biochemical reagent stores unless otherwise specified.

[0022] Embodiment 1:

[0023] Add an appropriate amount of polyglycerol ricinoleate (PGPR) to a certain amount of PEG-40 hydrogenated castor oil and stir evenly at 400 rpm. Add an appropriate amount of GTCC to the above homogeneous phase and stir at 400 rpm at room temperature. After mixing, add an appropriate amount of collagen hydrolyzed peptide aqueous solution and stir at room temperature at 400 rpm to obtain an oil-in-water nanoemulsion.

[0024] Table 1: Ratio of formula excipients

[0025]

[0026] The results are shown in Table 1. The emulsions formed by the three ratios of Example 1 are uniform and transparent. The particle size is less than 100 nm, the peak area at 50 nm accounts for 90%, and the peak area at 2-3 nm accounts for 10%.

[0027] Comparative Example 1:

[0028] Add an appropriate amount of polyglycerol ricinoleate to a certain amount of PEG-40 hydrogenated castor oil and stir evenly at 400 rpm. Add an appropriate amount of camellia seed oil to the above homogeneous phase and stir at 400 rpm at room temperature. After mixing, add an appropriate amount of collagen hydrolyzed peptide aqueous solution and stir at room temperature at a certain speed to obtain an oil-in-water nanoemulsion.

[0029] Table 2: Formulation excipient screening

[0030]

[0031] According to the data in Table 2, if GTCC is replaced with camellia seed oil, the formula has a translucent appearance, indicating that the nanoemulsion is not successfully formed.

[0032] Comparative Example 2:

[0033] Add an appropriate amount of polyglycerol-6 polyricinoleate to a certain amount of PEG-40 hydrogenated castor oil and stir evenly at 400 rpm (800 rpm for schemes 2 and 4), add an appropriate amount of GTCC to the above homogeneous phase, stir at 400 rpm at room temperature (800 rpm for schemes 2 and 4), add an appropriate amount of collagen hydrolyzed peptide aqueous solution after mixing, stir at 400 rpm at room temperature (800 rpm for schemes 2 and 4), and obtain an oil-in-water nanoemulsion.

[0034] Table 3: Formulation excipient screening

[0035]

[0036] According to the data in Table 3, if polyglycerol ricinoleate is replaced with polyglycerol-6 polyricinoleate, the materials in the formula will agglomerate, indicating that the nanoemulsion is not successfully formed.

[0037] Comparative Example 3:

[0038] Add an appropriate amount of polyglycerol ricinoleate to a certain amount of polyglycerol-10 laurate and stir evenly at 400 rpm (800 rpm for schemes 2 and 4), add an appropriate amount of GTCC to the above homogeneous phase, stir at 400 rpm at room temperature (800 rpm for schemes 2 and 4), add an appropriate amount of collagen hydrolyzed peptide aqueous solution after mixing, stir at 400 rpm at room temperature (800 rpm for schemes 2 and 4), and obtain an oil-in-water nanoemulsion.

[0039] Table 4: Formulation excipient screening

[0040]

[0041] According to the data in Table 4, if PEG-40 hydrogenated castor oil is replaced with polyglyceryl-10 laurate, nanoemulsion is not successfully formed.

[0042] According to the results of Example 1 and Comparative Examples 1-3, it can be seen that the collagen hydrolyzed peptide aqueous solution nanoemulsion has strict selection of emulsifiers. The formula appearance of PEG-40 hydrogenated castor oil, polyglycerol ricinoleate and GTCC in Example 1 is relatively uniform and transparent, and multiple ratios can be achieved. However, if any of the auxiliary materials is changed, it will cause a significant impact. Even after adjusting the emulsifier ratio and stirring speed, the appearance of the formula is still not greatly improved. It can be seen that the auxiliary materials described in the present invention can effectively realize the preparation of the collagen hydrolyzed peptide aqueous solution nanoemulsion, and the emulsifier ratio in this formula can be appropriately adjusted, and the appearance is uniform and transparent.

[0043] Embodiment 2:

[0044] 2.1 Add 200g of polyglycerol ricinoleate to 320g of PEG-40 hydrogenated castor oil and stir evenly. Add 400g of GTCC to the above homogeneous phase and stir at room temperature. After mixing, add 80g of collagen hydrolyzed peptide aqueous solution and stir at room temperature between 300 and 800rpm to obtain a water-in-oil nanoemulsion.

[0045] 2.2 The results in Table 2 show that in the process of preparing water-in-oil nanoemulsion, the rotation speed of 300-800 described in the present invention can achieve the preparation of nanoemulsion. However, since the components in the system have a slight viscosity, too low a rotation speed will cause the mixing time to be too long, and too high a rotation speed will cause bubbles to be generated after the system is mixed, thereby increasing the additional defoaming time, but has no effect on the formation of the nanoemulsion system. Therefore, the preferred stirring speed is 380-400rpm, and the resulting system is uniform and has fewer bubbles.

[0046] Table 2: Stirring speed optimization

[0047]

[0048] Embodiment 3:

[0049] 3.1 Add 200g of polyglycerol ricinoleate to 200-400g of PEG-40 hydrogenated castor oil and stir evenly, add 480-320g of GTCC to the above homogeneous phase, stir at room temperature, add 80g of collagen hydrolyzed peptide aqueous solution after mixing, stir at room temperature at 400rpm to obtain a water-in-oil nanoemulsion.

[0050] 3.2 The results in Table 3 show that PEG-40 hydrogenated castor oil is an emulsifier. Adding too little will cause stratification of the system after accelerated aging at 45°C for one week, and adding too much will cause flocs to form at the bottom of the system after accelerated aging at 45°C for one week. The addition ratio of the present invention is 28% to 36%. The appearance of the obtained system is still very stable after high temperature pressurization for one week, and no flocs are produced.

[0051] Table 3: Optimization of the addition ratio of PEG-40 hydrogenated castor oil

[0052]

[0053] Embodiment 4:

[0054] 4.1 Add 100-400g of polyglycerol ricinoleate to 320g of PEG-40 hydrogenated castor oil and stir evenly. Add 500-200g of GTCC to the above homogeneous phase and stir at room temperature. After mixing, add 80g of collagen hydrolyzed peptide aqueous solution and stir at room temperature at 400rpm to obtain a water-in-oil nanoemulsion.

[0055] 4.2 The results in Table 4 show that polyglycerol ricinoleate is an emulsifier. Adding too little will cause stratification of the system after accelerated aging at 45°C for one week, while adding too much will cause flocs to form at the bottom of the system after accelerated aging at 45°C for one week. The addition ratio of the present invention is 18-26%, and the appearance of the obtained system is still very stable after high temperature pressurization for one week, and no flocs are produced.

[0056] Table 4: Optimization of the addition ratio of polyglycerol ricinoleate

[0057]

[0058] Embodiment 5:

[0059] 5.1 Add 200g of polyglycerol ricinoleate to 320g of PEG-40 hydrogenated castor oil and stir evenly. Add 450-380g of GTCC to the above homogeneous phase and stir at room temperature. After mixing, add 30-100g of collagen hydrolyzed peptide aqueous solution and stir at room temperature at 400rpm to obtain a water-in-oil nanoemulsion.

[0060] 5.2 The results in Table 5 show that the aqueous solution of collagen hydrolyzed peptides belongs to the water phase. Adding too little will have no effect on the appearance of the system after accelerated aging at 45°C for one week, but will limit the loading amount of collagen hydrolyzed peptides. Adding too much will cause stratification of the system after accelerated aging at 45°C for one week. The addition ratio of the present invention is 3% to 8%. The appearance of the obtained system is still very stable after high temperature pressurization for one week, and no flocs are produced.

[0061] Table 5: Optimization of the addition ratio of collagen hydrolyzed peptide aqueous solution

[0062]

[0063] Embodiment 6:

[0064] 6.1 Sample moisturizing performance test: Samples A, B, C, D, E, and F were prepared according to the ratio in Table 6, wherein sample A was pure water, sample B was a collagen hydrolyzed peptide aqueous solution, sample C was a nanoemulsion blank matrix (i.e., the remaining components of the collagen hydrolyzed peptide were removed), sample D was a nanoemulsion containing collagen hydrolyzed peptides, i.e., the nanoemulsion prepared in ratio 1 in Example 1, sample E was an essence oil blank matrix, and sample F was an essence oil containing 2% of the nanoemulsion in ratio 1 in Example 1 (the specific product formula is shown in Table 6); 20 volunteers aged 18 to 60 were selected, the skin was first washed with pure water, then dried and left to stand for 10 minutes, and the water content of the blank skin was tested. Then, samples A, B, C, D, E, and F in Table 6 were dropped on the corresponding skin, and the skin water content of the corresponding samples was tested at 0 min, 10 min, and 60 min after slight massage for absorption. Turn on the Real Bubee Skin Moisture Tester, wait for the display to light up and hear the "beep" prompt sound, then align the probe vertically to the test area and gently stick it on the skin. After hearing the "beep" sound, the display will show the skin moisture value. Before each test, use a soft cloth or cotton paper dipped in alcohol to gently wipe and clean the probe.

[0065] Table 6: Product formula

[0066]

[0067] 6.2 The results showed that 60 minutes after application, the skin moisture content of site A was almost the same as that of the unapplied site over time, the moisture content of site B increased by 0.7% (p < 0.05) compared with site A, the moisture content of site D increased by 1% (p < 0.05) compared with site C, and the moisture content of site F increased by 2.98% (p < 0.05) compared with site E. The use of a nanoemulsion system containing collagen hydrolyzed peptides can increase the moisture content of the skin at the application site.

[0068] The average water content after 60 minutes of application is shown in Table 7. From the results in Table 7, it can be seen that the skin water content of site A is almost the same as that of the unapplied site as time goes by, the water content of site B increases by 1.67% compared with site A, the water content of site D increases by 2.67% compared with site C, and the water content of site F increases by 3.02% compared with site E. It also shows that the use of a nanoemulsion system containing collagen hydrolyzed peptides can increase the water content of the skin at the application site.

[0069] Table 7: Skin moisture content test

[0070]

[0071] Embodiment 7:

[0072] 7.1 Skin irritation test: Samples A, B, C, D, E, and F were prepared according to the ratios in Table 6 of Example 6. Twenty volunteers aged 18 to 60 were selected to conduct an irritation test with reference to the "Technical Specification for Safety of Cosmetics 2015 Edition" - Human Skin Patch Test. Samples A, B, C, D, E, and F were added to the patch tester, and then the patch tester with the samples was applied to the back of the subject with a non-irritating tape. The palm of the hand was gently pressed to evenly apply it to the skin. After 24 hours, the test patch tester was removed, and the test substance residue on the test site was gently wiped off with a moistened absorbent cotton ball. After 0.5 hours, the skin reaction was observed after the indentation disappeared. If the result was negative, it was observed again 24 hours after the patch was removed.

[0073] 7.2 The results in Table 8 show that no skin irritation occurred in 20 volunteers, regardless of whether the product containing collagen hydrolyzed peptides or the nanoemulsion composition prepared in Ratio 1 in Example 1 was used, indicating that the product is safe.

[0074] Table 8: Skin irritation results

[0075]

[0076]

Claims

1. A water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides, characterized in that: The mass percentage of each component is as follows: PEG-40 hydrogenated castor oil 28-36%, polyglycerol ricinoleate 18-26%, water-soluble glue protein hydrolyzed peptide 3-8%, and the remaining mass is supplemented with GTCC to make the mass of the water-in-oil nanoemulsion 100%.

2. The water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides according to claim 1, characterized in that: The mass percentage of PEG-40 hydrogenated castor oil is 32% to 36%.

3. The water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides according to claim 1, characterized in that: The mass percentage of polyglycerol ricinoleate is 20% to 24%.

4. The water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides according to claim 1, characterized in that: The mass percentage of the water-soluble glue protein hydrolyzed peptide is 6-8%.

5. A method for preparing a water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides, characterized in that: Specifically, PEG-40 hydrogenated castor oil and polyglycerol ricinoleate are stirred evenly at low energy, GTCC (caprylic acid / capric acid triglyceride) is added to the above homogeneous phase, stirred at low energy at room temperature, and after mixing, water-soluble collagen hydrolyzed peptide is added and stirred at low energy at room temperature to obtain an oil-in-water nanoemulsion, wherein the mass of the added PEG-40 hydrogenated castor oil is 28-36% of the mass of the oil-in-water nanoemulsion, the mass of the polyglycerol ricinoleate is 18-26% of the mass of the oil-in-water nanoemulsion, the mass of the added water-soluble collagen hydrolyzed peptide is 3-8% of the mass of the oil-in-water nanoemulsion, and the remaining mass is supplemented with GTCC until the mass of the oil-in-water nanoemulsion is 100%.

6. The preparation method according to claim 5, characterized in that: The low-energy stirring is: stirring at room temperature with a rotation speed between 300 and 800 rpm, preferably 300 to 500 rpm, more preferably 380 to 400 rpm, and most preferably 400 rpm.

7. The preparation method according to claim 5, characterized in that: The mass percentage of PEG-40 hydrogenated castor oil is 32% to 36%.

8. The preparation method according to claim 5, characterized in that: The mass percentage of polyglycerol ricinoleate is 20% to 24%.

9. The preparation method according to claim 5, characterized in that: The mass percentage of the water-soluble glue protein hydrolyzed peptide is 6% to 8%.

10. Use of the water-in-oil nanoemulsion loaded with collagen hydrolyzed peptides according to any one of claims 1 to 4 in the preparation of skin care products.

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