A composition containing recombinant collagen and its application in medical cosmetology

Abstract

The present invention relates to the field of medical cosmetology technology, and specifically to a composition containing recombinant collagen and its application in medical cosmetology. The composition containing recombinant collagen combines recombinant collagen with other active ingredients, and the components play an effective synergistic role. The overall composition of the composition is streamlined and simple in ingredients, which can effectively avoid allergies in consumers during use and meet consumers' needs for skin care and beauty.

Description

Technical Field

[0001] The present invention relates to the field of medical cosmetology technology, and in particular to a composition containing recombinant collagen and its application in medical cosmetology. Background Art

[0002] Collagen is the main extracellular matrix of the human body, accounting for about 85% of the extracellular matrix. It is the main structural protein of human tissues and organs, accounting for about 30% to 40% of the total protein in the human body. It is commonly found in the skin, blood vessels, tendons, fascia and other parts, and plays a variety of important biological functions.

[0003] At present, the collagen on the market is mainly recombinant collagen and traditional animal-derived collagen. Compared with animal-derived collagen, recombinant collagen has a single molecule and clear composition, and can obtain high-purity products; it has no virus risks, low cytotoxicity, and extremely low immunogenicity, and it is highly processable, does not require cold chain transportation, is easy to store, and has a green and environmentally friendly production and preparation process.

[0004] Recombinant collagen is a collagen produced by taking the amino acid sequence of collagen in the organism as a template, rationally designing, enzymatically cleaving and splicing it, and then transferring it into engineered cells, mainly Escherichia coli and Pichia pastoris, with the help of synthetic biology technology, and utilizing the rapid production capacity of the engineered cells. For example, the Chinese invention patent with document number CN113520899B discloses a recombinant collagen product for repairing skin photodamage. The recombinant collagen has a new protein sequence, which has a good repair effect on UV-damaged skin, has a good adhesion effect on fibroblasts, and can promote the proliferation of skin fibroblasts. It is non-irritating to the skin and has good safety.

[0005] Since recombinant collagen helps to repair damaged tissues, promote wound healing, and has an auxiliary therapeutic effect on the treatment of diseases such as arthritis, recombinant collagen has been widely used in the medical field. In addition, recombinant collagen can also improve skin elasticity and firmness, reduce the appearance of wrinkles and fine lines, and is widely used in the fields of beauty and health food production. For example, the Chinese invention patent with document number CN110934766B discloses a wrinkle-removing essence containing recombinant collagen. The wrinkle-removing essence uses recombinant collagen in combination with sodium hyaluronate, polyglutamic acid, etc., which can better exert the wrinkle-removing effect of recombinant collagen while moisturizing and locking in moisture. The Chinese invention patent with document number CN114259419B discloses a composite composition containing recombinant collagen, its preparation method and application. In the composite composition, hyaluronic acid, trehalose and recombinant collagen components are added at the same time. During the freeze-drying process, not only the molecular structure of the recombinant collagen is effectively protected from being destroyed and its biological activity is maintained for a long time, but also hyaluronic acid and trehalose themselves have excellent moisturizing effects, moisturizing skin cells, and effectively protecting DNA from damage caused by radiation. The combination of the three can synergistically exert the effects of each active ingredient. Based on the research that has been found in this type of research, recombinant collagen can play a synergistic role with other active ingredients, and the development of similar skin care products is also a current research hotspot.

[0006] Document number: CN115317429B The Chinese invention patent discloses a composition containing recombinant collagen with repairing and soothing effects, an eye cream containing the composition, and a preparation method and application. The composition includes the following preparation raw materials: 1-10 parts of recombinant collagen, 1-10 parts of sodium hyaluronate, 1-5 parts of bifid yeast fermentation product lysate, 1-15 parts of glycerol glucoside, 1-10 parts of bisabolol, 1-15 parts of eight treasures essence stock solution, and 1-15 parts of pine red plum composite essence. In the technical scheme of the invention patent, the eight treasures essence stock solution includes Chuanxiong root extract, Poria sclerotium extract, Atractylodes root extract, Bletilla striata root extract, Ampelopsis root extract, artificial bezoar, pearl extract and borneol. The composition has good skin permeability, not only has significant effects of repairing skin barrier and soothing skin inflammation, but also has the advantages of low irritation and high safety. However, the composition is complex in composition, resulting in high cost, and it is easy to cause skin allergies during use. Therefore, the development of recombinant collagen skin care products also needs to simplify the ingredients as much as possible while ensuring the application effect. Summary of the invention

[0007] In order to solve the above technical problems, the purpose of the present invention is to provide a composition containing recombinant collagen and its application in medical cosmetology. The recombinant collagen is combined with other active ingredients, and the components have an effective synergistic effect. The overall composition of the composition is streamlined and the ingredients are simple, which can effectively avoid allergies in consumers during use and meet consumers' needs for skin care and beauty.

[0008] In order to achieve the above technical effects, the present invention adopts the following technical solutions:

[0009] In a first aspect, the present invention provides a composition containing recombinant collagen, comprising the following raw material components in parts by weight: 5 to 8 parts of recombinant collagen, 1 to 3 parts of blue copper peptide, and 2 to 10 parts of Selaginella tamariscina extract.

[0010] Furthermore, the composition also includes the following raw material components in parts by weight: 0.5 to 25 parts of magnolia alcohol.

[0011] Furthermore, it comprises the following raw material components in parts by weight: 5 to 8 parts of recombinant collagen, 1 to 3 parts of blue copper peptide, 5 to 8 parts of Selaginella tamariscina extract and 1 to 20 parts of magnolia alcohol.

[0012] In a second aspect, the present invention also provides a use of the above-mentioned composition containing recombinant collagen in the preparation of medical cosmetic products.

[0013] Preferably, the application is anti-aging application.

[0014] Preferably, the application is: using the composition provided in the first aspect and auxiliary materials to prepare skin care products or cosmetics.

[0015] Preferably, the application is: in the skin care product or cosmetics, the addition amount of the composition is 0.05 wt%-5 wt%.

[0016] Preferably, the auxiliary material is selected from one or more of a moisturizer, an emollient, an emulsifier, a thickener, a preservative, a pH regulator, an antioxidant or deionized water.

[0017] Preferably, the moisturizer is any one or more of sodium hyaluronate, glycerin or ceramide; the emollient is any one or more of vaseline, lanolin or squalane; the emulsifier is any one or more of glyceryl stearate, polysorbate-20 or ceteareth-20; the thickener is any one or more of carbomer, hydroxyethyl cellulose or xanthan gum; the preservative is any one or more of phenoxyethanol, methylisothiazolinone or potassium sorbate; the pH adjuster is any one or more of citric acid, triethanolamine or sodium hydroxide; the antioxidant is any one or more of tocopherol, butylated hydroxyanisole or ascorbic acid.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The present invention provides a composition containing recombinant collagen, which is combined with recombinant collagen, blue copper peptide, and Selaginella tamariscina extract, wherein recombinant collagen can replenish collagen lost in the skin and help repair damaged skin tissue, and blue copper peptide (GHK-Cu) is a tripeptide composed of glycine-histidine-lysine, which has a significant anti-aging effect. The inventor further makes the composition play a synergistic role in skin care and beauty through the compounding of the three. In addition, on the basis of the aforementioned composition, the inventor further adds magnolia alcohol to the composition, and combines recombinant collagen, 1 to 3 parts of blue copper peptide, Selaginella tamariscina extract and magnolia alcohol together to further play a synergistic role. This synergistic effect can provide a more comprehensive skin care effect, solve skin problems from multiple angles, and comprehensively improve skin conditions, such as dryness, sagging, wrinkles, etc.

[0020] In summary, the composition provided by the present invention has a concise composition, can effectively prevent consumers from experiencing allergic reactions during use, and can enable the components of the composition to play a synergistic role to meet consumers' needs for skin care and beauty. DETAILED DESCRIPTION

[0021] The following examples are only used to more clearly illustrate the technical scheme of the present invention, and are therefore only used as examples, and cannot be used to limit the scope of the present invention. For those skilled in the art, any equivalent modifications and substitutions to the embodiments described below are also within the scope of the present invention. Therefore, the equalization transformations and modifications made without departing from the spirit and scope of the present invention should all be included in the scope of the present invention. Those who do not specify specific conditions in the embodiments are carried out according to normal conditions or the conditions recommended by the manufacturer. All reagents or instruments that do not specify manufacturers are conventional products that can be purchased commercially.

[0022] In order to better illustrate the present invention, numerous specific details are provided in the following specific embodiments. It should be understood by those skilled in the art that the present invention can be implemented without certain specific details. In other embodiments, methods, means, equipment and steps well known to those skilled in the art are not described in detail in order to highlight the subject matter of the present invention.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those commonly understood by those skilled in the art. Unless otherwise specified, the units used in this specification are all international standard units, and the numerical values ​​and numerical ranges appearing in the present invention should be understood to include the inevitable errors in industrial production.

[0024] Example 1

[0025] This embodiment provides a composition containing recombinant collagen, which includes recombinant collagen, blue copper peptide and Selaginella tamariscina extract, wherein the Selaginella tamariscina extract is prepared according to the following method:

[0026] Using 75% ethanol solution as the extraction solvent, Selaginella tamariscina was crushed and mixed with ethanol solution at a solid-liquid ratio of 1:30. Reflux extraction was adopted. The extraction time was 2 to 2.5 hours and the extraction was performed twice. After the extraction was completed, the filtrate was filtered and the filtrate was concentrated under reduced pressure and dried to obtain the Selaginella tamariscina extract.

[0027] This embodiment provides the following five specific compositions (compositions 1 to 5), as shown in Table 1:

[0028] Table 1 Compositions of compositions 1 to 5 (parts by mass)

[0029] Recombinant collagen Blue Copper Peptide Selaginella tamariscina extract Composition 1 12 2 0 Composition 2 0 2 12 Composition 3 7 2 5 Composition 4 6 2 6 Composition 5 5 2 7

[0030] Test Example 1

[0031] This test example tests the efficacy of compositions 1 to 5 in the above-mentioned embodiment 1. The specific experimental methods and experimental results are as follows:

[0032] 1.1 Free radical scavenging effect test

[0033] The experiment uses DPPH (1,1-diphenyl-2-trinitrophenylhydrazine) as the detection index, and the detection principle is:

[0034] 1,1-Diphenyl-2-trinitrophenylhydrazine (DPPH) is a stable long-lived free radical. Its ethanol solution is dark purple and has strong absorption near 517nm. When there is a free radical scavenger, the light absorption of DPPH ethanol solution is weakened due to pairing with its single electron. The degree of fading of DPPH ethanol solution is linearly related to the number of electrons it accepts, which can be used to evaluate the ability of the test sample to scavenge free radicals.

[0035] The detection method was carried out in accordance with TSHRH006-2018 "Cosmetics-Free Radical (DPPH) Scavenging Test Method", with compositions 1 to 5 as test substances, the test substance concentration was set to 50 μg / mL, and each group of experiments was repeated three times. The experimental results are shown in Table 2:

[0036] Table 2 Comparison of the removal effects of various compositions on DPPH

[0037] DPPH removal rate (%) Composition 1 64.19±1.06 Composition 2 51.81±2.29 Composition 3 82.10±3.11 Composition 4 79.21±2.49 Composition 5 78.18±2.03

[0038] The above experimental results show that among compositions 1 to 2, composition 1 has a better free radical scavenging effect. Among compositions 3 to 5, the DPPH scavenging rate can reach up to about 82% (composition 3), and only when Selaginella (Selaginella tamariscina) extract and recombinant collagen are added at the same time, a significant synergistic effect can be found between the two.

[0039] In addition, based on the above findings, the inventors further tested the relationship between the extraction method and application effect of Selaginella (Selaginella MARISCINA) extract, including:

[0040] Test group 1: The reflux extraction method in the above-mentioned Example 1 was used to extract Selaginella using 75% ethanol as the extraction solvent to obtain Selaginella tamariscina extract-ethanol extract; and composition I was prepared according to the mass ratio of recombinant collagen: blue copper peptide: Selaginella tamariscina extract-ethanol extract of 7:2:5;

[0041] Test group 2: using the same material-liquid ratio as in the above-mentioned Example 1, using water as the solvent, decoction extraction was performed, each extraction time was 2.5 hours, the extracts were extracted twice and combined, the extracts were concentrated and dried to obtain Selaginella (SELAGINELLATA MARISCINA) extract-water extract; and according to the mass ratio of the composition recombinant collagen: blue copper peptide: Selaginella (SELAGINELLATA MARISCINA) extract-water extract of 7:2:5, it was prepared as composition II;

[0042] Control group 1: Based on the test group 2, a composition III was prepared, and the composition III had the following composition:

[0043] The mass ratio of blue copper peptide: Selaginella tamariscina extract-water extract is 1:6 to prepare composition III;

[0044] The antioxidant effects of the above-mentioned composition I and composition II were tested. The test method was the same as above. The test substance concentration was set to 50 μg / mL. Each group of experiments was repeated three times. The experimental results are shown in Table 3:

[0045] Table 3 Comparison of the removal effects of composition I and composition II on DPPH

[0046] DPPH removal rate (%) Composition I 80.85±2.90 Composition II 71.32±2.34 Composition III 47.34±2.31

[0047] The above experimental results show that the effect of the combined use of Selaginella (SELAGINELLA TAMARISCINA) extract-water extract and blue copper peptide is worse than that of the combined use of Selaginella (SELAGINELLA TAMARISCINA) extract-alcohol extract and blue copper peptide, but the overall effect is not much different. However, when Selaginella (SELAGINELLA TAMARISCINA) extract, blue copper peptide and recombinant collagen are compounded, the DPPH scavenging effect of Selaginella (SELAGINELLA TAMARISCINA) extract is different whether it is extracted by water or alcohol. The synergistic effect of Selaginella (SELAGINELLA TAMARISCINA) extract-alcohol extract, blue copper peptide and recombinant collagen is more obvious. Therefore, in the aforementioned compounding method, Selaginella (SELAGINELLA TAMARISCINA) extract-alcohol extract should be selected.

[0048] 1.2 Effects on fibroblast proliferation

[0049] Fibroblasts account for more than 95% of all cells in the dermis and have the function of synthesizing and secreting collagen fibers, elastic fibers and other extracellular matrix components. This experiment evaluated the anti-aging effects of each composition by observing the effects of compositions 1 to 5 and compositions I to III on the proliferation ability of fibroblasts. The experimental principles are as follows:

[0050] Fibroblasts were cultured in a 96-well plate. The above compositions (at a concentration of 50 μg / mL) were used for intervention during the culture process. After adding each composition, the cells were incubated for 24 hours. Then, CCK-8 solution was added to each well. After culturing for 1 hour, the absorbance of each well was measured. The effect of each composition on the proliferation ability of fibroblasts was calculated based on the absorbance of each experimental group relative to that of the blank control group. The experimental results were expressed as the promotion of proliferation rate. The experimental results are shown in Tables 4 and 5:

[0051] Table 4 Effects of each composition (composition 1 to 5) on the proliferation ability of fibroblasts

[0052] Promoting proliferation rate (%) Composition 1 158.01±6.25 Composition 2 106.97±5.01 Composition 3 199.06±4.39 Composition 4 195.87±7.21 Composition 5 197.28±8.32

[0053] Table 5 Effects of each composition (composition I to III) on the proliferation ability of fibroblasts

[0054] Promoting proliferation rate (%) Composition I 201.12±9.11 Composition II 166.08±5.87 Composition III 96.31±4.19

[0055] According to the above experimental results, composition 1, that is, the combination of recombinant collagen, Selaginella (SELAGINELLA TAMARISCINA) extract and blue copper peptide, has a significant effect on the proliferation ability of fibroblasts, and the proliferation rate of fibroblasts can reach more than 195%. However, the above effect cannot be achieved when Selaginella (SELAGINELLA TAMARISCINA) extract or recombinant collagen and blue copper peptide are used alone. The experimental results show that in promoting the proliferation of fibroblasts, Selaginella (SELAGINELLA TAMARISCINA) extract and recombinant collagen have obvious synergistic effects.

[0056] According to the experimental results in Table 5, the Selaginella (SELAGINELLA TAMARISCINA) extract-water extract group (Composition III) has a large gap in the effect of promoting fibroblast proliferation compared with the Selaginella (SELAGINELLA TAMARISCINA) extract-alcohol extract group (Composition 2). In addition, in terms of synergistic effect, the synergistic effect between Selaginella (SELAGINELLA TAMARISCINA) extract-water extract and recombinant collagen is not obvious (please refer to the experimental results of Composition II and Composition 1).

[0057] In summary, the inventors found that recombinant collagen and Selaginella tamarici extract can show obvious synergistic effects, especially recombinant collagen and Selaginella tamarici extract-alcohol extract have significant synergistic effects in anti-oxidation and promoting the proliferation of fibroblasts. It is further inferred that this combination may have significant anti-wrinkle and firming effects, and is expected to play a greater role in the anti-aging field.

[0058] Example 2

[0059] Based on Example 1, this example further improves the formula based on the obvious synergistic effect between recombinant collagen and Selaginella tamariscina extract-alcohol extract that has been found. This example provides a total of 6 compositions, including compositions 6 to 11. The composition of each composition is shown in Table 6:

[0060] Table 6 Compositions of each composition (compositions 6 to 11) (parts by mass)

[0061] Recombinant collagen Blue Copper Peptide Selaginella tamariscina extract-alcohol extract Quercetin Propyl Gallate Magnolia alcohol Rutin Glabridin Composition 6 7 4 5 0 0 0 0 0 Composition 7 7 2 5 2 0 0 0 0 Composition 8 7 2 5 0 2 0 0 0 Composition 9 7 2 5 0 0 2 0 0 Composition 10 7 2 5 0 0 0 2 0 Composition 11 7 2 5 0 0 0 0 2

[0062] Test Example 2

[0063] This test example tests the efficacy of compositions 6 to 11 in the above-mentioned Example 2. The specific experimental methods and experimental results are as follows:

[0064] 2.1 Free radical scavenging effect test

[0065] The same method as in 1.1 was used for testing (the test substance concentration was 50 μg / mL). The experimental results are shown in Table 7:

[0066] Table 7 Comparison of the scavenging effects of various compositions (compositions 6 to 11) on DPPH

[0067] DPPH removal rate (%) Composition 6 84.51±2.09 Composition 7 81.32±2.30 Composition 8 79.36±4.12 Composition 9 92.41±3.37 Composition 10 73.04±1.98 Composition 11 93.01±4.06

[0068] The above experimental results show that among compositions 6 to 11, due to the addition of magnolol, glabridin and other substances, the scavenging rates of composition 9 and composition 11 on DPPH are greatly improved compared with composition 3 (DPPH scavenging rate is 82.10±3.11%), especially composition 11, which has obvious antioxidant effect. At the same time, the experimental results also show that the addition of quercetin has little effect on its antioxidant effect, and the addition of propyl gallate and rutin further affects the antioxidant effect of the original composition.

[0069] The experimental results showed that in terms of the scavenging effect on DPPH, the combination of magnolol and glycyrrhizin with recombinant collagen, blue copper peptide, and Selaginella tamariscina extract showed a good synergistic effect in the scavenging rate of DPPH.

[0070] 2.2 Effects on fibroblast proliferation

[0071] Referring to 1.2, the composition in this test example was observed to calculate the effect of each composition on the proliferation ability of fibroblasts by the absorbance value of each experimental group relative to the blank control group. The experimental results were expressed as the promotion rate of proliferation. The experimental results are shown in Table 8:

[0072] Table 8 Effects of each composition (composition 6 to 11) on the proliferation ability of fibroblasts

[0073] Promoting proliferation rate (%) Composition 6 157.01±7.98 Composition 7 138.34±6.08 Composition 8 134.02±8.99 Composition 9 245.32±7.03 Composition 10 123.01±8.18 Composition 11 189.19±8.02

[0074] According to the above experimental results, the addition of quercetin, propyl gallate and glabridin did not have a synergistic effect on the combination of recombinant collagen and Selaginella tamariscina extract, while the addition of magnolol and glabridin had a certain synergistic effect on the combination of recombinant collagen, blue copper peptide and Selaginella tamariscina extract-alcohol extract, especially the addition of magnolol, which significantly improved the effect of the original combination in promoting fibroblast proliferation.

[0075] Example 3

[0076] This embodiment provides a skin care product, which is composed of the following:

[0077] The formula of the test group product includes active ingredients and matrix ingredients, among which the active ingredients are 7 parts of recombinant collagen, 2 parts of blue copper peptide, 5 parts of Selaginella (SELAGINELLA TAMARISCINA) extract-alcohol extract, and 2 parts of magnolia alcohol; the matrix ingredients are 1 part of sodium hyaluronate, 10 parts of glycerin, 2 parts of squalane, 2 parts of cetearyl alcohol, 0.2 parts of methylparaben, and water makes up the balance to 120 parts.

[0078] The preparation method is as follows:

[0079] S1. Preparation of aqueous phase: Heat an appropriate amount of water to 70°C to 80°C, add sodium hyaluronate, and stir until completely dissolved to obtain an aqueous phase;

[0080] S2, oil phase preparation: glycerin, squalane and cetearyl alcohol are mixed evenly, and heated to 70° C. to 80° C. to obtain an oil phase;

[0081] S3. Mixing and emulsification: slowly add the oil phase into the water phase, stirring while adding, and control the stirring speed to 800-1200 rpm. Continue stirring for 10-15 minutes to fully mix and emulsify the oil phase and the water phase to form a uniform emulsion.

[0082] S4. Cooling: When the temperature of the emulsion drops to 40-45°C, add recombinant collagen, blue copper peptide, Selaginella tamariscina extract and magnolia alcohol in sequence, and continue stirring for 5-10 minutes to evenly disperse them in the emulsion. Then adjust the pH of the system to 6.5, add the preservative methylparaben, stir evenly, cool to room temperature and package.

[0083] Based on the above skin care products, the inventors further conducted efficacy tests on the aforementioned skin care products, and the test products included:

[0084] Test group products and control group products;

[0085] The matrix components in the control group product are the same as those in the test group, and the active ingredients are 7 parts of recombinant collagen, 4 parts of blue copper peptide, and 5 parts of Selaginella tamariscina extract-alcohol extract;

[0086] The test method is: 20 volunteers are recruited in each group, aged 35 to 45 years old. The two groups of volunteers use the test group products and the control group products once after cleansing in the morning and evening respectively, and the skin condition is tested before use and 28 days after use. The test results are shown in Table 9:

[0087] Table 9 Comparison of human efficacy tests between the test group and the control group

[0088]

[0089] The results of this embodiment show that different active ingredient compositions have shown good effects of improving skin condition and firming and anti-aging in human efficacy tests. Further adding magnolia alcohol on the basis of recombinant collagen, blue copper peptide and Selaginella (SELAGINELLATA MARISCINA) extract has more obvious firming and anti-aging effects, and does not cause allergic reactions during use. The above experimental results show that both compositions have good market application prospects.

[0090] The above embodiments are only used to illustrate the technical solutions of the present invention and are not intended to limit the present invention. Although the present invention is described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the purpose and scope of the technical solutions of the present invention, which should be included in the scope of the claims of the present invention. The techniques, shapes, and structural parts not described in detail in the present invention are all known technologies.

Claims

1. An anti-aging composition containing recombinant collagen, characterized in that: The invention comprises the following raw material components in parts by weight: 5 to 8 parts of recombinant collagen, 1 to 3 parts of blue copper peptide, 5 to 8 parts of Selaginella tamariscina extract and 1 to 20 parts of magnolia alcohol.

2. Use of an anti-aging composition containing recombinant collagen as claimed in claim 1 in the preparation of medical cosmetic products.

3. The use according to claim 2, characterized in that The application is anti-aging application.

4. The use according to claim 2, characterized in that The application is: using the anti-aging composition containing recombinant collagen as claimed in claim 1 and auxiliary materials to prepare cosmetics.

5. The use according to claim 4, characterized in that The application is: in the cosmetic, the added amount of the anti-aging composition containing recombinant collagen is 0.05 wt% - 5 wt%.

6. The use according to claim 4, characterized in that: The auxiliary material is selected from one or more of moisturizers, emollients, emulsifiers, thickeners, preservatives, pH regulators, antioxidants or deionized water.

7. The use according to claim 6, characterized in that: The moisturizer is any one or more of sodium hyaluronate, glycerin or ceramide; the emollient is any one or more of vaseline, lanolin or squalane; the emulsifier is any one or more of glyceryl stearate, polysorbate-20 or ceteareth-20; the thickener is any one or more of carbomer, hydroxyethyl cellulose or xanthan gum; the preservative is any one or more of phenoxyethanol, methylisothiazolinone or potassium sorbate; the pH adjuster is any one or more of citric acid, triethanolamine or sodium hydroxide; the antioxidant is any one or more of tocopherol, butylated hydroxyanisole or ascorbic acid.

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