Preparation method of oil-soluble peptide with high biocompatibility

Abstract

The invention relates to the technical field of biology, in particular to a preparation method of high-biocompatibility oil-soluble peptide. A natural ampholytic surfactant and a synthetic nonionic surfactant are compounded for use, hydrophilic polypeptides such as blue copper peptide, acetyl hexapeptide-8, conopeptide, glutathione, decarboxylated carnosine and the like are helped to be uniformly and stably dispersed in an oily solvent, and the skin permeability and activity of the polypeptides are improved.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and in particular to a method for preparing an oil-soluble peptide with high biocompatibility. Background Art

[0002] The "Catalogue of Used Cosmetic Ingredients (2021 Edition)" issued by the National Medical Products Administration includes more than 70 peptide raw materials with clear sequences, including tripeptide-1 copper (blue copper peptide), acetyl hexapeptide-8, arginine / lysine peptide (cono peptide), glutathione, decarboxylated carnosine hydrochloride, etc. These beauty peptides are the most popular type of beauty active ingredients due to their advantages such as easy absorption, clear mechanism target, low effective concentration and high safety. They are widely added to essence water, face cream, eye cream and other cosmetics. In recent years, the concept of nourishing the skin with oil has been widely spread as a new trend in skin care, but beauty peptides are difficult to add to oily formulas due to their strong hydrophilicity and low solubility in oils, which limits the application of beauty peptides in oil-based cosmetics.

[0003] In the prior art, an oil-soluble polypeptide composition is developed by fully mixing a polypeptide with a solubilizing agent such as ethylhexylglycerol, glyceryl caprylate, polyoxyethylene fatty acid ester, sorbitan laurate, polyoxyethylene oxypropylene oleate, and then adding an oily solvent. Others develop oil-soluble composite peptides by adding emulsifiers such as Tween and Span and emulsifiers / solubilizing agents such as ethylhexylglycerol and glyceryl caprylate to form microemulsion droplets in an oily solvent to encapsulate the polypeptide.

[0004] The Tween, Span, ethylhexylglycerin, glyceryl caprylate, etc. used above are all synthetic non-ionic surfactants with poor biocompatibility, which hinder the beauty peptides from penetrating into the skin to exert their activity. Summary of the invention

[0005] In view of this, the present invention provides a method for preparing a highly biocompatible oil-soluble peptide, which combines a natural amphoteric surfactant with a synthetic non-ionic surfactant to help the hydrophilic peptide to be evenly and stably dispersed in an oily solvent while increasing the skin permeability and activity of the peptide.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The present invention provides an oil-soluble peptide composition, comprising a hydrophilic polypeptide, a natural amphoteric surfactant, a nonionic surfactant, and an oily solvent;

[0008] The hydrophilic polypeptide includes at least one of blue copper peptide, acetyl hexapeptide-8, conopeptide, glutathione, and decarboxylated carnosine;

[0009] The natural amphoteric surfactant includes at least one of soybean-derived lecithin, sunflower-derived lecithin, and egg yolk-derived lecithin;

[0010] The nonionic surfactants include Span 80 and Tween 80;

[0011] The oily solvent includes at least one of squalane, cetyl ethylhexanoate, caprylic glyceryl, white oil, and olive oil.

[0012] In some specific embodiments of the present invention, the mass ratio of the hydrophilic polypeptide, the natural amphoteric surfactant, the nonionic surfactant and the oily solvent in the above-mentioned oil-soluble peptide composition is 0.032:0.1:(2.4-4.8):(14.8-17.2);

[0013] The mass ratio of Span 80 to Tween 80 is 3:1.

[0014] In some specific embodiments of the present invention, the hydrophilic polypeptide in the oil-soluble peptide composition is 0.032 g of blue copper peptide, 0.032 g of acetyl hexapeptide-8, 0.032 g of cono peptide, 0.032 g of glutathione or 0.032 g of decarboxylated carnosine;

[0015] The natural amphoteric surfactant is 0.1 g of soybean lecithin, 0.1 g of sunflower lecithin or 0.1 g of egg yolk lecithin;

[0016] The nonionic surfactant is: (i) 3.6 g of Span 80 and 1.2 g of Tween 80; or (ii) 1.8 g of Span 80 and 0.6 g of Tween 80;

[0017] The oily solvent is 14.8 g of squalane, 17.2 g of cetyl ethylhexanoate, 14.8 g of caprylic glyceryl, 17.2 g of white oil or 17.2 g of olive oil.

[0018] In some specific embodiments of the present invention, the above-mentioned oil-soluble peptide composition comprises:

[0019] (a), 0.032 g of blue copper peptide, 0.1 g of soy lecithin, 3.6 g of Span 80, 1.2 g of Tween 80 and 14.8 g of squalane; or

[0020] (b), 0.032 g of acetyl hexapeptide-8, 0.1 g of sunflower lecithin, 1.8 g of Span 80, 0.6 g of Tween 80 and 17.2 g of cetyl ethylhexanoate; or

[0021] (c), 0.032 g of cono peptide, 0.1 g of egg yolk lecithin, 3.6 g of Span 80, 1.2 g of Tween 80, and 14.8 g of caprylic acid glyceryl stearate; or

[0022] (d), 0.032 g glutathione, 0.1 g egg yolk lecithin, 1.8 g Span 80, 0.6 g Tween 80, 17.2 g white oil; or

[0023] (e), 0.032 g of decarboxylated carnosine, 0.1 g of egg yolk lecithin, 1.8 g of Span 80, 0.6 g of Tween 80, and 17.2 g of olive oil.

[0024] In some specific embodiments of the present invention, the oil-soluble peptide composition further comprises water.

[0025] The present invention also provides a method for preparing an oil-soluble peptide, comprising: mixing a natural amphoteric surfactant, a nonionic surfactant and an oily solvent, stirring, and then mixing with a hydrophilic polypeptide aqueous solution, stirring, to obtain the oil-soluble peptide;

[0026] The hydrophilic polypeptide includes at least one of blue copper peptide, acetyl hexapeptide-8, conopeptide, glutathione, and decarboxylated carnosine;

[0027] The natural amphoteric surfactant includes at least one of soybean-derived lecithin, sunflower-derived lecithin, and egg yolk-derived lecithin;

[0028] The nonionic surfactants include Span 80 and Tween 80;

[0029] The oily solvent includes at least one of squalane, cetyl ethylhexanoate, caprylic glyceryl, white oil, and olive oil.

[0030] In some specific embodiments of the present invention, the content of the hydrophilic polypeptide in the hydrophilic polypeptide aqueous solution of the above preparation method is 8 g / 100 mL;

[0031] The mass ratio of the hydrophilic polypeptide, the natural amphoteric surfactant, the nonionic surfactant and the oily solvent is 0.032:0.1:(2.4-4.8):(14.8-17.2);

[0032] The mass ratio of Span 80 to Tween 80 is 3:1.

[0033] In some specific embodiments of the present invention, the hydrophilic polypeptide in the above preparation method is blue copper peptide, acetyl hexapeptide-8, conopeptide, glutathione or decarboxylated carnosine;

[0034] The natural amphoteric surfactant is 0.1 g of soybean lecithin, 0.1 g of sunflower lecithin or 0.1 g of egg yolk lecithin;

[0035] The nonionic surfactant is: (i) 3.6 g of Span 80 and 1.2 g of Tween 80; or (ii) 1.8 g of Span 80 and 0.6 g of Tween 80;

[0036] The oily solvent is 14.8 g of squalane, 17.2 g of cetyl ethylhexanoate, 14.8 g of caprylic glyceryl, 17.2 g of white oil or 17.2 g of olive oil.

[0037] In some specific embodiments of the present invention, the above preparation method comprises:

[0038] (A), 0.1 g of soybean lecithin, 3.6 g of Span 80, 1.2 g of Tween 80 and 14.8 g of squalane are mixed, stirred, and then mixed with 0.4 mL of 8% blue copper peptide aqueous solution, stirred to obtain the oil-soluble peptide; or

[0039] (B), 0.1 g of sunflower lecithin, 1.8 g of Span 80, 0.6 g of Tween 80 and an oily solvent are mixed, stirred, and then mixed with 0.4 mL of an 8% acetyl hexapeptide-8 aqueous solution, stirred to obtain the oil-soluble peptide; or

[0040] (C), 0.1 g of egg yolk lecithin, 3.6 g of Span 80, 1.2 g of Tween 80 and an oily solvent are mixed, stirred, and then mixed with 0.4 mL of an 8% cono peptide aqueous solution, stirred to obtain the oil-soluble peptide; or

[0041] (D), 0.1 g of egg yolk lecithin, 1.8 g of Span 80, 0.6 g of Tween 80 and an oily solvent are mixed, stirred, and then mixed with 0.4 mL of an 8% glutathione aqueous solution, stirred to obtain the oil-soluble peptide; or

[0042] (E) 0.1 g of egg yolk lecithin, 1.8 g of Span 80, and 0.6 g of Tween 80 were mixed with an oily solvent, stirred, and then mixed with 0.4 mL of an 8% decarboxylated carnosine aqueous solution, stirred to obtain the oil-soluble peptide.

[0043] The present invention also provides the oil-soluble peptide prepared by the above preparation method.

[0044] The present invention has the following effects:

[0045] The oil-soluble peptide prepared by the present invention is a transparent solution with good visual effect and storage stability. The composite surfactant has multiple advantages such as strong solubilization / emulsification of non-ionic surfactants and good thermal stability of amphoteric surfactants. The use of natural amphoteric surfactants with high biocompatibility improves the skin permeability and activity of the active ingredients of the oil-soluble peptides. Experiments have shown that the essence added with natural amphoteric surfactants significantly reduces skin wrinkles and significantly improves skin roughness. Unexpectedly, when natural amphoteric surfactants are added to the essence in which the non-ionic surfactants are Span and Tween, the cumulative skin permeation is more significantly increased than that in which the non-ionic surfactants are cocamidopropyl betaine or sodium lauroyl glutamate. BRIEF DESCRIPTION OF THE DRAWINGS

[0046] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art are briefly introduced below.

[0047] Figure 1 The appearance of the blue copper peptide oil solution in Example 1 is shown;

[0048] Figure 2 The appearance of the blue copper peptide oil solution in Example 2 is shown;

[0049] Figure 3 The cumulative skin penetration of common essential oil, essential oils of comparative example 1, comparative example 2, comparative example 3, comparative example 4, embodiment 1, and embodiment 2 is shown;

[0050] Figure 4 The transdermal water loss test results of blank essential oil, essential oil containing Example 1 and Example 2 are shown;

[0051] Figure 5 The appearance of the acetyl hexapeptide-8 oil solution in Example 3 is shown;

[0052] Figure 6 The appearance of the acetyl hexapeptide-8 oil solution in Example 4 is shown;

[0053] Figure 7 The cumulative skin penetration of common essential oil, essential oils of comparative example 5, comparative example 6, comparative example 7, comparative example 8, embodiment 3, and embodiment 4 is shown;

[0054] Figure 8 The skin wrinkle reduction rates of blank essence oil, essence oil containing Example 3 and Example 4 are shown;

[0055] Fig. 9 The graph shows the skin roughness reduction rate of blank essence oil, essence oil containing Example 3 and Example 4;

[0056] Fig.10 The appearance of the conopeptide oil solution in Example 5 is shown;

[0057] Fig.11 The appearance of the conopeptide oil solution in Example 6 is shown;

[0058] Fig.12 The cumulative skin penetration of common essential oil, essential oils of Comparative Example 9, Comparative Example 10, Comparative Example 11, Comparative Example 12, Example 5, and Example 6 are shown;

[0059] Fig.13 The skin wrinkle reduction rates of blank essence oil, essence oil containing Example 5 and Example 6 are shown;

[0060] Fig.14 The graph shows the skin roughness reduction rate of blank essence oil, essence oil containing Example 5 and Example 6;

[0061] Fig.15 The appearance of glutathione oil solution in Example 7 is shown;

[0062] Fig.16 The appearance of glutathione oil solution in Example 8 is shown;

[0063] Fig.17 The cumulative skin penetration of common essential oil, essential oils of Comparative Example 13, Comparative Example 14, Comparative Example 15, Comparative Example 16, Example 7, and Example 8 are shown;

[0064] Fig.18 The skin wrinkle reduction rates of blank essence oil, essence oil containing Example 7 and Example 8 are shown;

[0065] Fig.19 The graph shows the skin roughness reduction rate of blank essence oil, essence oil containing Example 7 and Example 8;

[0066] Fig. 20 The appearance of the decarboxylated carnosine oil solution of Example 9 is shown;

[0067] Fig.21 The appearance of the decarboxylated carnosine oil solution of Example 10 is shown;

[0068] Fig. 22 The cumulative skin penetration of common essential oil, essential oils of Comparative Example 17, Comparative Example 18, Comparative Example 19, Comparative Example 20, Example 9, and Example 10 are shown;

[0069] Fig.23 The skin wrinkle reduction rates of blank essence oil, essence oil containing Example 9 and Example 10 are shown;

[0070] Fig.24 The skin roughness reduction rates of blank essence oil, essence oil containing Example 9 and essence oil containing Example 10 are shown. DETAILED DESCRIPTION

[0071] The present invention discloses a method for preparing a highly biocompatible oil-soluble peptide. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The method and application of the present invention have been described through preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the methods and applications described herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0072] The method for preparing the highly biocompatible oil-soluble peptide provided by the present invention comprises the following steps:

[0073] (1) Firstly, a hydrophilic polypeptide is dissolved in pure water to prepare an aqueous solution. The hydrophilic polypeptide is one or more of blue copper peptide, acetyl hexapeptide-8, conopeptide, glutathione, decarboxylated carnosine, etc.;

[0074] (2) adding a natural amphoteric surfactant and a nonionic surfactant to an oily solvent in a certain proportion and stirring and mixing. The natural amphoteric surfactant is lecithin, which is derived from soybean, sunflower, and egg yolk; the nonionic surfactant is a combination of Span 80 and Tween 80; and the oily solvent is one of squalane, cetyl ethylhexanoate, caprylic glyceride, white oil, olive oil, and jojoba oil;

[0075] (3) Adding the aqueous solution containing the hydrophilic polypeptide into the oil solution prepared in step (2) in a certain proportion and stirring to obtain the transparent oil solution.

[0076] It should be understood that the expression "one or more of..." includes each of the items recited after the expression individually and in various different combinations of two or more of the recited items, unless otherwise understood from the context and usage. The expression "and / or" in combination with three or more recited items should be understood to have the same meaning, unless otherwise understood from the context.

[0077] The use of the terms "comprising", "having" or "containing", including their grammatical synonyms, should generally be understood as open and non-restrictive, for example not excluding other unrecited elements or steps, unless otherwise specifically stated or otherwise understood from the context.

[0078] It should be understood that the order of steps or the order in which certain actions are performed is not important as long as the application is still operable. In addition, two or more steps or actions can be performed simultaneously.

[0079] The use of any and all examples or exemplary language such as "for example" or "including" herein is intended only to better illustrate the present application and is not intended to limit the scope of the present application. No language in this specification should be construed as indicating that any unclaimed element is essential to the practice of the present application.

[0080] In addition, the numerical ranges and parameters used to define the present application are approximate values, and the relevant values ​​in the specific embodiments have been presented as accurately as possible. However, any numerical value inherently inevitably contains standard deviations due to individual test methods. Therefore, unless otherwise expressly stated, it should be understood that all ranges, quantities, values ​​and percentages used in this disclosure are modified by "about". Here, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a specific value or range.

[0081] Unless otherwise specified, the raw materials, reagents, consumables and instruments involved in the present invention are all common commercial products and can be purchased from the market.

[0082] The present invention will be further described below in conjunction with the embodiments.

[0083] Comparative Example 1: Preparation of blue copper peptide oil solution

[0084] (1) Prepare a blue copper peptide aqueous solution: weigh 8 g of blue copper peptide hydrochloride or blue copper peptide acetate, add water to dissolve it, and then dilute to 100 mL for later use;

[0085] (2) Weigh 14.8 g of squalane and start stirring;

[0086] (3) Add 4.8 g of cocamidopropyl betaine to squalane in turn and continue stirring;

[0087] (4) Add 0.4 mL of blue copper peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain the blue copper peptide oil solution of Comparative Example 1.

[0088] Comparative Example 2: Preparation of blue copper peptide oil solution

[0089] (1) Prepare a blue copper peptide aqueous solution: weigh 8 g of blue copper peptide hydrochloride or blue copper peptide acetate, add water to dissolve it, and then dilute to 100 mL for later use;

[0090] (2) Weigh 14.8 g of squalane and start stirring;

[0091] (3) Add 4.8 g of sodium lauroyl glutamate to squalane in turn and continue stirring;

[0092] (4) Add 0.4 mL of blue copper peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain the blue copper peptide oil solution of Comparative Example 2.

[0093] Comparative Example 3: Preparation of Blue Copper Peptide Oil Solution

[0094] (1) Prepare a blue copper peptide aqueous solution: weigh 8 g of blue copper peptide hydrochloride or blue copper peptide acetate, add water to dissolve it, and then dilute to 100 mL for later use;

[0095] (2) Weigh 14.8 g of squalane and start stirring;

[0096] (3) Add 4.8 g of cocamidopropyl betaine and 0.1 g of soy lecithin to squalane in sequence and continue stirring;

[0097] (4) Add 0.4 mL of blue copper peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain the blue copper peptide oil solution of Comparative Example 1.

[0098] Comparative Example 4: Preparation of Blue Copper Peptide Oil Solution

[0099] (1) Prepare a blue copper peptide aqueous solution: weigh 8 g of blue copper peptide hydrochloride or blue copper peptide acetate, add water to dissolve it, and then dilute to 100 mL for later use;

[0100] (2) Weigh 14.8 g of squalane and start stirring;

[0101] (3) Add 4.8 g of sodium lauroyl glutamate and 0.1 g of soy lecithin to squalane in sequence and continue stirring;

[0102] (4) Add 0.4 mL of blue copper peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain the blue copper peptide oil solution of Comparative Example 2.

[0103] Example 1: Preparation of blue copper peptide oil solution

[0104] (1) Prepare a blue copper peptide aqueous solution: weigh 8 g of blue copper peptide hydrochloride or blue copper peptide acetate, add water to dissolve it, and then dilute to 100 mL for later use;

[0105] (2) Weigh 14.8 g of squalane and start stirring;

[0106] (3) Add 3.6 g of Span 80 and 1.2 g of Tween 80 to squalane in sequence and continue stirring;

[0107] (4) Add 0.4 mL of blue copper peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain a blue copper peptide oil solution without lecithin (see the appearance). Figure 1 ).

[0108] Example 2: Preparation of blue copper peptide oil solution

[0109] (1) Prepare a blue copper peptide aqueous solution: weigh 8 g of blue copper peptide hydrochloride or blue copper peptide acetate, add water to dissolve it, and then dilute to 100 mL for later use;

[0110] (2) Weigh 14.8 g of squalane and start stirring;

[0111] (3) Add 3.6 g of Span 80, 1.2 g of Tween 80, and 0.1 g of soybean lecithin to squalane in sequence and continue stirring;

[0112] (4) Add 0.4 mL of blue copper peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain a blue copper peptide oil solution containing lecithin (see the appearance). Figure 2 ).

[0113] Effect Example 1: In vitro transdermal test

[0114] (1) Take a certain amount of blue copper peptide and add it to a blank essential oil (Meadowfoam seed oil, produced by Ji'an Zhongxiang Natural Plant Co., Ltd., the same below), stir and mix well to obtain a common essential oil with a blue copper peptide mass fraction of 0.01%.

[0115] (2) At the same time, a certain amount of the blue copper peptide oil solution prepared in Comparative Example 1, Comparative Example 2, Comparative Example 3, Comparative Example 4, Example 1, and Example 2 was added to the blank essence oil and stirred to mix well, thereby obtaining the blue copper peptide essence oil of Comparative Example 1 with a blue copper peptide fraction of 0.01%, the blue copper peptide essence oil of Comparative Example 2, the blue copper peptide essence oil of Comparative Example 3, the blue copper peptide essence oil of Comparative Example 4, the blue copper peptide essence oil of Example 1 without lecithin, and the blue copper peptide essence oil of Example 2 containing lecithin.

[0116] (3) The vertical Franz diffusion cell method was used to conduct in vitro porcine skin transdermal experiments.

[0117] The back skin of the suckling pig was fixed between the receiving chamber and the supply chamber, and 1 g of the above-mentioned blue copper peptide common essence oil, blue copper peptide essence oil of comparative example 1, blue copper peptide essence oil of comparative example 2, blue copper peptide essence oil of comparative example 3, blue copper peptide essence oil of comparative example 4, blue copper peptide essence oil without lecithin in example 1, and blue copper peptide essence oil containing lecithin in example 2 were placed in the supply chamber, and physiological saline was used as the receiving liquid, and stirred and diffused at 300 r / min at 37°C. 0.5 mL of receiving liquid was taken at 2, 4, 6, 8, 10, 12 and 24 h, and an equal amount of constant temperature fresh receiving liquid was immediately added. HPLC analysis was performed to calculate the cumulative permeation of blue copper peptide at different times.

[0118] (4) Experimental results

[0119] like Figure 3 As shown, the cumulative skin penetration of blue copper peptide ordinary essence oil is 5.8mg / cm 2The cumulative skin penetration of the blue copper peptide essence oil (Comparative Example 1 and Comparative Example 2) prepared with only cocamidopropyl betaine and sodium lauroyl glutamate was 7.8 mg / cm 2 and 8.1 mg / cm 2 The cumulative skin penetration of the blue copper peptide essence oil (Comparative Example 3 and Comparative Example 4) prepared by adding lecithin to Comparative Example 1 and Comparative Example 2 was 10.2 mg / cm 2 and 13.4 mg / cm 2 ; Cumulative skin penetration of copper peptide essence oil without lecithin (Example 1): 19.3 mg / cm 2 Cumulative skin penetration of blue copper peptide essence oil containing lecithin (Example 2): 30.2 mg / cm 2 The blue copper peptide essence oil containing lecithin and the blue copper peptide essence oil without lecithin have better skin permeability than the ordinary blue copper peptide essence oil and the blue copper peptide essence oil prepared with cocamidopropyl betaine and sodium lauroyl glutamate. Among them, the blue copper peptide essence oil containing lecithin has the best skin permeability.

[0120] Effect example 2: Comparison of skin repair effects

[0121] Repair: Water loss measurement experiment

[0122] Trans Epidermal Water Loss (TEWL), also known as transdermal water loss, is a commonly used indicator to reflect the barrier function of the stratum corneum. The better the skin's protective layer, the higher the water content and the lower the TEWL value. Using TEWL as a reference indicator, the effect of liposomes in repairing the human skin barrier was evaluated.

[0123] (1) Control group: blank essential oil without blue copper peptide oil solution.

[0124] (2) Experimental group: A certain amount of the blue copper peptide oil solution prepared in Example 1 and Example 2 was added to the blank essential oil and stirred to obtain an essential oil containing lecithin and an essential oil without lecithin with a blue copper peptide fraction of 0.01%, respectively.

[0125] (3) Test method: 20 subjects aged 25 to 45 were selected, and a 3 cm × 3 cm test area was drawn on the inner side of the forearm of the subject. The skin in the test area was torn with tape and then the test sample was applied. The sample usage was 0.5 g. VapoMeter was used to test the amount of skin moisture loss before and after the skin was damaged, and 1 hour and 4 hours after the sample was applied. The test was repeated 3 times and the average value was obtained. During the experiment, the subjects were not allowed to apply any other cosmetics on the test area.

[0126] (4) Experimental results

[0127] like Figure 4 As shown, before damage, the TEWL of the skin is 10.1 g / (m 2 ·h), the TEWL of damaged skin is 25.2g / (m 2 h).

[0128] After 1 hour, the TEWL of the skin was 22.5 g / (m 2 ·h), the TEWL value of the control group was 20.5g / (m 2 ·h), the TEWL value of the blue copper peptide essence oil without lecithin was 18.4g / (m 2 h), the TEWL value of applying blue copper peptide essence oil containing lecithin is 15.3g / (m 2 h).

[0129] After 4 hours, the TEWL of the skin was 20.4 g / (m 2 ·h), the TEWL value of the control group was 18.1g / (m 2 ·h), the TEWL value of the blue copper peptide essence oil without lecithin was 14.5g / (m 2 h), the TEWL value of the blue copper peptide essence oil containing lecithin was 11.6g / (m 2 h). This shows that using blue copper peptide essence oil containing lecithin and without lecithin can better reduce skin moisture loss than ordinary blue copper peptide essence oil, among which blue copper peptide essence oil containing lecithin has the best effect.

[0130] Comparative Example 5: Preparation of Acetyl Hexapeptide-8 Oil Solution

[0131] (1) Preparation of acetyl hexapeptide-8 aqueous solution: weigh 8 g of acetyl hexapeptide-8, dissolve it in water and make up to 100 mL for later use;

[0132] (2) Weigh 17.2 g of cetyl ethylhexanoate and start stirring;

[0133] (3) Add 2.4 g of cocamidopropyl betaine to cetyl ethylhexanoate in turn and continue stirring;

[0134] (4) Add 0.4 mL of acetyl hexapeptide-8 aqueous solution to the above oily solution and continue stirring for 30 min to obtain the acetyl hexapeptide-8 oil solution of Comparative Example 3.

[0135] Comparative Example 6: Preparation of Acetyl Hexapeptide-8 Oil Solution

[0136] (1) Preparation of acetyl hexapeptide-8 aqueous solution: weigh 8 g of acetyl hexapeptide-8, dissolve it in water and make up to 100 mL for later use;

[0137] (2) Weigh 17.2 g of cetyl ethylhexanoate and start stirring;

[0138] (3) adding 2.4 g of sodium lauroyl glutamate to cetyl ethylhexanoate in sequence and continuing to stir;

[0139] (4) Add 0.4 mL of acetyl hexapeptide-8 aqueous solution to the above oily solution, and continue stirring for 30 min to obtain the acetyl hexapeptide-8 oil solution of Comparative Example 4.

[0140] Comparative Example 7: Preparation of Acetyl Hexapeptide-8 Oil Solution

[0141] (1) Preparation of acetyl hexapeptide-8 aqueous solution: weigh 8 g of acetyl hexapeptide-8, dissolve it in water and make up to 100 mL for later use;

[0142] (2) Weigh 17.2 g of cetyl ethylhexanoate and start stirring;

[0143] (3) Add 2.4 g of cocamidopropyl betaine and 0.1 g of sunflower lecithin to cetyl ethylhexanoate in sequence and continue stirring;

[0144] (4) Add 0.4 mL of acetyl hexapeptide-8 aqueous solution to the above oily solution and continue stirring for 30 min to obtain the acetyl hexapeptide-8 oil solution of Comparative Example 3.

[0145] Comparative Example 8: Preparation of Acetyl Hexapeptide-8 Oil Solution

[0146] (1) Preparation of acetyl hexapeptide-8 aqueous solution: weigh 8 g of acetyl hexapeptide-8, dissolve it in water and make up to 100 mL for later use;

[0147] (2) Weigh 17.2 g of cetyl ethylhexanoate and start stirring;

[0148] (3) Add 2.4 g of sodium lauroyl glutamate and 0.1 g of sunflower lecithin to cetyl ethylhexanoate in sequence and continue stirring;

[0149] (4) Add 0.4 mL of acetyl hexapeptide-8 aqueous solution to the above oily solution, and continue stirring for 30 min to obtain the acetyl hexapeptide-8 oil solution of Comparative Example 4.

[0150] Example 3: Preparation of Acetyl Hexapeptide-8 Oil Solution

[0151] (1) Preparation of acetyl hexapeptide-8 aqueous solution: weigh 8 g of acetyl hexapeptide-8, dissolve it in water and make up to 100 mL for later use;

[0152] (2) Weigh 17.2 g of cetyl ethylhexanoate and start stirring;

[0153] (3) Add 1.8 g of Span 80 and 0.6 g of Tween 80 to cetyl ethylhexanoate in sequence and continue stirring;

[0154] (4) Add 0.4 mL of acetyl hexapeptide-8 aqueous solution to the above oily solution and continue stirring for 30 min to obtain an acetyl hexapeptide-8 oily solution that does not contain lecithin (see the appearance). Figure 5 ).

[0155] Example 4: Preparation of Acetyl Hexapeptide-8 Oil Solution

[0156] (1) Preparation of acetyl hexapeptide-8 aqueous solution: weigh 8 g of acetyl hexapeptide-8, dissolve it in water and make up to 100 mL for later use;

[0157] (2) Weigh 17.2 g of cetyl ethylhexanoate and start stirring;

[0158] (3) Add 1.8 g of Span 80, 0.6 g of Tween 80 and 0.1 g of sunflower lecithin to cetyl ethylhexanoate in sequence and continue stirring;

[0159] (4) Add 0.4 mL of acetyl hexapeptide-8 aqueous solution to the above oily solution and continue stirring for 30 min to obtain acetyl hexapeptide-8 oil solution containing lecithin (appearance: Figure 6 ).

[0160] Effect Example 3: In vitro transdermal test

[0161] (1) Take a certain amount of acetyl hexapeptide-8, add it to the blank essential oil, stir and mix, and obtain a common essential oil with a mass fraction of acetyl hexapeptide-8 of 0.01%.

[0162] (2) At the same time, a certain amount of the acetyl hexapeptide-8 oil solution prepared in Comparative Example 5, Comparative Example 6, Comparative Example 7, Comparative Example 8, Example 3 and Example 4 was added to the blank essence oil and stirred to mix well, and the acetyl hexapeptide-8 essence oil of Comparative Example 5 with an acetyl hexapeptide-8 fraction of 0.01% was obtained, the acetyl hexapeptide-8 essence oil of Comparative Example 6, the acetyl hexapeptide-8 essence oil of Comparative Example 7, the acetyl hexapeptide-8 essence oil of Comparative Example 8, the acetyl hexapeptide-8 essence oil of Example 3 without sunflower lecithin, and the acetyl hexapeptide-8 essence oil of Example 4 containing sunflower lecithin were obtained.

[0163] (3) The vertical Franz diffusion cell method was used to conduct in vitro porcine skin transdermal experiments.

[0164] The back skin of the suckling pig was fixed between the receiving chamber and the supply chamber, and 1 g of each of the above-mentioned comparative example 5 acetyl hexapeptide-8 essence oil, comparative example 6 acetyl hexapeptide-8 essence oil, comparative example 7 acetyl hexapeptide-8 essence oil, comparative example 8 acetyl hexapeptide-8 essence oil, embodiment 3 acetyl hexapeptide-8 essence oil without sunflower lecithin, and embodiment 4 acetyl hexapeptide-8 essence oil containing sunflower lecithin was placed in the supply chamber, and physiological saline was used as the receiving liquid, and stirred and diffused at 300 r / min at 37°C. 0.5 mL of receiving liquid was taken at 2, 4, 6, 8, 10, 12 and 24 h, and an equal amount of constant temperature fresh receiving liquid was immediately supplemented. HPLC analysis was performed to calculate the cumulative permeation amount of acetyl hexapeptide-8 at different times.

[0165] (4) Experimental results

[0166] like Figure 7 As shown, the cumulative skin penetration of acetyl hexapeptide-8 normal essence oil is 6.7mg / cm 2 The cumulative skin penetration of acetyl hexapeptide-8 essence oil (Comparative Example 5 and Comparative Example 6) prepared with cocamidopropyl betaine and sodium lauroyl glutamate was 7.3 mg / cm 2 and 8.5 mg / cm 2 The cumulative skin penetration of acetyl hexapeptide-8 essence oil (Comparative Example 7 and Comparative Example 8) prepared by adding lecithin to Comparative Example 5 and Comparative Example 6 was 11.1 mg / cm 2 and 12.3 mg / cm 2 ; The cumulative skin penetration of acetyl hexapeptide-8 essence oil (Example 3) without sunflower lecithin is 18.2 mg / cm 2 ； Cumulative skin penetration of acetyl hexapeptide-8 essential oil containing sunflower lecithin (Example 4): 29.3 mg / cm 2 ; Acetyl hexapeptide-8 essence oil containing sunflower lecithin and acetyl hexapeptide-8 essence oil without sunflower lecithin has better skin permeability than ordinary acetyl hexapeptide-8 essence oil and acetyl hexapeptide-8 essence oil prepared with cocamidopropyl betaine and sodium lauroyl glutamate. Among them, acetyl hexapeptide-8 essence oil containing sunflower lecithin has the best skin permeability.

[0167] Effect example 4: Comparison of skin anti-aging effects

[0168] Anti-aging: Skin wrinkles and skin roughness analysis

[0169] (1) Control group: blank essence oil without acetyl hexapeptide-8 oil solution

[0170] (2) Experimental group: A certain amount of the acetyl hexapeptide-8 oil solution prepared in Example 3 and Example 4 was added to the blank essential oil and stirred to obtain an essential oil containing lecithin and an essential oil without lecithin with an acetyl hexapeptide-8 fraction of 0.01%, respectively.

[0171] (3) Test method: 30 subjects aged 40 to 60 were selected and randomly divided into two groups. The experimental group applied the experimental essential oil twice a day, and the control group applied the blank essential oil twice a day.

[0172] Skin wrinkle and skin roughness analyses were performed before application of the samples, and 4 and 8 weeks after application of the samples.

[0173] (4) Experimental results

[0174] like Figure 8 As shown, the number of wrinkles analyzed by VISIA-CR skin images: the control group decreased by 0.4% in 4 weeks, the experimental group using the essence oil without lecithin decreased by 2%, and the experimental group using the essence oil containing lecithin decreased by 4.0%; the control group decreased by 1.5% in 8 weeks, the experimental group using the essence oil without lecithin decreased by 6.5%, and the experimental group using the essence oil containing lecithin decreased by 10.2%. This shows that the use of acetyl hexapeptide-8 essence oil containing lecithin and acetyl hexapeptide-8 essence oil without lecithin has a better anti-wrinkle effect than acetyl hexapeptide-8 ordinary essence oil, among which the acetyl hexapeptide-8 essence oil containing lecithin has the best anti-wrinkle effect.

[0175] like Fig. 9 As shown, VisioScan VC20 Plus performed skin roughness SEr analysis: the control group decreased by 0.4%, the experimental group using the non-lecithin essence oil decreased by 3.5%, and the experimental group using the lecithin essence oil decreased by 4.8%; after 8 weeks, the control group decreased by 2%, the experimental group using the non-lecithin essence oil decreased by 5.9%, and the experimental group using the lecithin essence oil decreased by 8.7%. This shows that the use of acetyl hexapeptide-8 essence oil containing lecithin and acetyl hexapeptide-8 essence oil without lecithin has a better effect in reducing skin roughness than acetyl hexapeptide-8 ordinary essence oil, among which acetyl hexapeptide-8 essence oil containing lecithin has the best effect in reducing skin roughness.

[0176] Comparative Example 9: Preparation of Conopeptide Oil Solution

[0177] (1) Preparation of cono peptide aqueous solution: weigh 8 g of cono peptide, dissolve it in water and make up to 100 mL for later use;

[0178] (2) Weigh 14.8 g of caprylic glyceryl and start stirring;

[0179] (3) Add 4.8 g of cocamidopropyl betaine to caprylic acid glyceryl, and continue stirring;

[0180] (4) Add 0.4 mL of cono peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain the cono peptide oil solution of Comparative Example 5.

[0181] Comparative Example 10: Preparation of Conopeptide Oil Solution

[0182] (1) Preparation of cono peptide aqueous solution: weigh 8 g of cono peptide, dissolve it in water and make up to 100 mL for later use;

[0183] (2) Weigh 14.8 g of caprylic glyceryl and start stirring;

[0184] (3) Add 4.8 g of sodium lauroyl glutamate to the caprylic glyceryl esters in turn and continue stirring;

[0185] (4) Add 0.4 mL of cono peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain the cono peptide oil solution of Comparative Example 6.

[0186] Comparative Example 11: Preparation of Conopeptide Oil Solution

[0187] (1) Preparation of cono peptide aqueous solution: weigh 8 g of cono peptide, dissolve it in water and make up to 100 mL for later use;

[0188] (2) Weigh 14.8 g of caprylic glyceryl and start stirring;

[0189] (3) Add 4.8 g of cocamidopropyl betaine and 0.1 g of egg yolk lecithin to caprylic sunflower glyceride in sequence and continue stirring;

[0190] (4) Add 0.4 mL of cono peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain the cono peptide oil solution of Comparative Example 5.

[0191] Comparative Example 12: Preparation of Conopeptide Oil Solution

[0192] (1) Preparation of cono peptide aqueous solution: weigh 8 g of cono peptide, dissolve it in water and make up to 100 mL for later use;

[0193] (2) Weigh 14.8 g of caprylic glyceryl and start stirring;

[0194] (3) Add 4.8 g of sodium lauroyl glutamate and 0.1 g of egg yolk lecithin to caprylic glyceryl caprylate in sequence and continue stirring;

[0195] (4) Add 0.4 mL of cono peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain the cono peptide oil solution of Comparative Example 6.

[0196] Example 5: Preparation of conopeptide oil solution

[0197] (1) Preparation of cono peptide aqueous solution: weigh 8 g of cono peptide, dissolve it in water and make up to 100 mL for later use;

[0198] (2) Weigh 14.8 g of caprylic glyceryl and start stirring;

[0199] (3) Add 3.6 g of Span 80 and 1.2 g of Tween 80 to the caprylic acid glyceryl ester in sequence and continue stirring;

[0200] (4) Add 0.4 mL of cono peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain a cono peptide oil solution that does not contain egg yolk lecithin (see the appearance). Fig.10 ).

[0201] Example 6: Preparation of conopeptide oil solution

[0202] (1) Preparation of cono peptide aqueous solution: weigh 8 g of cono peptide, dissolve it in water and make up to 100 mL for later use;

[0203] (2) Weigh 14.8 g of caprylic glyceryl and start stirring;

[0204] (3) Add 3.6 g of Span 80, 1.2 g of Tween 80 and 0.1 g of egg yolk lecithin to caprylic acid caprylin in sequence and continue stirring;

[0205] (4) Add 0.4 mL of cono peptide aqueous solution to the above oily solution and continue stirring for 30 min to obtain a cono peptide oil solution containing egg yolk lecithin (see the appearance). Fig.11 ).

[0206] Effect Example 5: In vitro transdermal test

[0207] (1) Take a certain amount of cono peptide, add it to blank essential oil, stir and mix, and obtain a common essential oil with a cono peptide mass fraction of 0.01%.

[0208] (2) At the same time, a certain amount of the cono peptide oil solutions prepared in Comparative Example 9, Comparative Example 10, Comparative Example 11, Comparative Example 12, Example 5 and Example 6 were added to the blank essential oil and stirred to mix well, thereby obtaining the cono peptide essential oil of Comparative Example 9 with a cono peptide fraction of 0.01%, the cono peptide essential oil of Comparative Example 10, the cono peptide essential oil of Comparative Example 11, the cono peptide essential oil of Comparative Example 12, the cono peptide essential oil containing egg yolk lecithin in Example 5, and the cono peptide essential oil not containing egg yolk lecithin in Example 6.

[0209] (3) The vertical Franz diffusion cell method was used to conduct in vitro porcine skin transdermal experiments.

[0210] The back skin of the suckling pig was fixed between the receiving chamber and the supply chamber, and 1 g of the above-mentioned common cono peptide essential oil, comparative example 9 cono peptide essential oil, comparative example 10 cono peptide essential oil, comparative example 11 cono peptide essential oil, comparative example 12 cono peptide essential oil, example 5 cono peptide essential oil without egg yolk lecithin, and example 6 cono peptide essential oil containing egg yolk lecithin were taken in the supply chamber, and physiological saline was used as the receiving liquid, and stirred and diffused at 300 r / min at 37°C. 0.5 mL of receiving liquid was taken at 2, 4, 6, 8, 10, 12 and 24 h, and an equal amount of constant temperature fresh receiving liquid was immediately added. HPLC analysis was performed to calculate the cumulative penetration of cono peptide at different times.

[0211] (4) Experimental results

[0212] like Fig.12 As shown, the cumulative skin penetration of conophylline peptide ordinary essence oil is 7.4mg / cm 2 The cumulative skin penetration of the cono peptide essence oil (Comparative Example 9 and Comparative Example 10) prepared with cocamidopropyl betaine and sodium lauroyl glutamate was 8.1 mg / cm 2 and 9.2 mg / cm 2 The cumulative skin penetration of the blue copper peptide essence oil (Comparative Example 11 and Comparative Example 12) prepared by adding lecithin to Comparative Example 9 and Comparative Example 10 was 10.5 mg / cm 2 and 10.6 mg / cm 2 Cumulative skin penetration of conophylline essence oil (Example 5) without egg yolk lecithin: 17.5 mg / cm 2 ; Cumulative skin penetration of cono peptide essence oil containing egg yolk lecithin (Example 6): 34.2 mg / cm 2 The cono peptide essence oil containing egg yolk lecithin and the cono peptide essence oil without egg yolk lecithin have better skin permeability than the ordinary cono peptide essence oil and the cono peptide essence oil prepared with cocamidopropyl betaine and sodium lauroyl glutamate. Among them, the cono peptide essence oil containing lecithin has the best skin permeability.

[0213] Effect Example 6: Comparison of Skin Anti-aging Effects

[0214] Anti-aging: Skin wrinkles and skin roughness analysis

[0215] (1) Control group: blank essential oil without cono peptide oil solution

[0216] (2) Experimental group: A certain amount of the cono peptide oil solution prepared in Example 5 and Example 6 was added to the blank essential oil and stirred to obtain an essential oil containing lecithin and an essential oil without lecithin with a cono peptide fraction of 0.01%, respectively.

[0217] (3) Test method: 30 subjects aged 40 to 60 were selected and randomly divided into two groups. The experimental group applied the experimental essential oil twice a day, and the control group applied the blank essential oil twice a day.

[0218] Skin wrinkle and skin roughness analyses were performed before application of the samples, and 4 and 8 weeks after application of the samples.

[0219] (4) Experimental results

[0220] like Fig.13 As shown, the number of wrinkles analyzed by VISIA-CR skin images: the control group decreased by 0.2% in 4 weeks, the experimental group using the essence oil without lecithin decreased by 3%, and the experimental group using the essence oil containing lecithin decreased by 4.5%; the control group decreased by 2.3% in 8 weeks, the experimental group using the essence oil without lecithin decreased by 5.8%, and the experimental group using the essence oil containing lecithin decreased by 8.3%. This shows that the use of cono peptide essence oil containing lecithin and cono peptide essence oil without lecithin has a better anti-wrinkle effect than the ordinary cono peptide essence oil, and the cono peptide essence oil containing lecithin has the best anti-wrinkle effect.

[0221] like Fig.14 As shown, VisioScan VC20 Plus performed skin roughness SEr analysis: the control group decreased by 0.4%, the experimental group using the essential oil without lecithin decreased by 2.8%, and the experimental group using the essential oil containing lecithin decreased by 4.8%; after 8 weeks, the control group decreased by 2.2%, the experimental group using the essential oil without lecithin decreased by 5.1%, and the experimental group using the essential oil containing lecithin decreased by 8.1%. This shows that the use of cono peptide essential oils containing lecithin and without lecithin has a better effect in reducing skin roughness than ordinary cono peptide essential oils, among which the cono peptide essential oil containing lecithin has the best effect in reducing skin roughness.

[0222] Comparative Example 13: Preparation of glutathione oil solution

[0223] (1) Prepare glutathione aqueous solution: weigh 8 g of glutathione, dissolve it in water and make up to 100 mL for later use;

[0224] (2) Weigh 17.2 g of white oil and start stirring;

[0225] (3) Add 2.4 g of cocamidopropyl betaine to the white oil in sequence and continue stirring;

[0226] (4) Add 0.4 mL of glutathione aqueous solution to the above oily solution and continue stirring for 30 min to obtain the glutathione oil solution of Comparative Example 7.

[0227] Comparative Example 14: Preparation of glutathione oil solution

[0228] (1) Prepare glutathione aqueous solution: weigh 8 g of glutathione, dissolve it in water and make up to 100 mL for later use;

[0229] (2) Weigh 17.2 g of white oil and start stirring;

[0230] (3) Add 2.4 g of sodium lauroyl glutamate to the white oil in turn and continue stirring;

[0231] (4) Add 0.4 mL of glutathione aqueous solution to the above oily solution and continue stirring for 30 min to obtain the glutathione oil solution of Comparative Example 8.

[0232] Comparative Example 15: Preparation of glutathione oil solution

[0233] (1) Prepare glutathione aqueous solution: weigh 8 g of glutathione, dissolve it in water and make up to 100 mL for later use;

[0234] (2) Weigh 17.2 g of white oil and start stirring;

[0235] (3) Add 2.4 g of cocamidopropyl betaine and 0.1 g of egg yolk lecithin to the white oil in sequence and continue stirring;

[0236] (4) Add 0.4 mL of glutathione aqueous solution to the above oily solution and continue stirring for 30 min to obtain the glutathione oil solution of Comparative Example 7.

[0237] Comparative Example 16: Preparation of glutathione oil solution

[0238] (1) Prepare glutathione aqueous solution: weigh 8 g of glutathione, dissolve it in water and make up to 100 mL for later use;

[0239] (2) Weigh 17.2 g of white oil and start stirring;

[0240] (3) Add 2.4 g of sodium lauroyl glutamate and 0.1 g of egg yolk lecithin to the white oil in sequence and continue stirring;

[0241] (4) Add 0.4 mL of glutathione aqueous solution to the above oily solution and continue stirring for 30 min to obtain the glutathione oil solution of Comparative Example 8.

[0242] Example 7: Preparation of glutathione oil solution

[0243] (1) Prepare glutathione aqueous solution: weigh 8 g of glutathione, dissolve it in water and make up to 100 mL for later use;

[0244] (2) Weigh 17.2 g of white oil and start stirring;

[0245] (3) Add 1.8 g of Span 80 and 0.6 g of Tween 80 to the white oil in sequence and continue stirring;

[0246] (4) Add 0.4 mL of glutathione aqueous solution to the above oily solution and continue stirring for 30 min to obtain a glutathione oil solution that does not contain egg yolk lecithin (see the appearance of Fig.15 ).

[0247] Example 8: Preparation of glutathione oil solution

[0248] (1) Prepare glutathione aqueous solution: weigh 8 g of glutathione, dissolve it in water and make up to 100 mL for later use;

[0249] (2) Weigh 17.2 g of white oil and start stirring;

[0250] (3) Add 1.8 g of Span 80, 0.6 g of Tween 80 and 0.1 g of egg yolk lecithin to the white oil in sequence and continue stirring;

[0251] (4) Add 0.4 mL of glutathione aqueous solution to the above oily solution and continue stirring for 30 min to obtain a glutathione oil solution containing egg yolk lecithin (see the appearance of the solution). Fig.16 ).

[0252] Effect Example 7: In vitro transdermal test

[0253] (1) Take a certain amount of glutathione, add it to the blank essential oil, stir and mix, and obtain a common essential oil with a glutathione mass fraction of 0.01%.

[0254] (2) At the same time, a certain amount of the glutathione oil solutions prepared in Comparative Example 13, Comparative Example 14, Comparative Example 15, Comparative Example 16, Example 7, and Example 8 were added to the blank essential oil and stirred to mix well, thereby obtaining Comparative Example 13 glutathione essential oil with a glutathione fraction of 0.01%, Comparative Example 14 glutathione essential oil, Comparative Example 15 glutathione essential oil, Comparative Example 16 glutathione essential oil, Example 7 glutathione essential oil without egg yolk lecithin, and Example 8 glutathione essential oil containing egg yolk lecithin.

[0255] (3) The vertical Franz diffusion cell method was used to conduct in vitro porcine skin transdermal experiments.

[0256] The back skin of the suckling pig was fixed between the receiving chamber and the supply chamber, and 1g of each of the above-mentioned glutathione common essential oil, comparative example 13 glutathione essential oil, comparative example 14 glutathione essential oil, comparative example 15 glutathione essential oil, comparative example 16 glutathione essential oil, embodiment 7 glutathione essential oil without egg yolk lecithin, and embodiment 8 glutathione essential oil containing egg yolk lecithin was placed in the supply chamber, and physiological saline was used as the receiving liquid, and stirred and diffused at 300r / min at 37°C. 0.5mL of the receiving liquid was taken at 2, 4, 6, 8, 10, 12 and 24h, and an equal amount of constant temperature fresh receiving liquid was immediately added. HPLC analysis was performed to calculate the cumulative permeation of glutathione at different times.

[0257] (4) Experimental results

[0258] like Fig.17 As shown, the cumulative skin penetration of glutathione normal essence oil is 8.1mg / cm 2 The cumulative skin penetration of glutathione essence oil (Comparative Example 13 and Comparative Example 14) prepared with cocamidopropyl betaine and sodium lauroyl glutamate was 9.1 mg / cm 2 and 8.9 mg / cm 2 The skin cumulative penetration of the blue copper peptide essence oil (Comparative Example 15 and Comparative Example 16) prepared by adding egg yolk lecithin to Comparative Example 13 and Comparative Example 14 was 12.2 mg / cm 2 and 13.4 mg / cm 2 Cumulative skin penetration of glutathione essential oil without egg yolk lecithin (Example 7): 18.5 mg / cm 2 Cumulative skin penetration of glutathione essence oil containing egg yolk lecithin (Example 8): 34.8 mg / cm 2 Glutathione essential oils containing egg yolk lecithin and those without egg yolk lecithin have better skin permeability than ordinary glutathione essential oils and glutathione essential oils prepared with cocamidopropyl betaine and sodium lauroyl glutamate. Among them, glutathione essential oils containing egg yolk lecithin have the best skin permeability.

[0259] Effect Example 8: Comparison of Skin Anti-aging Effects

[0260] Anti-aging: Skin wrinkles and skin roughness analysis

[0261] (1) Control group: blank essential oil without glutathione oil solution

[0262] (2) Experimental group: A certain amount of the glutathione oil solution prepared in Example 7 and Example 8 was added to the blank essential oil and stirred to obtain an essential oil containing lecithin and an essential oil without lecithin with a glutathione fraction of 0.01%, respectively.

[0263] (3) Test method: 30 subjects aged 40 to 60 were selected and randomly divided into two groups. The experimental group applied the experimental essential oil twice a day, and the control group applied the blank essential oil twice a day.

[0264] Skin wrinkle and skin roughness analyses were performed before application of the samples, and 4 and 8 weeks after application of the samples.

[0265] (4) Experimental results

[0266] like Fig.18 As shown, the number of wrinkles analyzed by VISIA-CR skin images: the control group decreased by 0.3% in 4 weeks, the experimental group using the essential oil without lecithin decreased by 2.8%, and the experimental group using the essential oil containing lecithin decreased by 4.3%; the control group decreased by 2.0% in 8 weeks, the experimental group using the essential oil without lecithin decreased by 6.0%, and the experimental group using the essential oil containing lecithin decreased by 8.4%. This shows that the use of glutathione essential oils containing lecithin and lecithin has a better anti-wrinkle effect than ordinary glutathione essential oils, and the glutathione essential oil containing lecithin has the best anti-wrinkle effect.

[0267] like Fig.19 As shown, VisioScan VC20 Plus performed skin roughness SEr analysis: the control group reduced by 0.5%, the experimental group using the essential oil without lecithin reduced by 3.2%, and the experimental group using the essential oil containing lecithin reduced by 4.7%; after 8 weeks, the control group reduced by 2.5%, the experimental group using the essential oil without lecithin reduced by 5.8%, and the experimental group using the essential oil containing lecithin reduced by 8.5%. This shows that the use of glutathione essential oils containing lecithin and lecithin has a better effect in reducing skin roughness than ordinary glutathione essential oils, and the glutathione essential oil containing lecithin has the best effect in reducing skin roughness.

[0268] Comparative Example 17: Preparation of decarboxylated carnosine oil solution

[0269] (1) Preparation of decarboxylated carnosine aqueous solution: weigh 8 g of decarboxylated carnosine, add water to dissolve it and make up to 100 mL, for later use;

[0270] (2) Weigh 17.2 g of olive oil and start stirring;

[0271] (3) Add 2.4 g of cocamidopropyl betaine to the white oil in sequence and continue stirring;

[0272] (4) Add 0.4 mL of decarboxylated carnosine aqueous solution to the above oily solution, and continue stirring for 30 min to obtain the decarboxylated carnosine oil solution of Comparative Example 9.

[0273] Comparative Example 18: Preparation of decarboxylated carnosine oil solution

[0274] (1) Preparation of decarboxylated carnosine aqueous solution: weigh 8 g of decarboxylated carnosine, add water to dissolve and make up to 100 mL, for later use;

[0275] (2) Weigh 17.2 g of olive oil and start stirring;

[0276] (3) Add 2.4 g of sodium lauroyl glutamate to the white oil in turn and continue stirring;

[0277] (4) Add 0.4 mL of decarboxylated carnosine aqueous solution to the above oily solution, and continue stirring for 30 min to obtain the decarboxylated carnosine oil solution of Comparative Example 10.

[0278] Comparative Example 19: Preparation of decarboxylated carnosine oil solution

[0279] (1) Preparation of decarboxylated carnosine aqueous solution: weigh 8 g of decarboxylated carnosine, add water to dissolve it and make up to 100 mL, for later use;

[0280] (2) Weigh 17.2 g of olive oil and start stirring;

[0281] (3) Add 2.4 g of cocamidopropyl betaine and 0.1 g of egg yolk lecithin to the white oil in sequence and continue stirring;

[0282] (4) Add 0.4 mL of decarboxylated carnosine aqueous solution to the above oily solution, and continue stirring for 30 min to obtain the decarboxylated carnosine oil solution of Comparative Example 9.

[0283] Comparative Example 20: Preparation of decarboxylated carnosine oil solution

[0284] (1) Preparation of decarboxylated carnosine aqueous solution: weigh 8 g of decarboxylated carnosine, add water to dissolve it and make up to 100 mL, for later use;

[0285] (2) Weigh 17.2 g of olive oil and start stirring;

[0286] (3) Add 2.4 g of sodium lauroyl glutamate and 0.1 g of egg yolk lecithin to the white oil in sequence and continue stirring;

[0287] (4) Add 0.4 mL of decarboxylated carnosine aqueous solution to the above oily solution, and continue stirring for 30 min to obtain the decarboxylated carnosine oil solution of Comparative Example 10.

[0288] Embodiment 9: decarboxylated carnosine oil solution preparation

[0289] (1) Preparation of decarboxylated carnosine aqueous solution: weigh 8 g of decarboxylated carnosine, add water to dissolve and make up to 100 mL, for later use;

[0290] (2) Weigh 17.2 g of olive oil and start stirring;

[0291] (3) Add 1.8 g of Span 80 and 0.6 g of Tween 80 to the white oil in sequence and continue stirring;

[0292] (4) Add 0.4 mL of decarboxylated carnosine aqueous solution to the above oily solution and continue stirring for 30 min to obtain a decarboxylated carnosine oil solution free of egg yolk lecithin (see the appearance). Fig. 20 ).

[0293] Embodiment 10: decarboxylated carnosine oil solution preparation

[0294] (1) Preparation of decarboxylated carnosine aqueous solution: weigh 8 g of decarboxylated carnosine, add water to dissolve and make up to 100 mL, for later use;

[0295] (2) Weigh 17.2 g of olive oil and start stirring;

[0296] (3) Add 1.8 g of Span 80, 0.6 g of Tween 80 and 0.1 g of egg yolk lecithin to the white oil in sequence and continue stirring;

[0297] (4) Add 0.4 mL of decarboxylated carnosine aqueous solution to the above oily solution and continue stirring for 30 min to obtain a decarboxylated carnosine oil solution containing egg yolk lecithin (see the appearance). Fig.21 ).

[0298] Effect Example 9: In vitro transdermal test

[0299] (1) Take a certain amount of decarboxylated carnosine, add it to blank essential oil, stir and mix, and obtain a common essential oil with a decarboxylated carnosine mass fraction of 0.01%.

[0300] (2) At the same time, a certain amount of the decarboxylated carnosine oil solutions prepared in Comparative Example 17, Comparative Example 18, Comparative Example 19, Comparative Example 10, Example 9 and Example 10 were added to the blank essential oil and stirred to mix well, respectively obtaining the decarboxylated carnosine essential oil of Comparative Example 17 with a decarboxylated carnosine fraction of 0.01%, the decarboxylated carnosine essential oil of Comparative Example 18, the decarboxylated carnosine essential oil of Comparative Example 19, the decarboxylated carnosine essential oil of Comparative Example 20, the decarboxylated carnosine essential oil of Example 9 without egg yolk lecithin, and the decarboxylated carnosine essential oil of Example 10 containing egg yolk lecithin.

[0301] (3) The vertical Franz diffusion cell method was used to conduct in vitro porcine skin transdermal experiments.

[0302] The back skin of suckling pig was fixed between the receiving chamber and the supply chamber, and 1g of the above-mentioned decarboxylated carnosine common essential oil, comparative example 17 decarboxylated carnosine essential oil, comparative example 18 decarboxylated carnosine essential oil, comparative example 19 decarboxylated carnosine essential oil, comparative example 20 decarboxylated carnosine essential oil, embodiment 9 decarboxylated carnosine essential oil without egg yolk lecithin, and embodiment 10 decarboxylated carnosine essential oil containing egg yolk lecithin were taken into the supply chamber, and physiological saline was used as the receiving solution, and stirred and diffused at 300r / min at 37°C. 0.5mL of receiving solution was taken at 2, 4, 6, 8, 10, 12 and 24h, and an equal amount of constant temperature fresh receiving solution was immediately supplemented. HPLC analysis was performed to calculate the cumulative permeation amount of decarboxylated carnosine at different times.

[0303] (4) Experimental results

[0304] like Fig. 22 As shown in the figure, the cumulative skin penetration of decarboxylated carnosine ordinary essence oil is 9.2mg / cm 2 The cumulative skin penetration of decarboxylated carnosine essence oil (Comparative Example 17 and Comparative Example 18) prepared with cocamidopropyl betaine and sodium lauroyl glutamate was 10.3 mg / cm 2 and 12.2 mg / cm 2 The cumulative skin penetration of decarboxylated carnosine essential oils (Comparative Examples 19 and 20) prepared by adding egg yolk lecithin to Comparative Examples 17 and 18 was 13.7 mg / cm 2 and 15.6 mg / cm 2 The cumulative skin penetration of decarboxylated carnosine essence oil (Example 9) without egg yolk lecithin is 20.6 mg / cm 2 Cumulative skin penetration of decarboxylated carnosine essence oil containing egg yolk lecithin (Example 10): 36.7 mg / cm 2 This shows that decarboxylated carnosine essence oil containing lecithin and decarboxylated carnosine essence oil without lecithin has better skin permeability than ordinary decarboxylated carnosine essence oil and decarboxylated carnosine essence oil prepared with cocamidopropyl betaine and sodium lauroyl glutamate, among which decarboxylated carnosine essence oil containing egg yolk lecithin has the best skin permeability.

[0305] Effect Example 10: Comparison of Skin Anti-aging Effects

[0306] Anti-aging: Skin wrinkles and skin roughness analysis

[0307] (1) Control group: blank essence oil without decarboxylated carnosine oil solution

[0308] (2) Experimental group: A certain amount of the decarboxylated carnosine oil solution prepared in Example 9 and Example 10 was added to the blank essential oil and stirred to obtain an essential oil containing lecithin and an essential oil without lecithin and having a decarboxylated carnosine fraction of 0.01%, respectively.

[0309] (3) Test method: 30 subjects aged 40 to 60 were selected and randomly divided into two groups. The experimental group applied the experimental essential oil twice a day, and the control group applied the blank essential oil twice a day.

[0310] Skin wrinkle and skin roughness analyses were performed before application of the samples, and 4 and 8 weeks after application of the samples.

[0311] (4) Experimental results

[0312] like Fig.23 As shown, the number of wrinkles analyzed by VISIA-CR skin images: the control group decreased by 0.3% in 4 weeks, the experimental group using the essence oil without lecithin decreased by 3.2%, and the experimental group using the essence oil containing lecithin decreased by 4.5%; the control group decreased by 2.5% in 8 weeks, the experimental group using the essence oil without lecithin decreased by 7.5%, and the experimental group using the essence oil containing lecithin decreased by 9.6%. This shows that the use of decarboxylated carnosine essence oil containing lecithin and decarboxylated carnosine essence oil without lecithin has a better anti-wrinkle effect than the ordinary decarboxylated carnosine essence oil, and the decarboxylated carnosine essence oil containing lecithin has the best anti-wrinkle effect.

[0313] like Fig.24 As shown, VisioScan VC20 Plus performed skin roughness SEr analysis: the control group decreased by 0.4%, the experimental group using the essential oil without lecithin decreased by 3.6%, and the experimental group using the essential oil containing lecithin decreased by 5.3%; after 8 weeks, the control group decreased by 2.6%, the experimental group using the essential oil without lecithin decreased by 7.1%, and the experimental group using the essential oil containing lecithin decreased by 10.2%. This shows that the use of decarboxylated carnosine essential oil containing lecithin and decarboxylated carnosine essential oil has a better effect in reducing skin roughness than decarboxylated carnosine ordinary essential oil, among which the decarboxylated carnosine essential oil containing lecithin has the best effect in reducing skin roughness.

[0314] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. An oil-soluble peptide composition, characterized in that: Including hydrophilic peptides, natural amphoteric surfactants, nonionic surfactants, and oily solvents; The hydrophilic polypeptide includes at least one of blue copper peptide, acetyl hexapeptide-8, conopeptide, glutathione, and decarboxylated carnosine; The natural amphoteric surfactant includes at least one of soybean-derived lecithin, sunflower-derived lecithin, and egg yolk-derived lecithin; The nonionic surfactants include Span 80 and Tween 80; The oily solvent includes at least one of squalane, cetyl ethylhexanoate, caprylic glyceryl, white oil, and olive oil.

2. The oil-soluble peptide composition according to claim 1, characterized in that The mass ratio of the hydrophilic polypeptide, the natural amphoteric surfactant, the nonionic surfactant and the oily solvent is 0.032:0.1:(2.4-4.8):(14.8-17.2); The mass ratio of Span 80 to Tween 80 is 3:

1.

3. The oil-soluble peptide composition according to claim 1 or 2, characterized in that The hydrophilic polypeptide is 0.032 g of blue copper peptide, 0.032 g of acetyl hexapeptide-8, 0.032 g of cono peptide, 0.032 g of glutathione or 0.032 g of decarboxylated carnosine; The natural amphoteric surfactant is 0.1 g of soybean lecithin, 0.1 g of sunflower lecithin or 0.1 g of egg yolk lecithin; The nonionic surfactant is: (i) 3.6 g of Span 80 and 1.2 g of Tween 80; or (ii) 1.8 g of Span 80 and 0.6 g of Tween 80; The oily solvent is 14.8 g of squalane, 17.2 g of cetyl ethylhexanoate, 14.8 g of caprylic glyceryl, 17.2 g of white oil or 17.2 g of olive oil.

4. The oil-soluble peptide composition according to any one of claims 1 to 3, characterized in that include: (a), 0.032 g of blue copper peptide, 0.1 g of soy lecithin, 3.6 g of Span 80, 1.2 g of Tween 80 and 14.8 g of squalane; or (b), 0.032 g of acetyl hexapeptide-8, 0.1 g of sunflower lecithin, 1.8 g of Span 80, 0.6 g of Tween 80 and 17.2 g of cetyl ethylhexanoate; or (c), 0.032 g of cono peptide, 0.1 g of egg yolk lecithin, 3.6 g of Span 80, 1.2 g of Tween 80, and 14.8 g of caprylic acid glyceryl stearate; or (d), 0.032 g glutathione, 0.1 g egg yolk lecithin, 1.8 g Span 80, 0.6 g Tween 80, 17.2 g white oil; or (e), 0.032 g of decarboxylated carnosine, 0.1 g of egg yolk lecithin, 1.8 g of Span 80, 0.6 g of Tween 80, and 17.2 g of olive oil.

5. The oil-soluble peptide composition according to any one of claims 1 to 4, characterized in that Also includes water.

6. A method for preparing an oil-soluble peptide, characterized in that: include: The natural amphoteric surfactant and the nonionic surfactant are mixed with an oily solvent, and then mixed with a hydrophilic polypeptide aqueous solution, and stirred to obtain the oil-soluble peptide; The hydrophilic polypeptide includes at least one of blue copper peptide, acetyl hexapeptide-8, conopeptide, glutathione, and decarboxylated carnosine; The natural amphoteric surfactant includes at least one of soybean-derived lecithin, sunflower-derived lecithin, and egg yolk-derived lecithin; The nonionic surfactants include Span 80 and Tween 80; The oily solvent includes at least one of squalane, cetyl ethylhexanoate, caprylic glyceryl, white oil, and olive oil.

7. The preparation method according to claim 6, characterized in that: The content of the hydrophilic polypeptide in the hydrophilic polypeptide aqueous solution is 8 g / 100 mL; The mass ratio of the hydrophilic polypeptide, the natural amphoteric surfactant, the nonionic surfactant and the oily solvent is 0.032:0.1:(2.4-4.8):(14.8-17.2); The mass ratio of Span 80 to Tween 80 is 3:

1.

8. The preparation method according to claim 6 or 7, characterized in that: The hydrophilic polypeptide is blue copper peptide, acetyl hexapeptide-8, conopeptide, glutathione or decarboxylated carnosine; The natural amphoteric surfactant is 0.1 g of soybean lecithin, 0.1 g of sunflower lecithin or 0.1 g of egg yolk lecithin; The nonionic surfactant is: (i) 3.6 g of Span 80 and 1.2 g of Tween 80; or (ii) 1.8 g of Span 80 and 0.6 g of Tween 80; The oily solvent is 14.8 g of squalane, 17.2 g of cetyl ethylhexanoate, 14.8 g of caprylic glyceryl, 17.2 g of white oil or 17.2 g of olive oil.

9. The preparation method according to any one of claims 6 to 7, characterized in that: include: (A), 0.1 g of soybean lecithin, 3.6 g of Span 80, 1.2 g of Tween 80 and 14.8 g of squalane are mixed, and then mixed with 0.4 mL of 8% blue copper peptide aqueous solution, and stirred to obtain the oil-soluble peptide; or (B), 0.1 g of sunflower lecithin, 1.8 g of Span 80, 0.6 g of Tween 80 and an oily solvent are mixed, and then mixed with 0.4 mL of an 8% acetyl hexapeptide-8 aqueous solution, and stirred to obtain the oil-soluble peptide; or (C), mixing 0.1 g of egg yolk lecithin, 3.6 g of Span 80, and 1.2 g of Tween 80 with an oily solvent, and then mixing with 0.4 mL of an 8% cono peptide aqueous solution, and stirring to obtain the oil-soluble peptide; or (D), mixing 0.1 g of egg yolk lecithin, 1.8 g of Span 80, and 0.6 g of Tween 80 with an oily solvent, and then mixing with 0.4 mL of an 8% glutathione aqueous solution, and stirring to obtain the oil-soluble peptide; or (E) 0.1 g of egg yolk lecithin, 1.8 g of Span 80, and 0.6 g of Tween 80 were mixed with an oily solvent, and then mixed with 0.4 mL of an 8% decarboxylated carnosine aqueous solution, and stirred to obtain the oil-soluble peptide.

10. The oil-soluble peptide obtained by the preparation method according to any one of claims 6 to 9.

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