CN113476340A - Grafted chitosan derivative modified whitening co-delivery nano composition and preparation method and application thereof - Google Patents

Abstract

The invention belongs to the technical field of cosmetic raw materials, and discloses a grafted chitosan derivative modified whitening co-delivery nano composition, and a preparation method and application thereof. The grafted chitosan derivative modified whitening co-delivery nano composition is prepared by the following method: firstly, preparing the mixture containing phenethyl resorcinol and glycyrrhiza glabraDing, myo-peptide, V_CThe whitening composite co-delivery nanometer ethosome containing five active whitening components such as ethyl ether and nicotinamide is subjected to ion crosslinking by using the grafted chitosan derivative and the whitening composite co-delivery nanometer ethosome, so that the grafted chitosan derivative modified whitening co-delivery nanometer composition is obtained. The grafted chitosan derivative has good solubility in aqueous solution with a wider pH range, and has abundant cations, so that the grafted chitosan derivative has better adhesion, can prolong the contact time with cells, and enhances the drug absorption. The cationic whitening co-delivery nano composition modified by the grafted chitosan derivative provided by the invention can effectively improve the stability and bioavailability, and further promote the application of the composition in the field of cosmetics.

Description

Grafted chitosan derivative modified whitening co-delivery nano composition and preparation method and application thereof

Technical Field

The invention belongs to the technical field of cosmetic raw materials, and particularly relates to a grafted chitosan derivative modified whitening co-delivery nano composition, and a preparation method and application thereof.

Background

Phenethylresorcinol, chemically known as 4- (1-phenylethyl) -1, 3-benzenediol, also known as Symwhite377, is a white crystalline solid with a bland odor that is soluble in oils and glycols. The phenethyl resorcinol is derived from a natural whitening component, namely pinosylvin, in Pinus densiflora and is a novel efficient whitening active component which is similar to a dihydropinosylvin structure and formed by chemical transformation. The phenethyl resorcinol can effectively inhibit the activity of the tyrosinase, has strong antioxidant capacity, can effectively whiten the skin, improve uneven skin color and reduce skin coloration caused by ultraviolet irradiation on the skin. In 2012, the chinese national food and drug administration (SFDA) formally approved phenethylresorcinol as an ingredient for cosmetic applications.

However, the phenethyl resorcinol has the following problems in application: the solubility of the phenethyl resorcinol in water is low, the content of the phenethyl resorcinol is too low when the phenethyl resorcinol is used in a formula of a cosmetic, and the whitening effect is difficult to achieve, and the existing research shows that the phenethyl resorcinol can effectively whiten when the dosage is more than 0.5 percent, but the addition of high concentration is easy to cause the allergic reaction of skin; ② the phenethyl resorcinol has optical instability, rapid inactivation and discoloration under illumination condition, and serious influence on the stability and functional effect of the product.

In addition, glabridin is also one of components that inhibit the activity of neuraminidase efficiently, is derived from specific licorice, and is widely used for skin whitening, skin color reduction, and skin coloration reduction due to ultraviolet irradiation. Glabridin is a natural whitening agent, but has the characteristics of insolubility in water, high melting point, instability in illumination and high temperature environment, easy color decomposition and the like due to the physicochemical property; meanwhile, due to the barrier function of the skin, the transdermal permeability and the active absorption capacity of the glabridin are weak, so that the application of the glabridin is limited to a certain extent.

With the development of nanotechnology, nano-preparations (NPs) play more and more important roles in applications such as skin penetration promotion, enzyme resistance, slow release and the like. Research shows that the physicochemical properties (such as particle size, form, charged and surface characteristics) of the nano preparation play an important role in absorption, distribution, penetration and metabolism in vivo. Through the intensive study on the physicochemical properties of the nano-preparation, a more ideal drug delivery system can be designed, such as: improving the stability and bioavailability of the active substance, prolonging the long circulation in vivo, and the like. In the prior art, no report of the cationic whitening co-delivery nano composition modified by the grafted chitosan derivative exists.

Disclosure of Invention

In order to overcome the problems of poor water solubility and stability of phenylethyl resorcinol and glabridin in the prior art and how to safely and effectively absorb the phenylethyl resorcinol and the glabridin by skin, the invention mainly aims to provide a preparation method of a grafted chitosan derivative modified whitening co-delivery nano composition.

The invention also aims to provide the grafted chitosan derivative modified whitening co-delivery nano composition prepared by the method.

The invention further aims to provide application of the grafted chitosan derivative modified whitening co-delivery nano composition.

The purpose of the invention is realized by the following scheme:

a preparation method of a grafted chitosan derivative modified whitening co-delivery nano composition comprises the following steps:

(1) weighing 5-10% of phenethyl resorcinol, 1-5% of glabridin, 5-10% of 1, 2-propylene glycol and 5-10% of 1, 3-butanediol according to the mass percentage, and fully dissolving the phenethyl resorcinol and the glabridin by ultrasonic; then sequentially adding 10-20% of glycerol, 1-10% of GTCC, 1-10% of Tween-20, 1-10% of isononyl isononanoate, 1-5% of soybean lecithin and 1-5% of polyglycerol-10 oleate, and stirring at 25-30 ℃ to fully dissolve all the components to obtain an A phase;

(2) weighing 1-5% of carnosine, 1-5% of Vc ethyl ether, 1-5% of nicotinamide and 20-60% of deionized water in percentage by mass, and performing ultrasonic treatment to completely dissolve all the components to obtain a phase B;

(3) adding the phase B into the phase A at a constant speed under magnetic stirring, stirring at 25-30 deg.C to make the two phases uniformly mixed, continuing to react for 20min, and performing high-speed shearing dispersion by using high-shear homogenizing emulsifying machine to obtain crude ethosome;

(4) carrying out high-pressure homogenization on the crude alcohol body by a high-pressure micro-jet homogenization method to obtain a whitening composite co-delivery nanometer alcohol body;

(5) and (3) dissolving the grafted chitosan derivative in an aqueous solution in advance to obtain a grafted chitosan derivative solution, dripping the whitening composite co-delivery nano ethosome solution obtained in the step (4) into the grafted chitosan derivative solution at a constant speed by using a peristaltic pump under the stirring condition, and continuing stirring for 10-30min after finishing dripping to obtain the grafted chitosan derivative modified whitening co-delivery nano composition liposome.

The mass percentages in the step (1) and the step (2) refer to mass percentages of the components in the total mass.

The stirring speed in the step (3) is preferably 150-400 rpm/min; the power of the high-shear homogenizing and emulsifying machine in the step (3) is 500-.

The high-pressure homogenizing pressure in the step (4) is 1.0-1.5MPa, and the circulation is performed for 3-6 times.

The grafted chitosan derivative in the step (5) is at least one of N-alkyl chitosan, O-alkyl chitosan, lauryl-succinylated chitosan, carboxymethyl chitosan, 2-hydroxypropyl trimethyl ammonium chloride chitosan, chitosan trimethyl quaternary ammonium salt, polyacrylamide grafted chitosan, polylactic acid grafted chitosan, polycaprolactone grafted chitosan, polyvinyl alcohol grafted chitosan and the like, and preferably 2-hydroxypropyl trimethyl ammonium chloride chitosan.

The concentration of the grafted chitosan derivative in the grafted chitosan derivative solution in the step (5) is 2-16 mg/mL;

the mass ratio of the grafted chitosan derivative solution to the whitening composite co-delivery nanometer ethosome solution in the step (5) is 1: 2-1: 5, preferably 1:3 (w/w).

The stirring speed in the step (5) is 150-400rpm/min, and the dropping speed of the peristaltic pump is 20-50 rpm/min.

The grafted chitosan derivative modified whitening co-delivery nano composition prepared by the method.

The grafted chitosan derivative modified whitening co-delivery nano composition is applied to cosmetics.

The mechanism of the invention is as follows:

the liposome modified by chitosan and its derivatives can form cationic Drug Delivery Systems (PDDS), and the cationic nanocarrier has better cell fusion. The natural chitosan has low solubility in water, and the functional modification of the hydroxyl group, amino group and other groups on the structure can greatly improve the solubility of the chitosan, and can endow the chitosan with richer cations, so that the modified chitosan derivative has better adhesion, the contact time with cells can be prolonged, and the drug absorption can be enhanced. The invention provides a grafted chitosan derivative modified cationic whitening co-delivery nano composition and a preparation method thereof, which can effectively improve the stability and bioavailability and further promote the application of the composition in the field of cosmetics.

Compared with the prior art, the invention has the following advantages and beneficial effects:

(1) the invention provides a grafted chitosan derivative modified cationic whitening co-delivery nano composition, and a preparation method and application thereof. Firstly, compared with the existing anionic nano-liposome on the market, the cationic nano-liposome has better cell fusion; and secondly, the water solubility of the grafted chitosan derivative subjected to functional modification of natural chitosan is greatly improved, the grafted chitosan derivative shows good solubility in aqueous solution with a wider pH range, and has richer cations, so that the grafted chitosan derivative has better adhesion, can prolong the contact time with cells, enhances the bioavailability of drug absorption, and further promotes the application of the grafted chitosan derivative in the field of cosmetics.

(2) The invention adopts phenethyl resorcinol, glabridin, carnosine and V_CThe five active ingredients are synergistic, the activity of the neuraminidase is effectively inhibited, the generation and migration of melanin are blocked from the source, the skin is antioxidant and anti-saccharification, the immunity of the skin is improved, and the multi-effect multi-target whitening and skin brightening are realized.

Drawings

FIG. 1 is a graph of the results of testing human volunteers for skin color ITA ° values before, 2 weeks, 4 weeks, and 8 weeks after use of the emulsions of example 1, example 2, comparative example 1, comparative example 2, and the blank;

FIG. 2 is a graph of the results of testing the MI values of skin melanin in human volunteers before, after 2 weeks, 4 weeks, and 8 weeks of use of the emulsions of example 1, example 2, comparative example 1, comparative example 2, and the blank.

Detailed Description

The present invention will be described in further detail with reference to examples and drawings, but the embodiments of the present invention are not limited thereto. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The reagents used in the examples are commercially available without specific reference.

Example 1

The preparation method comprises the following steps of carrying out ultrasonic treatment on 5% of phenethyl resorcinol, 1% of glabridin, 10% of 1, 2-propylene glycol and 10% of 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% of glycerol, 5% of GTCC, 2.5% of tween-20, 2.5% of polyglycerol-10 oleate, 5% of isononyl isononanoate and 1% of soybean lecithin, dissolving the components by magnetic stirring at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 2% carnosine and 3% VC ethyl ether, 1% nicotinamide and 32% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. Preparing 8mg/mL 2-hydroxypropyl trimethyl ammonium chloride chitosan aqueous solution according to the following steps: whitening co-delivery nanoethosome solution ═ 1:3(w/w) performing ion crosslinking, dripping the whitening co-delivery nano ethosome solution into a 2-hydroxypropyl trimethyl ammonium chloride chitosan solution by using a peristaltic pump (the speed of 30 rpm/min), and stirring at the magnetic stirring speed of 200rpm/min for 30min to obtain the 2-hydroxypropyl trimethyl ammonium chloride chitosan modified whitening co-delivery nano composition liposome; the particle size of the co-delivery nano liposome modified by 2-hydroxypropyl trimethyl ammonium chloride chitosan is detected, and the co-delivery nano liposome is 97.2nm in particle size and 38.4mV in Zeta potential.

Example 2

The preparation method comprises the following steps of carrying out ultrasonic treatment on 5% of phenethyl resorcinol, 1% of glabridin, 10% of 1, 2-propylene glycol and 10% of 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% of glycerol, 5% of GTCC, 2.5% of tween-20, 2.5% of polyglycerol-10 oleate, 5% of isononyl isononanoate and 1% of soybean lecithin, dissolving the components by magnetic stirring at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 2% carnosine and 3% VC ethyl ether, 1% nicotinamide and 32% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. Preparing 8mg/mL carboxymethyl chitosan aqueous solution according to the following steps: whitening co-delivery nanoethosome solution ═ 1:3(w/w) carrying out ion crosslinking, dripping the whitening co-delivery nano ethosome solution into a carboxymethyl chitosan solution by using a peristaltic pump (the speed of 30 rpm/min), and carrying out magnetic stirring at the speed of 200rpm/min for 30min to obtain a carboxymethyl chitosan modified whitening co-delivery nano composition liposome; the particle size of the carboxymethyl chitosan modified whitening co-delivery nano liposome is detected, and the co-delivery nano liposome is obtained to be 95.8nm in particle size and 34.2mV in Zeta potential.

Example 3

The preparation method comprises the following steps of carrying out ultrasonic treatment on 5% of phenethyl resorcinol, 1% of glabridin, 10% of 1, 2-propylene glycol and 10% of 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% of glycerol, 5% of GTCC, 2.5% of tween-20, 2.5% of polyglycerol-10 oleate, 5% of isononyl isononanoate and 1% of soybean lecithin, dissolving the components by magnetic stirring at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 2% carnosine and 3% VC ethyl ether, 1% nicotinamide and 32% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. Preparing 2mg/mL 2-hydroxypropyl trimethyl ammonium chloride chitosan aqueous solution according to the following steps: whitening co-delivery nanoethosome solution ═ 1:3(w/w) performing ion crosslinking, dripping the whitening co-delivery nano ethosome solution into a 2-hydroxypropyl trimethyl ammonium chloride chitosan solution by using a peristaltic pump (the speed of 30 rpm/min), and stirring at the magnetic stirring speed of 200rpm/min for 30min to obtain the 2-hydroxypropyl trimethyl ammonium chloride chitosan modified whitening co-delivery nano composition liposome; the particle size of the co-delivery nano liposome modified by 2-hydroxypropyl trimethyl ammonium chloride chitosan is detected, and the co-delivery nano liposome is 89.0nm in particle size and 32.4mV in Zeta potential.

Example 4

The preparation method comprises the following steps of carrying out ultrasonic treatment on 5% of phenethyl resorcinol, 1% of glabridin, 10% of 1, 2-propylene glycol and 10% of 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% of glycerol, 5% of GTCC, 2.5% of tween-20, 2.5% of polyglycerol-10 oleate, 5% of isononyl isononanoate and 1% of soybean lecithin, dissolving the components by magnetic stirring at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 2% carnosine and 3% VC ethyl ether, 1% nicotinamide and 32% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. Preparing 4mg/mL 2-hydroxypropyl trimethyl ammonium chloride chitosan aqueous solution according to the following steps: whitening co-delivery nanoethosome solution ═ 1:3(w/w) performing ion crosslinking, dripping the whitening co-delivery nano ethosome solution into a 2-hydroxypropyl trimethyl ammonium chloride chitosan solution by using a peristaltic pump (the speed of 30 rpm/min), and stirring at the magnetic stirring speed of 200rpm/min for 30min to obtain the 2-hydroxypropyl trimethyl ammonium chloride chitosan modified whitening co-delivery nano composition liposome; the particle size of the co-delivery nano liposome modified by 2-hydroxypropyl trimethyl ammonium chloride chitosan is detected, and the co-delivery nano liposome is 96.5nm in particle size and 36.5mV in Zeta potential.

Example 5

The preparation method comprises the following steps of carrying out ultrasonic treatment on 5% of phenethyl resorcinol, 1% of glabridin, 10% of 1, 2-propylene glycol and 10% of 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% of glycerol, 5% of GTCC, 2.5% of tween-20, 2.5% of polyglycerol-10 oleate, 5% of isononyl isononanoate and 1% of soybean lecithin, dissolving the components by magnetic stirring at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 2% carnosine and 3% VC ethyl ether, 1% nicotinamide and 32% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. Preparing 16mg/mL 2-hydroxypropyl trimethyl ammonium chloride chitosan aqueous solution according to the following steps: whitening co-delivery nanoethosome solution ═ 1:3(w/w) performing ion crosslinking, dripping the whitening co-delivery nano ethosome solution into a 2-hydroxypropyl trimethyl ammonium chloride chitosan solution by using a peristaltic pump (the speed of 30 rpm/min), and stirring at the magnetic stirring speed of 200rpm/min for 30min to obtain the 2-hydroxypropyl trimethyl ammonium chloride chitosan modified whitening co-delivery nano composition liposome; the particle size of the 2-hydroxypropyl trimethyl ammonium chloride chitosan modified whitening co-delivery nano liposome is detected, and the co-delivery nano liposome is obtained to be 100.5nm in particle size and 37.6mV in Zeta potential.

Example 6

The preparation method comprises the following steps of carrying out ultrasonic treatment on 5% of phenethyl resorcinol, 1% of glabridin, 10% of 1, 2-propylene glycol and 10% of 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% of glycerol, 1% of GTCC, 2.5% of tween-20, 2.5% of polyglycerol-10 oleate, 5% of isononyl isononanoate and 1% of soybean lecithin, dissolving the components by magnetic stirring at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 1% carnosine and 1% VC ethyl ether, 1% nicotinamide and 39% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. Preparing 8mg/mL 2-hydroxypropyl trimethyl ammonium chloride chitosan aqueous solution according to the following steps: whitening co-delivery nanoethosome solution ═ 1:3(w/w) performing ion crosslinking, dripping the whitening co-delivery nano ethosome solution into a 2-hydroxypropyl trimethyl ammonium chloride chitosan solution by using a peristaltic pump (the speed of 30 rpm/min), and stirring at the magnetic stirring speed of 200rpm/min for 30min to obtain the 2-hydroxypropyl trimethyl ammonium chloride chitosan modified whitening co-delivery nano composition liposome; the particle size of the co-delivery nano liposome modified by 2-hydroxypropyl trimethyl ammonium chloride chitosan is detected, and the co-delivery nano liposome is obtained to be 98.9nm in particle size and 37.5mV in Zeta potential.

Example 7

Performing ultrasonic treatment on 10% phenethyl resorcinol, 1% glabridin, 10% 1, 2-propylene glycol and 10% 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% glycerol, 3% GTCC, 2.5% Tween-20, 2.5% polyglycerol-10 oleate, 5% isononanoic acid isononyl ester and 3% soybean lecithin, stirring and dissolving under magnetic force at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 2% carnosine and 3% VC ethyl ether, 1% nicotinamide and 27% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. Preparing 8mg/mL 2-hydroxypropyl trimethyl ammonium chloride chitosan aqueous solution according to the following steps: whitening co-delivery nanoethosome solution ═ 1:3(w/w) performing ion crosslinking, dripping the whitening co-delivery nano ethosome solution into a 2-hydroxypropyl trimethyl ammonium chloride chitosan solution by using a peristaltic pump (the speed of 30 rpm/min), and stirring at the magnetic stirring speed of 200rpm/min for 30min to obtain the 2-hydroxypropyl trimethyl ammonium chloride chitosan modified whitening co-delivery nano composition liposome; the particle size of the co-delivery nano liposome modified by 2-hydroxypropyl trimethyl ammonium chloride chitosan is detected, and the co-delivery nano liposome is 97.6nm in particle size and 38.0mV in Zeta potential.

Comparative example 1

The preparation method comprises the following steps of carrying out ultrasonic treatment on 5% of phenethyl resorcinol, 1% of glabridin, 10% of 1, 2-propylene glycol and 10% of 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% of glycerol, 5% of GTCC, 2.5% of tween-20, 2.5% of polyglycerol-10 oleate, 5% of isononyl isononanoate and 1% of soybean lecithin, dissolving the components by magnetic stirring at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 2% carnosine and 3% VC ethyl ether, 1% nicotinamide and 32% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. A chitosan solution of 8mg/mL (dissolved in 0.2% acetic acid) was prepared, following the chitosan solution: whitening co-delivery nanoethosome solution ═ 1:3(w/w) carrying out ion crosslinking, dripping the whitening co-delivery nano ethosome solution into a chitosan solution by using a peristaltic pump (the speed of 30 rpm/min), and carrying out magnetic stirring at the speed of 200rpm/min for 30min to obtain a chitosan modified whitening co-delivery nano composition liposome; the particle size of the chitosan modified whitening co-delivery nano liposome is detected, and the co-delivery nano liposome is obtained to be 101.9nm in particle size and 22.9mV in Zeta potential.

Comparative example 2

The preparation method comprises the following steps of carrying out ultrasonic treatment on 5% of phenethyl resorcinol, 1% of glabridin, 10% of 1, 2-propylene glycol and 10% of 1, 3-butanediol to fully dissolve the phenethyl resorcinol and the glabridin, then adding 20% of glycerol, 5% of GTCC, 2.5% of tween-20, 2.5% of polyglycerol-10 oleate, 5% of isononyl isononanoate and 1% of soybean lecithin, dissolving the components by magnetic stirring at normal temperature, and obtaining a phase A for later use after the soybean lecithin is fully dissolved. Weighing 2% carnosine and 3% VC ethyl ether, 1% nicotinamide and 32% deionized water, and dissolving in water to obtain phase B for later use. Adding the phase B into the phase A at a constant speed under magnetic stirring, reacting for 20min, and shearing at a high speed of 8000rpm for 5min to obtain crude liposome. And (3) homogenizing the crude ethosome for 5 times under high pressure and 1.5MPa respectively to obtain the whitening co-delivery nano ethosome. The particle size of the anionic whitening co-delivery nano liposome is detected to be 94.3nm, and the Zeta potential is-40.6 mV.

The samples prepared in example 1, example 2, example 3, example 4, comparative example 1 and comparative example 2 were subjected to a particle size test and a heat and cold resistance stability test.

The test method comprises the following steps:

(1) the test was carried out using a malvern Nano ZS90 particle size potential detector, and the test sample was diluted 500 times with ultrapure water at a test angle of 90 ° and a test temperature of 25 ℃, and repeated 3 times.

(2) Samples of examples 1 to 4, comparative example 1 and comparative example 2 were placed under conditions of heat resistance of 45 ℃, cold resistance of 4 ℃ and temperature of 18 ℃ after completion of preparation, and the particle diameter, PDI, appearance state and odor thereof were measured on the day of preparation (indicated by 0), after one month, after two months and after three months, respectively. The test results are shown in tables 1-6 below:

table 1 stability test results for example 1 sample

Table 2 stability test results for example 2 sample

Table 3 stability test results for example 3 samples

Table 4 stability test results for example 4 sample

Table 5 stability test results for comparative example 1 sample

Table 6 stability test results for comparative example 2 sample

As can be seen from the above tables 1 to 6, the heat resistance, cold resistance stability (particle size, dispersion index (PDI), appearance state and odor) of the samples of examples 1 to 4 for three months were better than those of comparative example 2. Therefore, the grafted chitosan derivative can play a better role in protecting and shielding a whitening composite co-delivery nano liposome system under the action of combining high-pressure microjet, and has a good stability effect on indexes such as particle size, dispersion index (PDI), appearance and smell. In addition, the stability ratio of heat resistance and cold resistance of the samples of examples 1-4 for three months is superior to that of comparative example 1, which shows that the grafted chitosan derivative can play a better protective barrier role for the whitening composite co-delivery nano liposome system compared with natural chitosan.

Whitening efficacy test

The evaluation of the whitening effect of the human body is carried out according to a second method 'a method for testing the whitening effect of removing the freckles by using the freckles for the open human body' in the cosmetic testing method. The test samples were added with 5% of the emulsion of example 1 and 5% of example 2 (indicated by "example 1 group, example 2 group"), 5% of the emulsion of comparative example 1 and 5% of comparative example 2 (indicated by "comparative example 1 group, comparative example 2 group"), and 5% of the emulsion base (indicated by "blank group"), respectively. The test formulations are shown in table 7 below.

Table 7 whitening efficacy test formula table

The preparation process comprises the following steps:

(1) mixing the phase A components, heating to 75-85 ℃, and uniformly stirring for later use;

(2) mixing the components of the phase B, heating to 75-85 ℃ until the components are melted, and uniformly stirring for later use;

(3) adding the phase B into the phase A, and homogenizing at 10000-15000 rpm for 3-5 min;

(4) cooling to 50-60 ℃, adding the component C, continuing emulsifying and homogenizing, and homogenizing at 10000-15000 rpm for 3-5 min;

(5) cooling to 45-55 ℃, adding the component D and the component E, and uniformly stirring;

the test method comprises the following steps: at least 150 eligible, healthy subjects aged 18-60 years were selected and randomly assigned equally to five trial groups, ensuring a 30-unit number of effective cases per group and comparable male to female ratios. The subjects used the respective test samples (example 1, example 2, comparative example 1, comparative example 2 or blank) as required for 8 weeks each after cleansing in the morning and evening, according to the directions of the experimenters and instructions. Other products which claim to be whitening and freckle-removing can not be used in the period. Skin color ITA ° values, skin melanin MI values were tested before, 2 weeks, 4 weeks, 8 weeks after use.

Using an instrument: skin color test probe: colorimeter CL400, manufactured by CK (Courage + Khazaka) of Germany; skin pigment test probe, Mexameter MX18, manufactured by CK (Courage + Khazaka) of Germany.

The larger the skin color ITA DEG value, the whiter the skin color. As shown in fig. 1, volunteers showed a significant increase in skin color ITA ° at week 4 after using the comparative example 1 group, and the increase in skin color ITA ° value at week 8 was 7.98%; the volunteers, after using the group of comparative example 2, had a 4.94% increase in the skin volunteer color ITA ° value at week 8 of use; after the volunteers used the example 1 group and the example 2 group, the skin color ITA DEG value is obviously improved from week 2, the improvement rate of the skin color ITA DEG value at week 8 is 10.53 percent and 9.33 percent respectively, and the effect of improving the skin color is obviously better than that of the comparative example 1 group (7.98 percent) and the comparative example 2 group (4.94 percent).

The smaller the MI value of skin melanin, the lower the melanin content of the skin, and the whiter the skin. Referring to fig. 2, the skin pigment MI values of the example 1 group, the example 2 group, the comparative example 1 group and the comparative example 2 group all showed a decreasing trend with the duration of use, indicating that the skin melanin content was decreased in the example 1 group, the example 2 group, the comparative example 1 group and the comparative example 2 group. Wherein, the reduction rates of the skin melanin MI values at week 2 of the example 1 group and the example 2 group were 8.60% and 6.56%, respectively; the reduction in skin melanin MI values at week 8 was 13.34% and 11.46%, respectively. Whereas the reduction rates of the skin melanin MI values at week 8 of the comparative example 1 group and the comparative example 2 group were 9.80% and 5.79%, respectively, indicating that the examples 1 group and the example 2 group have better effects of reducing the skin melanin MI values than the comparative example 1 group and the comparative example 2 group for the same use time.

The skin color ITA DEG value and skin melanin MI value of the blank group have no significant change trend after 8 weeks of continuous use of the emulsion matrix.

In conclusion, the whitening effect of the groups of example 1 and example 2 is superior to that of the groups of comparative example 1 and comparative example 2, that is, the whitening effect of the cationic whitening co-delivery nano-composition liposome modified by the grafted chitosan derivative prepared by the method of the invention is obviously superior to that of the traditional anionic whitening co-delivery nano-composition liposome; is also superior to the whitening co-delivery nano composition liposome modified by natural chitosan.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims (8)

1. A preparation method of a grafted chitosan derivative modified whitening co-delivery nano composition is characterized by comprising the following steps:

(1) weighing 5-10% of phenethyl resorcinol, 1-5% of glabridin, 5-10% of 1, 2-propylene glycol and 5-10% of 1, 3-butanediol according to the mass percentage, and fully dissolving the phenethyl resorcinol and the glabridin by ultrasonic; then sequentially adding 10-20% of glycerol, 1-10% of GTCC, 1-10% of Tween-20, 1-10% of isononyl isononanoate, 1-5% of soybean lecithin and 1-5% of polyglycerol-10 oleate, and stirring at 25-30 ℃ to fully dissolve all the components to obtain an A phase;

(2) weighing 1-5% of carnosine, 1-5% of Vc ethyl ether, 1-5% of nicotinamide and 20-60% of deionized water in percentage by mass, and performing ultrasonic treatment to completely dissolve all the components to obtain a phase B;

(3) adding the phase B into the phase A at a constant speed under magnetic stirring, stirring at 25-30 deg.C to make the two phases uniformly mixed, continuing to react for 20min, and performing high-speed shearing dispersion by using high-shear homogenizing emulsifying machine to obtain crude ethosome;

(4) carrying out high-pressure homogenization on the crude alcohol body by a high-pressure micro-jet homogenization method to obtain a whitening composite co-delivery nanometer alcohol body;

(5) and (3) dissolving the grafted chitosan derivative in the aqueous solution in advance to obtain a grafted chitosan derivative solution, dripping the whitening composite co-delivery nano ethosome solution obtained in the step (4) into the grafted chitosan derivative solution at a constant speed by using a peristaltic pump under the stirring condition, and continuing stirring for 10-30min after finishing dripping to obtain the grafted chitosan derivative modified whitening co-delivery nano composition liposome.

2. The method for preparing the grafted chitosan derivative modified whitening co-delivery nano-composition according to claim 1, wherein the grafted chitosan derivative modified whitening co-delivery nano-composition comprises the following steps:

the stirring speed in the step (3) is 150-400 rpm/min; the power of the high-shear homogenizing and emulsifying machine in the step (3) is 500-.

3. The method for preparing the grafted chitosan derivative modified whitening co-delivery nano-composition according to claim 1, wherein the grafted chitosan derivative modified whitening co-delivery nano-composition comprises the following steps:

the high-pressure homogenizing pressure in the step (4) is 1.0-1.5MPa, and the circulation is performed for 3-6 times.

4. The method for preparing the grafted chitosan derivative modified whitening co-delivery nano-composition according to claim 1, wherein the grafted chitosan derivative modified whitening co-delivery nano-composition comprises the following steps:

the grafted chitosan derivative in the step (5) is at least one of N-alkyl chitosan, O-alkyl chitosan, lauryl-succinylated chitosan, carboxymethyl chitosan, 2-hydroxypropyl trimethyl ammonium chloride chitosan, chitosan trimethyl quaternary ammonium salt, polyacrylamide grafted chitosan, polylactic acid grafted chitosan, polycaprolactone grafted chitosan and polyvinyl alcohol grafted chitosan.

5. The method for preparing the grafted chitosan derivative modified whitening co-delivery nano-composition according to claim 1, wherein the grafted chitosan derivative modified whitening co-delivery nano-composition comprises the following steps:

the concentration of the grafted chitosan derivative in the grafted chitosan derivative solution in the step (5) is 2-16 mg/mL;

the mass ratio of the grafted chitosan derivative solution to the whitening composite co-delivery nanometer ethosome solution in the step (5) is 1: 2-1: 5.

6. the method for preparing the grafted chitosan derivative modified whitening co-delivery nano-composition according to claim 1, wherein the grafted chitosan derivative modified whitening co-delivery nano-composition comprises the following steps:

the stirring speed in the step (5) is 150-400rpm/min, and the dropping speed of the peristaltic pump is 20-50 rpm/min.

7. A grafted chitosan derivative modified whitening co-delivery nano-composition prepared by the method of any one of claims 1 to 6.

8. The use of the grafted chitosan derivative modified whitening co-delivery nano-composition of claim 7 in cosmetics.

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