CN114917134B

CN114917134B - Freeze-dried composition containing skeleton molecules and liposome embedding and application thereof

Info

Publication number: CN114917134B
Application number: CN202210636879.3A
Authority: CN
Inventors: 吴冬; 候丽丽; 梁乾宝
Original assignee: Meishang Guangzhou Cosmetics Co ltd
Current assignee: Wu Dong
Priority date: 2022-06-07
Filing date: 2022-06-07
Publication date: 2023-08-15
Anticipated expiration: 2042-06-07
Also published as: CN114917134A

Abstract

The invention discloses a freeze-dried composition containing skeleton molecules and liposome embedding and application thereof, and relates to the technical field of freeze-dried compositions. The binary freeze-dried composition comprises the following components in percentage by weight: 55-80% of skeleton molecules and 20-45% of liposome embedding. The invention mutually coordinates the framework molecules and liposome embedding, and the formed freeze-dried preparation can freeze-dried oil and fat, polyalcohol, easily decomposed unstable active substances, easily formed saccharides after drying and other components which are difficult to freeze-dry conventionally, and has the advantages of high effective component load, good stability, high compatibility of oily and aqueous components and good skin feel.

Description

Freeze-dried composition containing skeleton molecules and liposome embedding and application thereof

Technical Field

The invention relates to the technical field of freeze-dried compositions, in particular to a freeze-dried composition containing skeleton molecules and liposome embedding and application thereof.

Background

The facial skin care product is a cosmetic for the face, and contains precious functional components such as plant extracts and the like, so that the skin care product has the effects of preventing aging, resisting wrinkle, preserving moisture, whitening, removing freckles and the like. The traditional facial skin care products usually contain unstable active substances, so that the long-term storage of the skin care products is affected, and the active substances are required to be better stored and delivered due to the lipophilic property of skin, so that a large amount of emulsifying agents, skin feel regulators and the like are required to be added for improving the stability of the products, the traditional facial skin care products are in bottled liquid form, preservatives are required to be added for preventing bacteria from breeding, the problem of inconvenience in carrying exists, and the skin is damaged due to the risk of cross contamination caused by multiple switching, so that the cosmetic ultra-low temperature vacuum freeze drying technology is developed.

Many frozen cosmetics such as freeze-dried masks and cosmetic freeze-dried powders, etc. are now on the market, and they are all produced by freeze-drying technology. The freeze drying technology is to freeze the matter with great amount of water to form solid, and sublimate the solid water directly under vacuum condition while maintaining the matter inside the frozen ice shelf to preserve the active components of biological matter fully.

A polypeptide lyophilized powder mask composition and methods of making and using the same are disclosed in patent CN111632005a, for example. The polypeptide freeze-dried powder mask composition consists of polypeptide freeze-dried powder, a freeze-dried solvent, mask liquid and mask cloth. The freeze-drying technology is adopted to store the materials such as oligopeptide-5, oligopeptide-1 and the like in a freeze-dried powder state, a layer of protective film is formed on the surface of the material to be freeze-dried by adding trehalose and mannitol, the freeze-drying process of the material is effectively protected, meanwhile, a good freeze-drying skeleton is provided for the material in the freeze-drying process, and the material is shaped. Said invention not only retains the bioactive substances and heat-sensitive substances in the materials of oligopeptide-5 and oligopeptide-1, but also optimizes the storage condition of the materials of oligopeptide-5 and oligopeptide-1, and possesses the advantages of high active component and maximally retaining the bioactive substances and heat-sensitive substances in the materials, but the preservative is added into the above-mentioned polypeptide freeze-dried powder surface film, so that it possesses a certain irritation to skin, and its safety is reduced.

Also disclosed in patent CN112315821a is a hydrolyzed protein liposome and a preparation method thereof. The hydrolyzed protein liposome consists of a cosmetic active ingredient, a liposome framework ingredient, a guest ingredient and a freeze-drying protective agent. The liposome is used as a carrier for hydrolyzing keratin, lactose, mannitol and trehalose are used as freeze-drying protective agents, so that on one hand, the transdermal absorption of the hydrolyzed keratin is promoted, and on the other hand, the concentration of active substances is continuously increased in the process of contacting with skin. However, the freeze-dried facial mask system only contains water-based substances, and the oil components are not freeze-dried, so that the obtained facial mask has insufficient skin feel and low experience.

Therefore, there is an urgent need for a lyophilized preparation which is free of preservatives and chemical additives, has a low weight without water, and can be used for lyophilizing oil components.

Disclosure of Invention

Aiming at the defects of the prior art, the application aims to provide a freeze-dried preparation which has high effective component load, high compatibility of oily components and aqueous components, multiple dimensions of skin feel, good experience feel, low moisture content and light weight, and does not need to add chemical synthetic oil, skin feel regulator, preservative and the like.

In order to achieve the above purpose, the present application adopts the following technical scheme:

The first object of the invention is to provide a freeze-dried composition containing a framework molecule and liposome embedding, which comprises the following components in percentage by weight: 55-80% of skeleton molecules and 20-45% of liposome embedding.

Preferably, the freeze-dried composition comprises the following components in percentage by weight: 60-75% of framework molecules and 25-45% of liposome embedding.

Still preferably, the freeze-dried composition comprises the following components in percentage by weight: 75% of backbone molecules and 25% of liposomes.

Wherein the backbone molecule comprises an amino acid;

the amino acid is one or more of proline, tryptophan, sodium glutamate, alanine, glycine, lysine hydrochloride, sarcosine, L-tyrosine, phenylalanine and arginine;

preferably, the amino acid is glycine.

The amino acid has acid and alkali, so that the pH of the solution can be kept stable in the low-temperature preservation and freeze drying processes of biological products, thereby achieving the purpose of protecting active components; the crystallization glycine can raise the collapse temperature of the finished product and prevent the damage of protein medicines caused by collapse.

The skeleton molecule also comprises two or more than two of trehalose, hydrolyzed xylan, mannitol and rhamnose;

the trehalose is a dimer of glucose, is stable non-reducing disaccharide, is widely used for freeze-drying preservation, has relatively stable biological performance as a sample freeze-dried as a protective agent, has long preservation time, is an optimal protective agent in the low-temperature biological field, has smaller molecular weight, is easy to fill in gaps in macromolecules, effectively limits the structural change in the macromolecules, avoids the inactivation of active substances, has high glass transition temperature, and can effectively prevent disintegration and collapse. Mannitol is a polyhydroxy compound which can form a loose and firm uniform framework as a carrier, can also be used as a permeability regulator and a freeze-drying protective agent for proteins, is generally used as a filling agent in the freeze-drying process of biological products, and can be crystallized during slow freezing, so that a supporting structure is provided for an active component, and meanwhile, mannitol cannot react with the active component. Hydrolyzed xylan, rhamnose and other sugars are the most common and most widely used class of lyoprotectants, which protect the active during the various stages of lyophilization.

Preferably, the backbone molecule is a mixture of glycine, trehalose, hydrolyzed xylan, mannitol, and rhamnose;

the mass ratio of glycine to trehalose to hydrolyzed xylan to mannitol to rhamnose is 1:1:1:1.

The liposome is embedded in lecithin; preferably, the lecithin is soybean lecithin and/or hydrogenated lecithin; still more preferably, the lecithin is a mixture of soybean lecithin and hydrogenated lecithin; still preferably, the mass ratio of the soybean lecithin to the hydrogenated lecithin is 1:3.

The choline group of the lecithin is a lipophilic group and has the function of dispersing one phase of two mutually-insoluble phases (oil phase and water phase) in the other phase, and the soybean lecithin and the hydrogenated lecithin with the mass ratio of 1:3 are adopted as raw materials for embedding the liposome, so that the oil phase and the water phase can be better dispersed to form stable emulsion.

In the implementation process, the mass ratio of skeleton molecules to liposome embedding is reasonably controlled, and particularly the mass ratio of glycine, trehalose, hydrolyzed xylan, mannitol and rhamnose and the mass ratio of soybean lecithin and hydrogenated lecithin are controlled, so that the obtained composition can better freeze-dry one or more of grease components, polyalcohol components, unstable and easily-decomposed components, easily-gelling components after drying, vitamins, thickening agents and active ingredients.

Therefore, another object of the present application is to provide a cosmetic lyophilized preparation, comprising the above lyophilized composition, and further comprising one or more of a lipid component, a polyol component, an unstable and easily decomposed component, a easily gelling component after drying, a vitamin, a thickener and an active ingredient;

the grease component is selected from animal-derived grease and plant-derived grease; the animal-derived oil is one or more of butter, mink oil, egg yolk oil, lanolin oil and squalane; the vegetable oil is one or more selected from shea butter, hawaii fruit oil, jojoba seed oil, olive oil, coconut oil, castor oil, cotton seed oil, soybean oil, sesame oil, almond oil, peanut oil, corn oil, rice bran oil, tea seed oil, sea buckthorn oil, avocado oil, chestnut oil, walnut oil and cocoa butter; the polyalcohol component is selected from glycerol or/and propylene glycol; the unstable and easily-decomposed component is selected from retinol palmitate and/or hydroxy pinacolone retinol sulfonate; the components easy to form gel after drying are selected from glass color factors.

The vitamin is vitamin C and its salt; the thickening agent is one or more selected from microbial gum, xanthan gum and sclerotium gum; the active ingredients are one or more of adenosine, tocopheryl acetate, superoxide dismutase, sodium hyaluronate, anthocyanin, hydrolyzed ginsenoside, centella asiatica extract and ganoderma lucidum extract.

A third object of the present invention is to provide a method for preparing the above-mentioned cosmetic lyophilized preparation, comprising the steps of:

(1) Weighing skeleton molecules, vitamins, thickening agents and active ingredients in the formula dosage, uniformly mixing and sieving to obtain a mixture A;

(2) Adding water into liposome embedding with the formula dosage, stirring and dissolving at 45 ℃, adding the mixture A in the step (1), stirring and emulsifying and embedding at 45 ℃ for 30min to obtain a mixture B;

(3) Loading one or more of a grease component, a polyol component, an unstable and easily-decomposed component and an easily-gelled component after drying into the mixture B obtained in the step (2), and performing ultrasonic emulsification to obtain a mixture C; then shaping the mixture C to obtain a shaped mixture C;

(4) Carrying out freeze-drying treatment on the molded mixture C obtained in the step (3) to obtain the freeze-dried preparation;

the freeze-drying comprises three steps of pre-freezing, sublimation, analysis and drying;

the pre-freezing operation is as follows: starting the freeze-drying equipment to enable the temperature of the cold well to reach below minus 45 ℃ from the room temperature for about 40 minutes, and keeping for 2 hours;

the sublimation is divided into three stages, and the specific operation is as follows:

stage one: slowly heating the temperature in the freeze-drying bin to about minus 30 ℃ within 1 hour, and keeping the vacuum degree below 1 Pa for 3-5 hours;

Step two, slowly heating the temperature in the freeze-drying bin to about minus 10 ℃ within 1 hour, and keeping the vacuum degree at 25-35 Pa for 4-8 hours;

and step three, slowly raising the temperature to about 0 ℃ in a freeze-drying bin within 1 hour, and keeping the vacuum degree at 35-45 Pa for 1-2 hours.

The analytical drying is divided into two stages, and the specific operation is as follows:

stage one: the temperature in the freeze-drying bin reaches 5-15 ℃ within 1 hour, the vacuum degree is 45-55 Pa, and the temperature is kept for 2-3 hours;

stage two: the temperature in the freeze drying bin reaches 15-25 ℃ within 1 hour, the vacuum degree is reduced to 35-45 Pa, and the temperature is kept for 1-2 hours.

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

firstly, the components which are difficult to freeze-dry in the conventional way, such as high-loading and high-compatibility freeze-dry oil, polyalcohol, easily decomposed unstable active substances, easily formed glue after drying, and the like;

secondly, the skin feel is good, no chemical components, preservatives and the like are added, and the shaping and the solubility are good;

thirdly, the water-free weight is lighter, and the carrying and long-distance transportation are convenient.

Drawings

FIG. 1 is a view showing the state of a knife-cut surface and the state of reconstitution of a lyophilized preparation prepared in application examples 1.1 to 1.8;

FIG. 2 is a graph showing the effect of the lyophilized preparation prepared in application examples 1.1 to 1.8 after being left at 40℃for 2 weeks at room temperature;

FIG. 3 is a view showing the state of a knife-cut surface and the state of reconstitution of the lyophilized preparation prepared in application examples 2.1 to 2.8;

FIG. 4 is a graph showing the effect of the lyophilized preparation prepared in application examples 2.1-2.8 after being left at 40℃for 2 weeks at room temperature;

FIG. 5 is a view showing the state of a knife-cut surface and the state of reconstitution of the lyophilized preparation prepared in application examples 3.1 to 3.8;

FIG. 6 is a graph showing the effect of the lyophilized preparation prepared in application examples 3.1 to 3.8 after being left at 40℃for 2 weeks at room temperature;

FIG. 7 is a view showing the state of a knife-cut surface and the state of reconstitution of the lyophilized preparation prepared in application examples 4.1 to 4.8;

FIG. 8 is a graph showing the effect of the lyophilized preparation prepared in application examples 4.1 to 4.8 after being left at 40℃for 2 weeks at room temperature;

FIG. 9 is a view showing the state of a knife-cut surface and the state of reconstitution of the lyophilized preparation prepared in application examples 5.1 to 5.8;

FIG. 10 is a graph showing the effect of the lyophilized preparation prepared in application examples 5.1 to 5.8 after being left at 40℃for 2 weeks at room temperature;

FIG. 11 is a view showing the state of a knife-cut surface and the state of reconstitution of the lyophilized preparation prepared in application examples 6.1 to 6.8;

FIG. 12 is a graph showing the effect of the lyophilized formulations prepared in application examples 6.1 to 6.8 after being left at 40℃for 2 weeks at room temperature;

FIG. 13 is a graph showing a comparison of the maximum load amounts of the respective components in the lyophilized formulations prepared in application examples 1 to 6;

FIG. 14 is a HPLC plot of Hydroxy Pinacolone Retinoate (HPR) before and after lyophilization of the lyophilized formulation prepared in application example 1.7;

wherein A is HPR standard substance HPLC spectrum, B is HPLC spectrum before freeze-drying of the preparation, and C is HPLC spectrum of freeze-dried preparation;

FIG. 15 is an HPLC chart showing retinol palmitate content after freeze-drying and after 2 weeks of high temperature treatment at 40℃after freeze-drying of the lyophilized preparation prepared in application example 1.6;

wherein A is the HPLC profile of retinol palmitate standard substance, B is the HPLC profile of freeze-dried preparation, and C is the HPLC profile of freeze-dried preparation after being treated at 40 ℃ for 2 weeks;

fig. 16 is a state diagram after reconstitution of the lyophilized mask formulation prepared in application example 7;

FIG. 17 is a dissolution profile of the formulation of comparative example 1 and a lyophilized formulation prepared therefrom;

FIG. 18 is a dissolution profile of the formulation of comparative example 2;

fig. 19 is a dissolution profile of the formulation of comparative example 3.

Detailed Description

The above-mentioned features of the invention, or of the embodiments, may be combined in any desired manner. All of the features explained in this specification can be used in combination with any form of method, and each feature disclosed in this specification can be replaced by any alternative feature serving the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, the disclosed features are merely general examples of equivalent or similar features.

The application will be further illustrated with reference to specific examples. These examples are only for illustrating the present application and are not intended to limit the scope of the present application. The following examples are presented to illustrate specific conditions, generally according to conventional conditions or according to manufacturer's recommended conditions. All percentages and fractions are by weight unless specifically indicated.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present application. The preferred embodiments and materials described herein are exemplary only.

The raw materials used in the experimental process and the purchase manufacturer and model of the instrument are as follows:

examples 1-6A lyophilized composition comprising a backbone molecule and liposome entrapment

The weight percentage of the components is as follows:

application examples 1.1-1.8 a freeze-dried preparation and preparation method thereof

The composite material comprises the following components in parts by weight:

the method comprises the following steps:

(1) Weighing glycine, xanthan gum and anthocyanin which are used in the formula, uniformly mixing, and sieving with a 150-mesh sieve to obtain a mixture A;

(2) Adding water into soybean lecithin with the formula dosage, stirring and dissolving at 45 ℃, adding the mixture A in the step (1), stirring, emulsifying and embedding at 45 ℃ for 30min to obtain a mixture B;

(3) Heating and melting animal source butter, adding into the mixture B in the step (2), and emulsifying by ultrasonic (at 40 ℃ C., 300W,30 min) to obtain a mixture C; then shaping the mixture C to obtain a shaped mixture C;

(4) Placing the molded mixture C obtained in the step (3) in a low-temperature environment for freeze-drying treatment to obtain the cosmetic freeze-dried preparation;

the freeze-drying treatment comprises three steps of pre-freezing, sublimation, analysis and drying;

The preparation methods described in application examples 1.2 to 1.8 were the same as those of application example 1.1 except that the components added in step (3) were different.

Detection result:

as shown in figure 1, freeze-dried samples were prepared according to application examples 1.1-1.8, and the obtained freeze-dried balls were well-formed, complete in morphology and smooth in surface; the knife cut surface layer is in a lamination shape; the emulsion is quick in rehydration, can be dissolved in 2 minutes and has good emulsification effect.

Test example 1-1 detection of the emulsion value OD500

According to the emulsion stability evaluation and detection method of protein emulsion, namely the emulsion activity index, the absorbance value of a sample at 500nm is detected, and the emulsion and stability of the sample can be evaluated in a correlated manner; the lyophilized formulations prepared in application examples 1.1 to 1.8 and the lyophilized and reconstituted water to examine the OD500 value thereof were as shown in Table 1 below.

TABLE 1

	OD500 value of the formulation	OD500 value of rehydration after lyophilization	Change value
				Application example 1.1	2.603	2.708	4.03％
Application example 1.2	2.426	2.417	-0.37％
				Application example 1.3	2.623	2.633	0.38％
Application example 1.4	2.331	2.306	-1.07％
				Application example 1.5	2.337	2.284	-2.27％
Application example 1.6	2.572	2.668	3.73％
				Application example 1.7	2.667	2.715	1.80％
Application example 1.8	2.563	2.538	-0.98％

Conclusion: the freeze-dried preparation prepared in application examples 1.1-1.8 and the OD500 value of re-dissolved in water after freeze drying have a variation range within +/-5%, which shows that the emulsion effect after re-hydration is good and the emulsion stability is good.

Test examples 1-2 high temperature stability test

Referring to fig. 2, the lyophilized formulations prepared in examples 1.1 to 1.8 were applied, and each of the 3 bottles was subjected to a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and was taken out after 2 weeks, and compared with the sample after 2 weeks at room temperature, the color morphology was unchanged, indicating high temperature stability.

Test examples 1-3 maximum load detection

The determination criteria for the maximum load amount are: the freeze-dried balls are well molded, and the surface is smooth and free of liquid substances; the emulsion is obtained after rehydration, and has no oil drop and no layering.

The maximum loading of the lyophilized composition described in example 1 was tested for 10% of the maximum loading of the lyophilized composition described in example 1 for animal-derived butter; the maximum load for shea butter is 1%; the maximum load for hawaii fruit oil is 10%; the maximum load for glycerol is 1%; the maximum load for propylene glycol was 1%; maximum loading for retinol palmitate was 1%; the maximum load for HPR is 1%; the maximum load for the glass factor is 5%.

Application examples 2.1-2.8 a freeze-dried preparation and preparation method thereof

The composite material comprises the following components in parts by weight:

the method comprises the following steps:

(1) Weighing glycine, trehalose, xanthan gum and anthocyanin which are used in the formula, uniformly mixing, and sieving with a 150-mesh sieve to obtain a mixture A;

(2) Adding water into soybean lecithin and hydrogenated lecithin with the formula dosage, stirring and dissolving at 45 ℃, adding the mixture A in the step (1), stirring, emulsifying and embedding at 45 ℃ for 30min to obtain a mixture B;

The preparation methods described in application examples 2.2 to 2.8 were the same as those of application example 2.1 except that the components added in step (3) were different.

Detection result:

as shown in fig. 3, the freeze-dried sample was prepared for the above application examples 2.1 to 2.8, and the obtained freeze-dried pellet was well-formed, complete in morphology and smooth in surface; the knife cut surface layer is in a lamination shape; the emulsion is quick in rehydration, can be dissolved in 2 minutes and has good emulsification effect.

Test example 2-1 detection of the emulsion value OD500

According to the emulsion stability evaluation and detection method of protein emulsion, namely the emulsion activity index, the absorbance value of a sample at 500nm is detected, and the emulsion and stability of the sample can be evaluated in a correlated manner; the lyophilized formulations prepared in application examples 2.1 to 2.8 and the lyophilized and reconstituted water to examine the OD500 value thereof were as shown in Table 2 below.

TABLE 2

	OD500 value of the formulation	OD500 value of rehydration after lyophilization	Change value
				Application example 2.1	2.815	2.878	2.24％
Application example 2.2	2.725	2.717	-0.29％
				Application example 2.3	2.623	2.633	0.38％
Application example 2.4	2.536	2.561	0.99％
				Application example 2.5	2.413	2.446	1.37％
Application example 2.6	2.471	2.588	4.73％
				Application example 2.7	2.567	2.513	-2.10％
Application example 2.8	2.571	2.449	-4.75％

Conclusion: the freeze-dried preparation prepared in application examples 2.1-2.8 and the OD500 value of re-dissolved in water after freeze drying have a variation range within +/-5%, which shows that the emulsion effect after re-hydration is good and the emulsion stability is good.

Test example 2-2 high temperature stability test

Referring to fig. 4, the lyophilized formulations prepared in examples 1.1 to 1.8 were applied, and each of the 3 bottles was subjected to a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and was taken out after 2 weeks, and compared with the sample after 2 weeks at room temperature, the color morphology was unchanged, indicating high temperature stability.

Test examples 2-3 maximum load detection

The maximum loading of the lyophilized composition described in example 2 was measured for 15% of the maximum loading of the lyophilized composition described in example 2 for animal-derived butter; the maximum load for shea butter is 5%; the maximum load for hawaii fruit oil is 15%; the maximum load for glycerol is 3%; the maximum load for propylene glycol is 3%; the maximum load for retinol palmitate was 3%; the maximum load for HPR is 3%; the maximum load for the glass factor is 10%.

Application examples 3.1-3.8 a freeze-dried preparation and preparation method thereof

The composite material comprises the following components in parts by weight:

the preparation method of the freeze-dried preparation described in application example 3.1 comprises the following steps: the method comprises the following steps:

(1) Weighing glycine, trehalose, mannitol, rhamnose, xanthan gum and anthocyanin which are used in the formula, uniformly mixing, and sieving with a 150-mesh sieve to obtain a mixture A;

(2) Adding water into hydrogenated lecithin with the formula dosage, stirring and dissolving at 45 ℃, adding the mixture A in the step (1), stirring, emulsifying and embedding at 45 ℃ for 30min to obtain a mixture B;

The preparation methods described in application examples 3.2 to 3.8 were the same as those of application example 3.1 except that the components added in step (3) were different.

Detection result:

as shown in fig. 5, the freeze-dried sample was prepared for the above application examples 3.1 to 3.8, and the obtained freeze-dried pellet was well-formed, complete in morphology and smooth in surface; the knife cut surface layer is in a lamination shape; the emulsion is quick in rehydration, can be dissolved in 2 minutes and has good emulsification effect.

Test example 3-1 detection of the emulsion value OD500

According to the emulsion stability evaluation and detection method of protein emulsion, namely the emulsion activity index, the absorbance value of a sample at 500nm is detected, and the emulsion and stability of the sample can be evaluated in a correlated manner; the lyophilized formulations prepared in application examples 3.1 to 3.8 and the lyophilized and reconstituted water to examine the OD500 value thereof were as shown in Table 3 below.

TABLE 3 Table 3

	OD500 value of the formulation	OD500 value of rehydration after lyophilization	Change value
				Application example 3.1	2.913	2.859	-1.85％
Application example 3.2	2.673	2.746	2.73％
				Application example 3.3	3.021	2.876	-4.80％
Application example 3.4	2.563	2.638	2.93％
				Application example 3.5	2.531	2.584	2.09％
Application example 3.6	2.678	2.789	4.14％
				Application example 3.7	2.664	2.601	-2.36％
Application example 3.8	2.567	2.532	-1.36％

Conclusion: the freeze-dried preparation prepared in application examples 3.1-3.8 and the OD500 value of re-dissolved in water after freeze drying have a variation range within +/-5%, which shows that the emulsion effect after re-hydration is good and the emulsion stability is good.

Test example 3-2 high temperature stability test

Referring to fig. 6, the lyophilized formulations prepared in examples 1.1 to 1.8 were applied, and each of the 3 bottles was subjected to a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and was taken out after 2 weeks, and compared with the sample after 2 weeks at room temperature, the color morphology was unchanged, indicating high temperature stability.

Test example 3-3 maximum load detection

The maximum loading of the lyophilized composition described in example 3 was 15% for animal source butter, as measured for the highest loading of the lyophilized composition described in example 3; the maximum load for shea butter is 5%; the maximum load for hawaii fruit oil is 15%; the maximum load for glycerol is 3%; the maximum load for propylene glycol is 3%; the maximum load for retinol palmitate was 3%; the maximum load for HPR is 3%; the maximum load for the glass factor is 10%.

Application examples 4.1-4.8 a freeze-dried preparation and preparation method thereof

The composite material comprises the following components in parts by weight:

the preparation method of the freeze-dried preparation described in application example 4.1 comprises the following steps: the method comprises the following steps:

(1) Weighing glycine, trehalose, hydrolyzed xylan, mannitol, xanthan gum and anthocyanin which are used in the formula, uniformly mixing, and sieving with a 150-mesh sieve to obtain a mixture A;

(2) Adding water into the hydrogenated lecithin and the soybean lecithin with the formula dosage, stirring and dissolving at 45 ℃, adding the mixture A in the step (1), stirring, emulsifying and embedding at 45 ℃ for 30min to obtain a mixture B;

The preparation methods described in application examples 4.2 to 4.8 were the same as those of application example 4.1 except that the components added in step (3) were different.

Detection result:

as shown in fig. 7, the freeze-dried sample was prepared for the above application examples 4.1 to 4.8, and the obtained freeze-dried pellet was well-formed, complete in morphology and smooth in surface; the knife cut surface layer is in a lamination shape; the emulsion is quick in rehydration, can be dissolved in 2 minutes and has good emulsification effect.

Test example 4-1 detection of the emulsion value OD500

According to the emulsion stability evaluation and detection method of protein emulsion, namely the emulsion activity index, the absorbance value of a sample at 500nm is detected, and the emulsion and stability of the sample can be evaluated in a correlated manner; the lyophilized formulations prepared in application examples 4.1 to 4.8 and the lyophilized and reconstituted water to examine the OD500 value thereof were as shown in Table 4 below.

TABLE 4 Table 4

	OD500 value of the formulation	OD500 value of rehydration after lyophilization	Change value
				Application example 4.1	3.011	2.906	-3.49％
Application example 4.2	2.636	2.619	-0.64％
				Application example 4.3	2.917	2.906	-0.38％
Application example 4.4	2.532	2.625	3.67％
				Application example 4.5	2.535	2.661	4.97％
Application example 4.6	2.637	2.751	4.32％
				Application example 4.7	2.631	2.717	3.27％
Application example 4.8	2.568	2.443	-4.87％

Conclusion: the freeze-dried preparation prepared in application examples 4.1-4.8 and the OD500 value of re-dissolved in water after freeze drying have a variation range within +/-5%, which shows that the emulsion effect after re-hydration is good and the emulsion stability is good.

Test example 4-2 high temperature stability test

Referring to fig. 8, the lyophilized formulations prepared in examples 1.1 to 1.8 were applied, and each of the 3 bottles was subjected to a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and was taken out after 2 weeks, and compared with the sample after 2 weeks at room temperature, the color morphology was unchanged, indicating high temperature stability.

Test example 4-3 maximum load detection

The highest loading of the lyophilized composition described in example 4 was tested and the maximum loading of the lyophilized composition described in example 4 was 30% for animal source butter; the maximum load for shea butter is 5%; the maximum load for hawaii fruit oil is 30%; the maximum load for glycerol is 5%; the maximum load for propylene glycol is 5%; the maximum load for retinol palmitate was 5%; the maximum load for HPR is 5%; the maximum load for the glass factor is 20%.

Application examples 5.1-5.8 a freeze-dried preparation and preparation method thereof

The composite material comprises the following components in parts by weight:

the preparation method of the freeze-dried preparation described in application example 5.1 comprises the following steps: the method comprises the following steps:

(1) Uniformly mixing glycine, trehalose, hydrolyzed xylan, mannitol, rhamnose, xanthan gum and anthocyanin with the formula dosage, and sieving with a 150-mesh sieve to obtain a mixture A;

The preparation methods described in application examples 5.2 to 5.8 were the same as those of application example 5.1 except that the components added in step (3) were different.

Detection result:

as shown in fig. 9, the freeze-dried sample was prepared for the above application examples 5.1 to 5.8, and the obtained freeze-dried pellet was well-formed, complete in morphology and smooth in surface; the knife cut surface layer is in a lamination shape; the emulsion is quick in rehydration, can be dissolved in 2 minutes and has good emulsification effect.

Test example 5-1 detection of the emulsion value OD500

According to the emulsion stability evaluation and detection method of protein emulsion, namely the emulsion activity index, the absorbance value of a sample at 500nm is detected, and the emulsion and stability of the sample can be evaluated in a correlated manner; the lyophilized formulations prepared in application examples 5.1 to 5.8 and the lyophilized and reconstituted water to examine the OD500 value thereof were as shown in Table 5 below.

TABLE 5

Conclusion: the freeze-dried preparation prepared in application examples 5.1-5.8 and the OD500 value of re-dissolved in water after freeze drying have a variation range within +/-5%, which shows that the emulsion effect after re-hydration is good and the emulsion stability is good.

Test example 5-2 high temperature stability test

Referring to fig. 10, the lyophilized formulations prepared in examples 1.1 to 1.8 were applied, and each of the 3 bottles was subjected to a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and was taken out after 2 weeks, and compared with the sample after 2 weeks at room temperature, the color morphology was unchanged, indicating high temperature stability.

Test example 5-3 maximum load detection

The highest loading of the lyophilized composition described in example 5 was tested and the maximum loading of the lyophilized composition described in example 5 was 50% for animal source butter; the maximum load for shea butter is 10%; the maximum load for hawaii oil is 50%; the maximum load for glycerol is 10%; the maximum load for propylene glycol was 10%; the maximum load for retinol palmitate was 10%; the maximum load for HPR is 10%; the maximum load for the glass factor is 30%.

Application examples 6.1-6.8 a freeze-dried preparation and preparation method thereof

The composite material comprises the following components in parts by weight:

the preparation method of the freeze-dried preparation described in application example 6.1 comprises the following steps: the method comprises the following steps:

(3) Heating and melting animal source butter, adding the melted animal source butter into the mixture B in the step (2), and performing ultrasonic emulsification to obtain a mixture C; then shaping the mixture C to obtain a shaped mixture C;

The preparation methods described in application examples 6.2 to 6.8 were the same as those of application example 6.1 except that the components added in step (3) were different.

Detection result:

as shown in fig. 11, the freeze-dried sample was prepared for the above application examples 6.1 to 6.8, and the obtained freeze-dried pellet was well-formed, complete in morphology and smooth in surface; the knife cut surface layer is in a lamination shape; the emulsion is quick in rehydration, can be dissolved in 2 minutes and has good emulsification effect.

Test example 6-1 detection of the emulsion value OD500

According to the emulsion stability evaluation and detection method of protein emulsion, namely the emulsion activity index, the absorbance value of a sample at 500nm is detected, and the emulsion and stability of the sample can be evaluated in a correlated manner; the lyophilized formulations prepared in application examples 6.1 to 6.8 and the lyophilized and reconstituted water to examine the OD500 value thereof were as shown in Table 6 below.

TABLE 6

	OD500 value of the formulation	OD500 value of rehydration after lyophilization	Change value
				Application example 6.1	3.016	3.106	2.98％
Application example 6.2	2.714	2.689	-0.92％
				Application example 6.3	3.027	2.963	-2.11％
Application example 6.4	2.635	2.689	2.05％
				Application example 6.5	2.623	2.693	2.67％
Application example 6.6	2.652	2.768	4.37％
				Application example 6.7	2.707	2.772	2.40％
Application example 6.8	2.763	2.791	1.01％

Conclusion: the freeze-dried preparation prepared in application examples 6.1-6.8 and the OD500 value of re-dissolved in water after freeze drying have a variation range within +/-5%, which shows that the emulsion effect after re-hydration is good and the emulsion stability is good.

Test example 6-2 high temperature stability test

Referring to fig. 12, the lyophilized formulations prepared in examples 1.1 to 1.8 were applied, and each of the 3 bottles was subjected to a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and was taken out after 2 weeks, and compared with the sample after 2 weeks at room temperature, the color morphology was unchanged, indicating high temperature stability.

Test example 6-3 maximum load detection

The highest loading of the lyophilized composition described in example 6 was tested and the maximum loading of the lyophilized composition described in example 6 was 30% for animal source butter; the maximum load for shea butter is 5%; the maximum load for hawaii fruit oil is 30%; the maximum load for glycerol is 10%; the maximum load for propylene glycol was 10%; the maximum load for retinol palmitate was 5%; the maximum load for HPR is 5%; the maximum load for the glass factor is 30%.

As can be seen from fig. 13, the specific components embedded in the skeleton molecule and the liposome have a larger influence on the loading amounts of the grease component, the polyol component, the unstable and easily-decomposed component and the easily-gelled component after drying, in the embodiment 1, glycine is taken as the skeleton molecule, and soybean lecithin is taken as the liposome for embedding, so that the obtained freeze-dried composition has a smaller loading amount of the grease component, the polyol component, the unstable and easily-decomposed component and the easily-gelled component after drying; in the embodiment 2, glycine and trehalose are taken as skeleton molecules, soybean lecithin and hydrogenated lecithin are taken as liposome for embedding, and the obtained freeze-dried composition has improved loading capacity for grease components, polyalcohol components, unstable components easy to decompose and components easy to gel after drying compared with the embodiment 1, but still has lower loading capacity; in the embodiment 3 and the embodiment 4, a plurality of components are added, and the loading amounts of the grease component, the polyalcohol component, the unstable and easily-decomposed component and the easily-glued component of the obtained freeze-dried composition are obviously improved in the embodiment 1-2; example 5 by controlling the backbone molecules to be glycine, trehalose, hydrolyzed xylan, mannitol and rhamnose in a mass ratio of 1:1:1:1:1; the liposome is embedded into soybean lecithin and hydrogenated lecithin in a mass ratio of 1:3, so that the loading capacity of grease components, polyalcohol components, unstable components easy to decompose and components easy to gel after drying can be obviously improved; the components of the lyophilized composition disclosed in example 6 are the same as in example 5, and the content of the scaffold molecules and liposome entrapment is the same as in example 5, but the loading amounts of the lipid component, the polyol component, the unstable and easily decomposed component and the easily gel-forming component after drying are significantly reduced due to the different proportions of the components.

In order to further prove that the freeze-dried preparation prepared by the method has higher stability, the content of HPR before and after freezing is respectively detected for 3 samples prepared in each group of freeze-dried preparation samples prepared by application examples 1.7, 2.7, 3.7, 4.7, 5.7 and 6.7.

The detection method comprises the following steps:

the prepared freeze-dried preparation loaded with HPR is subjected to HPLC detection of HPR before freeze-drying and after freeze-drying respectively, and HPR chromatographic conditions are as follows: chromatographic column: c18 column (4.6 mm. Times.250 mm,5 μm); column temperature: 30 ℃, detection wavelength: 358nm, mobile phase: 98% acetonitrile+2% water, flow rate: 1.0mL/min; the procedure is as follows: and (5) performing isocratic elution.

Sample treatment: samples were taken before lyophilization and after lyophilization, respectively, and were sonicated with 98% acetonitrile+2% water as the mobile phase for 30min, filtered through 0.22um, and tested as set forth in Table 7 below.

TABLE 7 HPLC detection of hydroxy pinacolin retinoic acid ester content before and after lyophilization

As can be seen from table 7 and fig. 14, the lyophilized preparation prepared by the present application has high stability, and the retention rate of HPR before and after lyophilization of the lyophilized preparation prepared by application example 1.7 can reach 96.46%, and since the lyophilized preparations prepared by application examples 2.7, 3.7, 4.7, 5.7 and 6.7 are all lyophilized preparations within the maximum load range, the retention rates of HPR before and after lyophilization are equivalent to the values of application example 1.7, both of which are above 95%, which indicates that the stability of the active ingredient of the lyophilized preparation prepared by the present application is good before and after lyophilization.

In addition, the content of retinol palmitate before and after freezing was examined for the freeze-dried preparation samples prepared in application examples 1.6, 2.6, 3.6, 4.6, 5.6 and 6.6, respectively, for 3 samples prepared per group.

The detection method comprises the following steps:

the prepared freeze-dried system loaded with the retinol palmitate is subjected to HPLC detection of the retinol palmitate after freeze-drying and high-temperature treatment at 40 ℃ for 2 weeks after freeze-drying, and the chromatographic conditions of the retinol palmitate are as follows: chromatographic column: c18 column (4.6 mm. Times.250 mm,5 μm); column temperature: 30 ℃, detection wavelength: 325nm, mobile phase: 45% isopropanol +55% methanol, flow rate: 1.0mL/min; the procedure is as follows: and (5) performing isocratic elution.

Sample treatment: after freeze-drying and after 2 weeks of high temperature treatment at 40 ℃ after freeze-drying, samples were taken for HPLC detection, ultrasonic extraction was performed for 30min by dissolution with mobile phase 45% isopropanol+55% methanol, filtration was performed at 0.22um, and the detection results of the loading detection are shown in Table 8 below.

Table 8: HPLC detection result of retinol palmitate content after freeze-drying and 40 ℃ high temperature treatment for 2 weeks

As can be seen from the above Table 8 and FIG. 15, the lyophilized preparation prepared by the present application has high stability, and the retention rate of retinol palmitate after lyophilization and after 2 weeks of high temperature treatment at 40℃in the lyophilized preparation prepared by application example 1.6 can reach 95.41%, and since the lyophilized preparations prepared by application examples 2.6, 3.6, 4.6, 5.6 and 6.6 are all lyophilized preparations within the maximum load range, the retention rate of retinol palmitate after lyophilization and after 2 weeks of high temperature treatment at 40℃in the lyophilized preparation is equivalent to the value of retinol palmitate after application example 1.6, which is 95% or more, indicating that the lyophilized preparation prepared by the present application has good high temperature stability and is suitable for long-term storage.

Application example 7 freeze-dried preparation and preparation method thereof

The composite material comprises the following components in parts by weight:

the preparation method of the freeze-dried preparation comprises the following steps: (1) Uniformly mixing glycine, trehalose, hydrolyzed xylan, mannitol, rhamnose, xanthan gum and anthocyanin with the formula dosage, and sieving with a 150-mesh sieve to obtain a mixture A;

(3) Heating and melting the rest other components, adding into the mixture B in the step (2), and emulsifying by ultrasonic (at 40 ℃ C., 300W,30 min) to obtain a mixture C;

(4) Immersing the freeze-drying system carrying other components obtained in the step (3) into alginate fiber or silk membrane cloth until the membrane cloth is full of liquid, and then performing freeze-drying treatment to obtain the freeze-drying mask preparation.

Stage one: slowly heating the temperature in the freeze-drying bin to-30 ℃ within 1 hour, keeping the vacuum degree below 1 Pa, and keeping for 3 hours;

step two, slowly heating the temperature in the freeze-drying bin to-10 ℃ within 1 hour, and keeping the vacuum degree at 25-35 Pa for 4 hours;

and step three, slowly raising the temperature to 0 ℃ in a freeze-drying bin within 1 hour, and keeping the vacuum degree at 35-45 Pa for 1 hour.

stage one: the temperature in the freeze-drying bin reaches 5-15 ℃ within 1 hour, the vacuum degree is 45-55 Pa, and the temperature is kept for 2 hours;

stage two: the temperature in the freeze drying bin reaches 15-25 ℃ within 1 hour, the vacuum degree is reduced to 35-45 Pa, and the temperature is kept for 1 hour.

According to the application embodiment, the freeze-drying composition of the embodiment 5 can be used for realizing freeze-drying of various components, as shown in fig. 16, the obtained freeze-drying system loaded with other components is immersed in a silk membrane cloth substrate, so that freeze-drying can be realized, 20-25mL of water is added, and rehydration can be realized within a few seconds, and the use is convenient.

Lyophilized preparation prepared from comparative examples 1.1-1.8 and its preparation method

The composite material comprises the following components in parts by weight:

the preparation method of the freeze-dried preparation described in comparative example 1.1 is applied:

(1) Taking mannitol, xanthan gum and anthocyanin which are used in the formula, uniformly mixing, and sieving with a 150-mesh sieve to obtain a mixture A;

(2) Adding animal source butter into the mixture A in the step (1) after being melted by water-proof heating, and emulsifying by ultrasonic (at 40 ℃ and 300W for 30 min) to obtain a mixture B; shaping the mixture B to obtain a shaped mixture B;

(3) The mixture B obtained in the step (2) after shaping is placed in a low-temperature environment for freeze-drying treatment to obtain the cosmetic freeze-dried preparation

The preparation methods described in comparative examples 1.2 to 1.8 were the same as those of comparative example 1.1 except that the components added in step (2) were different.

Detection result:

and (3) effect verification: as can be seen from fig. 17B, when comparative examples 1.1, 1.2, 1.3, 1.6 and 1.7 were used, butter, shea butter, hawaii butter, retinol palmitate and HPR were loaded, respectively, and after ultrasonic emulsification, they were suspended as small particles of oil droplets on the surface of the liquid, were not emulsified, were dispersed as two phases, and were not freeze-dried subsequently.

As can be seen from FIG. 17A, the comparative examples 1.4, 1.5, and 1.8 were applied to completely dissolve and to form clear and transparent purple solutions, which were freeze-dried, and the freeze-dried pellets were collapsed and could not be molded during the freeze-drying process.

Comparative example 2 was used

With the formulation disclosed in example 1 of chinese patent CN111632005a, butter with a solid content of 10% was loaded in step (S23), but as can be seen from fig. 18, oil droplets float on the surface after loading with 10% butter, and were not emulsified, and lyophilization was not performed.

Detection result:

comparative example 3 was used

With the formulation disclosed in example 2 of chinese patent CN112315821a, then 10% hawaii fruit oil is loaded in the solid component in step (3), but as can be seen from fig. 19, after 10% hawaii fruit oil is loaded, oil drops float on the surface, and cannot be emulsified, and freeze-drying treatment cannot be performed.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof.

The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein. Any reference sign in a claim should not be construed as limiting the claim concerned.

Furthermore, it should be understood that although the present disclosure describes embodiments, not every embodiment is provided with a separate embodiment, and that this description is provided for clarity only, and that the disclosure is not limited to the embodiments described in detail below, and that the embodiments described in the examples may be combined as appropriate to form other embodiments that will be apparent to those skilled in the art.

Claims

1. A lyophilized composition comprising a scaffold molecule and a liposome entrapped therein, characterized in that: the freeze-dried composition comprises the following components in percentage by weight: 15% of glycine, 15% of trehalose, 15% of hydrolyzed xylan, 15% of mannitol, 15% of rhamnose, 6.25% of soybean lecithin and 18.75% of hydrogenated lecithin; wherein the skeleton molecule is a mixture of glycine, trehalose, hydrolyzed xylan, mannitol and rhamnose, and the liposome is embedded into a mixture of soybean lecithin and hydrogenated lecithin.

2. Use of a lyophilized composition according to claim 1 for the preparation of a lyophilized formulation, said lyophilized formulation being a cosmetic lyophilized formulation.

3. A cosmetic lyophilized formulation characterized by: comprises the freeze-dried composition containing skeleton molecules and liposome embedding as claimed in claim 1 and one or more of grease components, polyalcohol components, unstable and easily-decomposed components, easily-gelled components after drying, vitamins, thickeners and active ingredients;

wherein the grease component is selected from animal-derived grease and plant-derived grease; the animal-derived oil is one or more of butter, mink oil, egg yolk oil, lanolin oil and squalane; the vegetable oil is one or more selected from shea butter, hawaii fruit oil, jojoba seed oil, olive oil, coconut oil, castor oil, cotton seed oil, soybean oil, sesame oil, almond oil, peanut oil, corn oil, rice bran oil, tea seed oil, sea buckthorn oil, avocado oil, chestnut oil, walnut oil and cocoa butter; the polyalcohol component is selected from glycerol or/and propylene glycol; the unstable and easily-decomposed component is selected from retinol palmitate and/or hydroxy pinacolone retinol sulfonate; the components which are easy to form gel after being dried are selected from glass color factors; the vitamin is vitamin C and its salt; the thickening agent is selected from microbial glue; the active ingredients are one or more of adenosine, tocopheryl acetate, superoxide dismutase, sodium hyaluronate, anthocyanin, hydrolyzed ginsenoside, centella asiatica extract and ganoderma lucidum extract.

4. A method of preparing a cosmetic lyophilized formulation as defined in claim 3, wherein: the method comprises the following steps:

step three, slowly raising the temperature to about 0 ℃ in a freeze-drying bin within 1 hour, and keeping the vacuum degree at 35-45 Pa for 1-2 hours;