CN114917134A

CN114917134A - Freeze-dried composition containing skeleton molecules and liposome embedding and application thereof

Info

Publication number: CN114917134A
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: 2022-08-19
Anticipated expiration: 2042-06-07
Also published as: CN114917134B

Abstract

The invention discloses a freeze-dried composition containing a skeleton molecule and a liposome embedding and application thereof, relating 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. According to the invention, the skeleton molecules and the liposome are embedded and matched with each other, so that the freeze-dried preparation formed by the skeleton molecules and the liposome can be used for preparing components which are difficult to freeze dry conventionally, such as high-loading capacity, high-compatibility freeze-dried oils and fats, polyols, easily-decomposed unstable active substances, easily-formed colloids after drying and the like, and has the advantages of high loading capacity of effective components, 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 embedded by skeleton molecules and liposome and application thereof.

Background

The face skin care product is a cosmetic for face, which contains precious functional components such as plant extracts and the like, so that the skin care product has the effects of preventing aging, resisting wrinkles, moisturizing, whitening, removing freckles and the like. The conventional facial skin care products generally contain unstable active substances, thereby affecting the long-term preservation of the skin care products, and need emulsification due to the lipophilicity of the skin to better preserve and deliver the active substances, so a large amount of emulsifying agents, skin feel conditioning agents and the like are needed to be added for improving the stability of the products, and the conventional facial skin care products are mostly in the form of bottled liquid, in order to prevent the breeding of bacteria, preservatives are needed to be added, and the problems of inconvenient carrying exist, and the repeated switching has the risk of cross contamination, which can cause damage to the skin, so the ultra-low temperature vacuum freeze drying technology of the cosmetics comes along.

Nowadays, many frozen cosmetics such as freeze-dried facial mask, freeze-dried cosmetic powder and the like are available on the market, and they are prepared by freeze-drying technology. The freeze drying technology is to freeze the matter containing great amount of water into solid through lowering the temperature and vacuum condition to sublimate the solid water directly while the matter remains in the frozen ice rack for storing the active components of biological matter fully.

For example, patent CN111632005A discloses a polypeptide lyophilized powder mask composition and its preparation and application methods. The polypeptide freeze-dried powder mask composition consists of polypeptide freeze-dried powder, freeze-dried solvent, mask liquid and mask cloth. The freeze-drying technology is adopted to store the oligopeptide-5, the oligopeptide-1 and other materials 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 framework is provided for the material in the freeze-drying process, and the material is shaped. Said invention not only retains the biological active substance and heat-sensitive substance 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 has the advantages of high active component, and can greatly retain the biological active substance and heat-sensitive substance in the material, however, the addition of preservative in the above-mentioned polypeptide freeze-dried powder facial mask has a certain irritation to skin, and its safety is reduced.

Another patent CN112315821A discloses a hydrolyzed protein liposome and its preparation method. The hydrolyzed protein liposome consists of a cosmetic active component, a liposome framework component, an object component and a freeze-drying protective agent. The liposome is used as a carrier of the hydrolyzed keratin, and the lactose, mannitol and trehalose are used as freeze-drying protective agents, so that the transdermal absorption of the hydrolyzed keratin is promoted, and the concentration of active substances is continuously improved in the process of contacting with the skin. However, since the freeze-dried mask system contains only an aqueous substance and does not freeze-dry an oil-and-fat component, the obtained mask has insufficient skin feel and poor experience.

Therefore, there is an urgent need for a freeze-dried preparation which is free of preservatives and chemical auxiliaries, has a light weight without moisture, and can freeze-dry an oil-and-fat component.

Disclosure of Invention

Aiming at the defects of the prior technical scheme, the invention aims to provide a freeze-dried preparation which has high loading of effective components, high compatibility of oily components and aqueous components, multiple dimensionality of skin feel, good experience feel, low moisture content, light weight and no need of adding chemical synthetic oil, skin feel regulator, preservative and the like.

In order to achieve the purpose, the technical scheme is as follows:

the first purpose of the invention is to provide a freeze-dried composition containing a skeleton molecule and a liposome embedding, wherein the freeze-dried composition 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 skeleton molecules and 25-45% of liposome embedding.

Still preferably, the lyophilized composition comprises the following components in percentage by weight: 75% of skeleton molecules and 25% of liposome embedding.

Wherein said backbone molecule comprises an amino acid;

the amino acid is selected from 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 amphipathy of acid and alkali, so that the pH of the solution can be kept stable in the processes of low-temperature storage and freeze drying of biological products, and the aim of protecting active components is fulfilled; the crystalline glycine can raise the collapse temperature of the finished product and prevent the protein medicine from being damaged due to collapse.

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

the trehalose is a glucose dimer, is stable non-reducing disaccharide, is widely used for freeze-drying preservation, is stable in biological performance of a freeze-dried sample as a protective agent, is long in preservation time, is the best protective agent in the low-temperature biological field, is small in molecular weight, is easy to fill in gaps inside macromolecules, effectively limits structural change inside the macromolecules, avoids active matter inactivation, is high in glass transition temperature, and can effectively prevent disintegration and collapse. Mannitol is a polyhydroxy compound, can form a loose and firm uniform framework as a carrier, and also can be used as a permeability regulator and a protein freeze-drying protective agent. Hydrolyzed xylan, rhamnose and other sugars are the most common and widely used freeze-drying protective agents, and play a role in protecting active substances in each freeze-drying stage.

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

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

The liposome embedding is selected from 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 (an oil phase and a water phase) in the other phase, and the invention adopts soybean lecithin and hydrogenated lecithin with the mass ratio of 1:3 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 of the composition, the embedded mass ratio of the skeleton molecules and the liposome, particularly the mass ratio of glycine, trehalose, hydrolyzed xylan, mannitol and rhamnose and the mass ratio of soybean lecithin and hydrogenated lecithin are reasonably controlled, so that the obtained composition can better freeze-dry one or more of oil and fat components, polyol components, unstable and easily decomposed components, dried easily-gelatinized components, vitamins, thickeners and active ingredients.

Therefore, another object of the present application is to provide a freeze-dried cosmetic preparation, which comprises the above freeze-dried composition, and further comprises one or more of an oil component, a polyol component, an unstable and easily degradable component, a component which is easily gelled after drying, a vitamin, a thickener and an active ingredient;

the oil and fat component is selected from animal source oil and fat and vegetable source oil and fat; the animal source oil is selected from one or more of butter, mink oil, egg yolk oil, lanolin oil and squalane; the vegetable source oil is selected from one or more of shea butter, macadamia nut oil, jojoba seed oil, olive oil, coconut oil, castor oil, cottonseed oil, soybean oil, sesame oil, almond oil, peanut oil, corn oil, rice bran oil, tea seed oil, sea buckthorn oil, avocado oil, Chinese chestnut oil, walnut oil and cocoa butter; the polyalcohol component is selected from glycerol or/and propylene glycol; the unstable easily decomposable component is selected from retinol palmitate and/or hydroxy pinacolone retinoic acid ester; the easy-to-gel component after drying is selected from a vitreous chromogen.

The vitamin is vitamin C and salt thereof; the thickening agent is selected from one or more of microbial gum, xanthan gum and sclerotium rolfsii gum; the active ingredient is one or more of adenosine, tocopherol acetate, superoxide dismutase, sodium hyaluronate, anthocyanin, hydrolyzed ginsenosides, centella asiatica extract and Ganoderma lucidum extract.

The third purpose of the invention is to provide a preparation method of the cosmetic freeze-dried preparation, which comprises the following steps:

(1) weighing framework molecules, vitamins, a thickening agent and active ingredients according to the formula, uniformly mixing and sieving to obtain a mixture A;

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

(3) loading one or more of oil components, polyalcohol components and unstable and easily degradable components and easy-to-gel components 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) performing 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, sublimating and analyzing and drying;

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

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

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

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

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

The analysis drying is divided into two stages, and the operation is specifically as follows:

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

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

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

firstly, components which are difficult to freeze dry conventionally, such as high-loading and high-compatibility freeze-dried oil and fat, polyhydric alcohols, unstable active substances which are easy to decompose, colloidal saccharides which are easy to gel after drying, and the like can be obtained;

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 light, and the carrying and the long-distance transportation are convenient.

Drawings

FIG. 1 is a drawing showing the state of a cut surface and a reconstituted state of a lyophilized preparation prepared by 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 standing at 40 ℃ for 2 weeks and at room temperature;

FIG. 3 is a drawing showing the state of a cut surface and a reconstituted state of a lyophilized preparation prepared by using 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 standing at 40 ℃ for 2 weeks;

FIG. 5 is a drawing showing the state of a cut surface and a reconstituted state of a lyophilized preparation prepared by 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 standing at 40 ℃ for 2 weeks and at room temperature;

FIG. 7 is a graph showing the knife-cut state and reconstitution state 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 standing at 40 ℃ for 2 weeks and at room temperature;

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

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 a high temperature of 40 ℃ and at room temperature for 2 weeks;

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

FIG. 12 is a graph showing the effect of the lyophilized preparation prepared in examples 6.1 to 6.8 after standing at 40 ℃ for 2 weeks;

FIG. 13 is a graph comparing the maximum loading of each component in the lyophilized formulations prepared using examples 1-6;

FIG. 14 is a hydroxy pinacolone retinoic acid ester (HPR) HPLC chromatogram of a lyophilized formulation prepared in application example 1.7 before and after lyophilization;

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

FIG. 15 is a HPLC plot of retinol palmitate content after lyophilization and after 2 weeks of high temperature treatment at 40 ℃ after lyophilization of the lyophilized formulation prepared in application example 1.6;

wherein A is an HPLC (high performance liquid chromatography) spectrum of a retinol palmitate standard substance, B is an HPLC spectrum of a freeze-dried preparation, and C is an HPLC spectrum of the freeze-dried preparation after being treated at the high temperature of 40 ℃ for 2 weeks;

fig. 16 is a view showing a state after application of the lyophilized mask preparation prepared in example 7 and reconstitution;

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

FIG. 18 is a diagram showing the dissolution state of a formulation described in comparative example 2;

FIG. 19 is a graph showing the dissolution state of the formulation of comparative example 3.

Detailed Description

The features mentioned above in the description, or the features mentioned in the embodiments, may be combined arbitrarily. All the features disclosed in this specification may be combined in any suitable manner and each feature disclosed in this specification may be replaced by an alternative feature serving the same, equivalent or similar purpose. Thus, unless expressly stated otherwise, the features disclosed are merely generic examples of equivalent or similar features.

The invention will be further illustrated with reference to the following specific examples. These examples are intended to illustrate the invention and are not intended to limit the scope of the invention. The following examples are conducted under conditions specified, usually according to conventional conditions or according to conditions recommended by the manufacturer. All percentages and fractions are by weight unless otherwise specified.

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 invention. The preferred embodiments and materials described herein are exemplary only.

The purchase producer and the model of raw materials and instrument that use in this application experimentation are as follows:

examples 1-6A lyophilized composition comprising a matrix molecule and an entrapped liposome

Comprises the following components in percentage by weight:

application example 1.1-1.8A lyophilized preparation and a method for preparing the same

The paint comprises the following components in parts by weight:

the method comprises the following steps:

(1) weighing glycine, xanthan gum and anthocyanin according to the formula, uniformly mixing, and sieving by a 150-mesh sieve to obtain a mixture A;

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

(3) heating and melting animal source butter, adding the animal source butter into the mixture B in the step (2), and emulsifying by ultrasonic waves (40 ℃, 300W and 30min) 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, sublimating and analyzing and drying;

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

The same procedures as those conducted in practical example 1.1 were conducted except that the components added in step (3) were different according to the production methods described in practical examples 1.2 to 1.8.

And (3) detection results:

as shown in the attached figure 1, the freeze-dried samples are prepared according to the application examples 1.1-1.8, and the obtained freeze-dried spheres are well formed, complete in shape and smooth in surface; the knife cutting surface layer is in a laminated shape; the emulsion is fast to rehydrate and can be dissolved within 2 minutes, and the emulsion has a good emulsifying effect.

Test example 1-1 detection of emulsion value OD500

According to the evaluation and detection method of the emulsification stability of the protein emulsion, namely the emulsification activity index, the light absorption value of a sample at 500nm is detected, and the emulsification and the stability of the sample can be evaluated in a correlation manner; the OD500 values of the lyophilized formulations prepared in examples 1.1 to 1.8 were measured by redissolving in water after lyophilization, and the results are shown in table 1 below.

TABLE 1

	OD500 value of preparation	Rehydration OD500 value after freeze drying	Variation 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％

And (4) conclusion: the OD500 values of the freeze-dried preparation prepared in the application examples 1.1 to 1.8 and the redissolved water after freeze-drying are within +/-5%, which shows that the emulsification effect and the emulsification stability are better after the redispersion.

Test examples 1-2 high temperature stability test

Referring to FIG. 2, using the lyophilized preparations prepared in examples 1.1 to 1.8, 3 vials each were put in a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and were taken out after 2 weeks, and compared with a sample after being left at room temperature for 2 weeks, the color and morphology were not changed, indicating high temperature stability.

Test examples 1 to 3 detection of the highest load amount

The determination criteria for the maximum load amount are: the freeze-dried balls are well formed, and the surfaces of the freeze-dried balls are smooth and have no liquid; the emulsion is an emulsion after rehydration, has no oil drops and is not layered.

The highest loading of the lyophilized composition described in example 1 was examined, and the maximum loading of the lyophilized composition described in example 1 for animal-derived butter was 10%; the maximum loading for shea butter was 1%; the maximum loading for macadamia nut oil was 10%; the maximum loading for glycerol was 1%; the maximum loading for propylene glycol was 1%; the maximum loading for retinol palmitate was 1%; the maximum loading for HPR is 1%; the maximum loading for the boscalid was 5%.

Application example 2.1-2.8A lyophilized preparation and a method for preparing the same

The paint comprises the following components in parts by weight:

the method comprises the following steps:

(1) weighing glycine, trehalose, xanthan gum and anthocyanin according to formula dosage, uniformly mixing, and sieving by a 150-mesh sieve to obtain a mixture A;

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

The same procedures as those conducted in practical example 2.1 were conducted except that the components added in step (3) were different according to the production methods described in practical examples 2.2 to 2.8.

And (3) detection results:

as shown in fig. 3, the freeze-dried samples prepared according to the application examples 2.1-2.8 have good molding, complete shape and smooth surface; the knife cutting surface layer is in a laminated shape; the emulsion is fast to rehydrate and can be dissolved within 2 minutes, and the emulsion has a good emulsifying effect.

Test example 2-1 detection of emulsion value OD500

According to the evaluation and detection method of the emulsification stability of the protein emulsion, namely the emulsification activity index, the light absorption value of a sample at 500nm is detected, and the emulsification and the stability of the sample can be evaluated in a correlation manner; the OD500 values of the lyophilized formulations prepared in examples 2.1 to 2.8 were measured by redissolving in water after lyophilization, and the results are shown in table 2 below.

TABLE 2

	OD500 value of preparation	Rehydration OD500 value after freeze drying	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％

And (4) conclusion: the OD500 values of the freeze-dried preparation prepared in the application examples 2.1-2.8 and the redissolved water after freeze drying are within +/-5%, which shows that the emulsification effect and the emulsification stability are better after the redissolution.

Test example 2-2 high temperature stability test

Referring to FIG. 4, using the lyophilized preparations prepared in examples 1.1 to 1.8, 3 vials each were put in a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and were taken out after 2 weeks, and compared with a sample after being left at room temperature for 2 weeks, the color and morphology were not changed, indicating high temperature stability.

Test examples 2-3 maximum load detection

The determination criteria for the maximum load amount are: the freeze-dried balls are well formed, and the surfaces of the freeze-dried balls are smooth and have no liquid; the emulsion is emulsion after rehydration, has no oil drop and is not layered.

The highest loading of the lyophilized composition described in example 2 was examined, and the maximum loading of the lyophilized composition described in example 2 for animal-derived butter was 15%; the maximum loading for shea butter was 5%; the maximum loading for macadamia nut oil was 15%; the maximum loading for glycerol was 3%; the maximum loading for propylene glycol was 3%; the maximum loading for retinol palmitate was 3%; the maximum loading for HPR is 3%; the maximum loading for the boscalid was 10%.

Application example 3.1-3.8A lyophilized preparation and a method for preparing the same

The paint comprises the following components in parts by weight:

the preparation method of the lyophilized preparation described in application example 3.1: the method comprises the following steps:

(1) weighing glycine, trehalose, mannitol, rhamnose, xanthan gum and anthocyanin according to formula dosage, uniformly mixing, and sieving by a 150-mesh sieve to obtain a mixture A;

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

the pre-freezing operation comprises the following steps: starting a freeze-drying device, enabling the temperature of a cold well to reach below-45 ℃ in about 40 minutes from room temperature, and keeping for 2 hours;

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

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

And (3) detection results:

as shown in fig. 5, the freeze-dried samples prepared according to the application examples 3.1-3.8 have good molding, complete shape and smooth surface; the knife cutting surface layer is in a laminated shape; the emulsion has fast rehydration and can be dissolved in 2 minutes to form emulsion with good emulsification effect.

Test example 3-1 detection of emulsion value OD500

According to the evaluation and detection method of the emulsification stability of the protein emulsion, namely the emulsification activity index, the light absorption value of a sample at 500nm is detected, and the emulsification and the stability of the sample can be evaluated in a correlation manner; the OD500 values of the lyophilized formulations prepared in examples 3.1 to 3.8 were measured by reconstitution in water after lyophilization, and the results are shown in table 3 below.

TABLE 3

	OD500 value of preparation	Rehydration OD500 value after freeze drying	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％

And (4) conclusion: the OD500 values of the freeze-dried preparation prepared in application examples 3.1-3.8 and the redissolved in water after freeze-drying are within +/-5%, which shows that the emulsification effect is better and the emulsification stability is better after the redissolution.

Test example 3-2 high temperature stability test

Referring to FIG. 6, using the lyophilized preparations prepared in examples 1.1 to 1.8, 3 vials each were put in a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and were taken out after 2 weeks, and compared with a sample after being left at room temperature for 2 weeks, the color and morphology were not changed, indicating high temperature stability.

Test examples 3-3 maximum load detection

The highest loading of the lyophilized composition described in example 3 was examined, and the maximum loading of the lyophilized composition described in example 3 for animal-derived butter was 15%; the maximum loading for shea butter was 5%; the maximum loading for macadamia nut oil was 15%; the maximum loading for glycerol was 3%; the maximum loading for propylene glycol was 3%; the maximum loading for retinol palmitate was 3%; the maximum loading for HPR is 3%; the maximum loading for the boscalid was 10%.

Application example 4.1-4.8A lyophilized preparation and a method for preparing the same

The paint comprises the following components in parts by weight:

the preparation method of the lyophilized preparation described in application example 4.1: the method comprises the following steps:

(1) weighing glycine, trehalose, hydrolyzed xylan, mannitol, xanthan gum and anthocyanin according to the formula, uniformly mixing, and sieving by a 150-mesh sieve to obtain a mixture A;

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

The same procedures as those conducted in practical example 4.1 were conducted except that the components added in step (3) were different according to the production methods described in practical examples 4.2 to 4.8.

And (3) detection results:

as shown in fig. 7, the freeze-dried samples prepared according to the application examples 4.1-4.8 have good molding, complete shape and smooth surface; the knife cutting surface layer is in a laminated shape; the emulsion has fast rehydration and can be dissolved in 2 minutes to form emulsion with good emulsification effect.

Test example 4-1 detection of emulsion value OD500

According to the evaluation and detection method of the emulsification stability of the protein emulsion, namely the emulsification activity index, the light absorption value of a sample at 500nm is detected, and the emulsification and the stability of the sample can be evaluated in a correlation manner; the lyophilized formulations prepared in examples 4.1 to 4.8 were applied and reconstituted in water after lyophilization to measure the OD500 values thereof, and the results are shown in Table 4 below.

TABLE 4

	OD500 value of preparation	Rehydration OD500 value after freeze drying	Variation 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％

And (4) conclusion: the OD500 values of the freeze-dried preparation prepared in application examples 4.1-4.8 and the redissolved in water after freeze-drying are within +/-5%, which shows that the emulsification effect is better and the emulsification stability is better after the redissolution.

Test example 4-2 high temperature stability test

Referring to FIG. 8, using the lyophilized preparations prepared in examples 1.1 to 1.8, 3 vials each were put in a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and were taken out after 2 weeks, and compared with a sample after being left at room temperature for 2 weeks, the color and morphology were not changed, indicating high temperature stability.

Test examples 4-3 maximum load detection

The highest loading of the lyophilized composition described in example 4 was examined, and the maximum loading of the lyophilized composition described in example 4 for animal-derived butter was 30%; the maximum loading for shea butter was 5%; the maximum loading for macadamia nut oil was 30%; the maximum loading for glycerol was 5%; the maximum loading for propylene glycol was 5%; the maximum loading for retinol palmitate was 5%; the maximum loading for HPR is 5%; the maximum loading for the boscalid was 20%.

Application example 5.1-5.8A lyophilized preparation and a method for preparing the same

The paint comprises the following components in parts by weight:

the preparation method of the lyophilized preparation described in application example 5.1: the method comprises the following steps:

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

The same procedures as those conducted in practical example 5.1 were conducted except that the components added in step (3) were different according to the methods of practical examples 5.2 to 5.8.

And (3) detection results:

as shown in fig. 9, the freeze-dried samples prepared according to the application examples 5.1-5.8 have good molding, complete shape and smooth surface; the knife cutting surface layer is in a laminated shape; the emulsion has fast rehydration and can be dissolved in 2 minutes to form emulsion with good emulsification effect.

Test example 5-1 emulsion value OD500 measurement

According to the evaluation and detection method of the emulsification stability of the protein emulsion, namely the emulsification activity index, the light absorption value of a sample at 500nm is detected, and the emulsification and stability can be evaluated in a correlation manner; the OD500 values of the lyophilized formulations prepared in examples 5.1 to 5.8 were measured by redissolving in water after lyophilization, and the results are shown in table 5 below.

TABLE 5

And (4) conclusion: the OD500 values of the freeze-dried preparation prepared in the application examples 5.1 to 5.8 and the redissolved water after freeze-drying are within +/-5%, which shows that the emulsification effect and the emulsification stability are better after the redispersion.

Test example 5-2 high temperature stability test

Referring to FIG. 10, using the lyophilized preparations prepared in examples 1.1 to 1.8, 3 vials each were put in a high temperature treatment at 40 ℃ for 2 weeks after lyophilization, and were taken out after 2 weeks, and compared with a sample after being left at room temperature for 2 weeks, the color and morphology were not changed, indicating high temperature stability.

Test examples 5-3 maximum load detection

The highest loading of the lyophilized composition described in example 5 was examined, and the maximum loading of the lyophilized composition described in example 5 for animal-derived butter was 50%; the maximum loading for shea butter was 10%; the maximum loading for macadamia nut oil was 50%; the maximum loading for glycerol was 10%; the maximum loading for propylene glycol was 10%; the maximum loading for retinol palmitate was 10%; the maximum loading for HPR is 10%; the maximum loading for the boscalid was 30%.

Application example 6.1-6.8A lyophilized preparation and a method for preparing the same

The paint comprises the following components in parts by weight:

the preparation method of the lyophilized preparation described in application example 6.1: the method comprises the following steps:

(3) heating and melting animal source butter, adding the 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;

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

The same procedures as those conducted in practical example 6.1 were conducted except that the components added in step (3) were different according to the production methods described in practical examples 6.2 to 6.8.

And (3) detection results:

as shown in fig. 11, the freeze-dried samples prepared according to the application examples 6.1-6.8 have good molding, complete shape and smooth surface; the knife cutting surface layer is in a laminated shape; the emulsion is fast to rehydrate and can be dissolved within 2 minutes, and the emulsion has a good emulsifying effect.

Test example 6-1 emulsion value OD500 measurement

According to the evaluation and detection method of the emulsification stability of the protein emulsion, namely the emulsification activity index, the light absorption value of a sample at 500nm is detected, and the emulsification and stability can be evaluated in a correlation manner; the OD500 values of the lyophilized formulations prepared in examples 6.1 to 6.8 were measured using reconstitution in water after lyophilization, and the results are shown in table 6 below.

TABLE 6

	OD500 value of preparation	Rehydration OD500 value after freeze drying	Variation 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％

And (4) conclusion: the OD500 values of the freeze-dried preparation prepared in application examples 6.1-6.8 and the redissolved in water after freeze-drying are within +/-5%, which shows that the emulsification effect is better and the emulsification stability is better after the redissolution.

Test example 6-2 high temperature stability test

Referring to FIG. 12, using the lyophilized preparations prepared in examples 1.1 to 1.8, 3 bottles of each of the lyophilized preparations were put at 40 ℃ for 2 weeks, and 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 6 to 3 maximum load amount detection

The highest loading of the lyophilized composition described in example 6 was examined, and the maximum loading of the lyophilized composition described in example 6 for animal-derived butter was 30%; the maximum loading for shea butter was 5%; the maximum loading for macadamia nut oil was 30%; the maximum loading for glycerol was 10%; the maximum loading for propylene glycol was 10%; the maximum loading for retinol palmitate was 5%; the maximum loading for HPR is 5%; the maximum loading for the boscalid was 30%.

As can be seen from fig. 13, the specific components embedded by the skeleton molecules and the liposomes have a great influence on the loading capacity of the oil components, the polyol components, the unstable and easily degradable components and the easily gelling components after drying, in example 1, glycine is used as the skeleton molecules, and soybean lecithin is embedded by the liposomes, so that the obtained freeze-dried composition has a small loading capacity on the oil components, the polyol components, the unstable and easily degradable components and the easily gelling components after drying; example 2 glycine and trehalose are used as framework molecules, and soybean lecithin and hydrogenated lecithin are used as liposomes for embedding, so that the load capacity of the obtained freeze-dried composition on oil components, polyol components, unstable and easily-degradable components and easily-gelling components after drying is improved in comparison with example 1, but is still lower; examples 3 and 4, various components are added, and the oil component, the polyol component and the unstable and easily degradable component of the obtained freeze-dried composition are obviously improved, and the loading capacity of the easily gelling component after drying is obviously improved in examples 1-2; example 5 by controlling the backbone molecules to 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 oil components, polyol components, unstable and easily-degradable components and easily-gelled components after drying can be obviously improved; the components of the lyophilized composition disclosed in example 6 were the same as in example 5, and the entrapping contents of the backbone molecules and the liposomes were the same as in example 5, but the loading amounts of the oil-and-fat component, the polyol component, the unstable and easily decomposable component, and the easily gellable component after drying were significantly reduced due to the different ratios among the components.

To further demonstrate the higher stability of the lyophilized formulations prepared in the present application, the HPR content was measured before and after freezing for the lyophilized formulation samples prepared in application examples 1.7, 2.7, 3.7, 4.7, 5.7 and 6.7, respectively, for each of 3 samples prepared.

The detection method comprises the following steps:

the prepared lyophilized preparation loaded with HPR is subjected to HPLC detection of HPR before and after lyophilization respectively, and the chromatographic conditions of HPR are as follows: a 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: 1.0 mL/min; the procedure is as follows: and (3) carrying out isocratic elution.

Sample treatment: before freeze-drying and after freeze-drying, respectively sampling, dissolving with mobile phase 98% acetonitrile and 2% water, ultrasonic extracting for 30min, filtering for 0.22um, loading and detecting, and the detection result is shown in the following table 7.

TABLE 7 HPLC test results of hydroxy pinatone retinoic acid ester content before and after lyophilization

As can be seen from table 7 above and fig. 14, the lyophilized preparation prepared in the present application has high stability, and the retention rates of HPR before and after lyophilization of the lyophilized preparation prepared in application example 1.7 can reach 96.46%, and since the lyophilized preparations prepared in 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 value of application example 1.7, and are both above 95%, which indicates that the lyophilized preparation prepared in the present application has good stability of active ingredients before and after lyophilization.

In addition, with respect to the samples of the lyophilized preparations prepared in application examples 1.6, 2.6, 3.6, 4.6, 5.6 and 6.6, the retinol palmitate contents before and after freezing were measured for 3 samples prepared in each group.

The detection method comprises the following steps:

carrying out HPLC detection on the prepared freeze-dried system carrying the retinol palmitate after freeze-drying and high-temperature treatment at 40 ℃ for 2 weeks respectively, wherein the chromatographic conditions of the retinol palmitate are as follows: and (3) 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: 1.0 mL/min; the procedure is as follows: and (3) carrying out isocratic elution.

Sample treatment: after freeze drying and high temperature treatment at 40 ℃ for 2 weeks after freeze drying, sampling for HPLC detection, dissolving with mobile phase of 45% isopropanol and 55% methanol, ultrasonic extracting for 30min, filtering at 0.22um, and loading detection results are shown in Table 8 below.

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

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

Application example 7A lyophilized preparation and preparation method thereof

The paint 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 according to 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 treatment (40 ℃, 300W and 30min) to obtain a mixture C;

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

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

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

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

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

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

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

Application comparative example 1.1-1.8 freeze-dried preparation and preparation method thereof

The paint comprises the following components in parts by weight:

the preparation method of the lyophilized formulation described in comparative example 1.1 was applied:

(1) uniformly mixing mannitol, xanthan gum and anthocyanin according to the formula dosage, and sieving the mixture through a 150-mesh sieve to obtain a mixture A;

(2) heating and melting animal source butter in a water-proof manner, adding the melted animal source butter into the mixture A obtained in the step (1), and emulsifying by ultrasonic waves (40 ℃, 300W and 30min) to obtain a mixture B; then shaping the mixture B to obtain a shaped mixture B;

(3) placing the molded mixture B obtained in the step (2) in a low-temperature environment for freeze-drying treatment to obtain the freeze-dried cosmetic preparation

The same procedure as in comparative example 1.1 was carried out except that the components added in step (2) were different in the production processes described in comparative examples 1.2 to 1.8.

And (3) detection results:

effect verification: as can be seen from fig. 17B, the samples of comparative examples 1.1, 1.2, 1.3, 1.6, and 1.7, which were loaded with butter, shea butter, macadamia nut oil, retinol palmitate, and HPR, respectively, were not loaded, and after ultrasonic emulsification, the oil particles floated on the liquid surface as oil droplets, were not emulsified, and were in two dispersed phases, and thus, were not freeze-dried subsequently.

As can be seen from FIG. 17A, the freeze-dried pellets were freeze-dried in a clear and bright purple solution with complete dissolution using comparative examples 1.4, 1.5 and 1.8, and were collapsed and could not be molded during the freeze-drying process.

Comparative application example 2

With the formulation disclosed in example 1 of chinese patent CN111632005A, then, in step (S23), butter with a solid content of 10% was loaded, but as can be seen from fig. 18, after the butter with a solid content of 10% was loaded, oil droplets floated on the surface and were not emulsified, and thus, the freeze-drying treatment was not performed.

And (3) detection results:

comparative application example 3

By adopting the formula disclosed in example 2 of chinese patent CN112315821A, then the macadamia nut oil with a solid content of 10% is loaded in step (3), but as can be seen from fig. 19, after the macadamia nut oil with a solid content of 10% is loaded, oil droplets float on the surface, and are not emulsified, and thus the 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 attributes 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 description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.

Claims

1. A lyophilized composition comprising a scaffold molecule and a liposome entrapped therein, wherein: the binary freeze-dried composition comprises the following components in percentage by weight: 55-80% of skeleton molecules and 20-45% of liposome embedding.

2. The lyophilized composition of claim 1, wherein: comprises the following components in percentage by weight: 60-75% of skeleton molecules and 25-45% of liposome embedding.

3. The lyophilized composition according to claim 2, wherein: comprises the following components in percentage by weight: 75% of skeleton molecules and 25% of liposome embedding.

4. The lyophilized composition of claim 3, wherein: one or more of amino acid, trehalose, hydrolyzed xylan, mannitol and rhamnose;

the liposome is embedded into lecithin.

5. The lyophilized composition according to claim 4, wherein: the skeleton molecule is a mixture of glycine, trehalose, hydrolyzed xylan, mannitol and rhamnose;

the lecithin is soybean lecithin and/or hydrogenated lecithin.

6. The lyophilized composition according to claim 5, wherein: the skeleton molecules are glycine, trehalose, hydrolyzed xylan, mannitol and rhamnose in a mass ratio of 1:1:1: 1;

the lecithin is soybean lecithin and hydrogenated lecithin in a mass ratio of 1: 3.

7. Use of a lyophilized composition according to any one of claims 1-6 for the preparation of a lyophilized formulation, which is a cosmetic lyophilized formulation.

8. A cosmetic lyophilized preparation characterized by: comprising the binary freeze-dried composition as defined in any one of claims 1 to 6 and one or more of a fat component, a polyol component, an unstable and easily decomposable component, a component which is easily gelled after drying, a vitamin, a thickener and an active ingredient.

9. The lyophilized formulation for cosmetics according to claim 8, characterized in that: the oil component is selected from animal source oil and vegetable source oil; the animal source oil is selected from one or more of butter, mink oil, egg yolk oil, lanolin oil and squalane; the vegetable source oil is selected from one or more of shea butter, macadamia nut oil, jojoba seed oil, olive oil, coconut oil, castor oil, cottonseed oil, soybean oil, sesame oil, almond oil, peanut oil, corn oil, rice bran oil, tea seed oil, sea buckthorn oil, avocado oil, Chinese chestnut oil, walnut oil and cocoa butter; the polyalcohol component is selected from glycerol or/and propylene glycol; the unstable easily-decomposable component is selected from retinol palmitate and/or hydroxy pinacolone retinoic acid ester; the component which is easy to glue after drying is selected from vitronectin.

10. The method for producing a lyophilized cosmetic preparation according to any one of claims 8 to 9, characterized in that: the method comprises the following steps:

(1) weighing skeleton molecules, vitamins, a thickening agent and active ingredients in the formula amount, uniformly mixing and sieving to obtain a mixture A;

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

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