CN115634167B

CN115634167B - Liposome containing ferulic acid and derivative thereof, and preparation method and application thereof

Info

Publication number: CN115634167B
Application number: CN202211251631.1A
Authority: CN
Inventors: 畅通; 甄文超; 传雅萍; 王佳
Original assignee: Shaanxi Changxiang Pharmaceutical Co ltd
Current assignee: Shaanxi Changxiang Pharmaceutical Co ltd
Priority date: 2022-10-12
Filing date: 2022-10-12
Publication date: 2024-05-14
Anticipated expiration: 2042-10-12
Also published as: CN115634167A

Abstract

The invention discloses a liposome containing ferulic acid and derivatives thereof, and a preparation method and application thereof, and belongs to the technical field of cosmetics. The liposome containing ferulic acid and derivatives thereof is of a core-shell structure, the core of the liposome comprises ferulic acid and derivatives thereof, and the shell of the liposome comprises phytosterol, phospholipid, glyceride compounds and fatty acyl compounds. The liposome containing the ferulic acid and the derivatives thereof has uniform particle size, high dispersibility, excellent stability and embedding rate of more than 85 percent through the selection of raw materials and the improvement of the process. The ferulic acid and the derivatives thereof wrapped by the liposome have excellent stability and long-acting slow release effect, and the liposome is used for conveying the ferulic acid and the derivatives thereof from the epidermis of the skin to the inside of the skin, so that the beneficial effects of whitening, antioxidation, anti-inflammation, acne removal and the like of the ferulic acid and the derivatives thereof are better exerted, and the liposome can be applied to the field of cosmetics.

Description

Liposome containing ferulic acid and derivative thereof, and preparation method and application thereof

Technical Field

The invention relates to the technical field of cosmetics, in particular to a liposome containing ferulic acid and derivatives thereof, and a preparation method and application thereof.

Background

The phenolic hydroxyl groups of the ferulic acid and the derivatives thereof have obvious scavenging effect on peroxy free radicals and hydroxy free radicals, reduce the risk of skin function injury caused by oxidation reaction of biological tissue membranes, effectively inhibit the activity of tyrosinase, inhibit the generation of melanin and achieve the effects of whitening, resisting oxidation and delaying skin aging; the styryl structure has the effect of antagonizing endothelin, plays roles in protecting vascular endothelium, enhancing curative effect, expanding treatment range, reducing toxic and side effects, achieving anti-inflammatory effect, and relieving red and inflamed vaccinia muscle, and the derivative has the advantages and better stability, but is still easily influenced by factors such as light, heat, oxidization and the like to easily lose bioactivity.

Liposomes (liposome) are artificial membranes, which are closed vesicles of spheroid bilayer lipid molecules with diameters of 20nm to tens of microns, in which phospholipid molecules are dispersed in water to form an encapsulated aqueous phase. The liposome has many unique physical and chemical properties, is formed by phospholipid spontaneously in water, has the characteristics of no toxicity, no immunogenicity, degradability, slow release, targeting and the like in human body, can be widely used as a drug carrier for transgenic technology or for preparing drugs, and can be delivered into the inside of cells or interstitial spaces by utilizing the characteristic that the liposome can be fused with cell membranes. Liposomes can also target the skin and deliver the active substance into the skin, exerting better effects, often in the cosmetic field.

Conventional liposome preparation methods include dissolving the multi-layer or bilayer constituent components with a volatile organic solvent mixture or mixtures, then evaporating the organic solvent without phase separation, and retaining the encapsulate and liposome components, wherein during the preparation process, a solubilizing agent is usually added, which can cause foaming during the product use process, resulting in poor use experience, and during transportation and storage, the liposome is easily solubilized and ruptured, and the active ingredient is easily separated.

In addition, less research is carried out on the liposome of the ferulic acid and the derivative thereof, so that the liposome which is used for wrapping the ferulic acid and the derivative thereof and has good stability, high encapsulation efficiency and uniform and controllable particle size is researched, and the beneficial effects of the ferulic acid and the derivative thereof can be exerted to the maximum extent.

Disclosure of Invention

The invention mainly aims to provide a liposome containing ferulic acid and derivatives thereof, and a preparation method and application thereof, and solves the technical problems that the ferulic acid and derivatives thereof have unstable bioactivity and are difficult to exert the beneficial effects of whitening, antioxidation, anti-inflammation, acne removal and the like in the field of cosmetics by obtaining the liposome containing the ferulic acid and derivatives thereof with good stability, high encapsulation efficiency and uniform and controllable particle size.

In order to achieve the above object, the present invention provides a method for preparing a liposome containing ferulic acid and derivatives thereof, comprising the following steps:

S10, dispersing ferulic acid and derivatives thereof and an oil-soluble antioxidant into grease to obtain ferulic acid and derivative solutions thereof, and then dissolving phytosterol, phospholipid, glyceride compounds and fatty acyl compounds into an organic solvent to obtain liposome solutions;

S20, mixing the ferulic acid and the derivative solution thereof with the liposome solution, placing the mixture in a sealed container, heating and evaporating the organic solvent under a vacuum condition to obtain a liposome film, then filling nitrogen until the organic solvent is free of residues, continuing to heat until the liposome film falls off, and performing ultrasonic dispersion to form a liposome suspension;

S30, adding glycerol, water and a water-soluble antioxidant into the liposome suspension to form liposome emulsion, and homogenizing to form a liposome coarse material;

and S40, enabling the plastid coarse material body to pass through a composite membrane formed by superposing filter membranes in large, medium and small pore size intervals, and extruding to obtain the liposome containing the ferulic acid and the derivatives thereof.

Optionally, the total weight of the ferulic acid and the derivatives thereof is 1-5 parts, the oil-soluble antioxidant is 0.1-0.5 part, the grease is 1-20 parts, the phytosterol is 1-2 parts, the phospholipid is 1-5 parts, the glyceride compound is 1-5 parts, the fatty acyl compound is 1-10 parts, the sum of the weight of the glycerin and the water is 2-80 parts, and the water-soluble antioxidant is 0.1-0.5 part.

Optionally, the weight ratio of glycerin to water is 1: (3-15).

Optionally, the ferulic acid and the derivatives thereof comprise at least one of ferulic acid, ethylhexyl ferulate and ethyl ferulate and derivatives thereof;

And/or the plant sterol comprises at least one of beta-sitosterol, cholesterol, stigmasterol and ergosterol;

and/or the phospholipid comprises at least one of lecithin, neurophospholipid, phosphatidylserine, phosphatidic acid, hydrogenated lecithin;

and/or the glyceride compound comprises at least one of glycerophospholipids and glycerolipids:

And/or the fatty acyl compound comprises at least one of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylglycerol;

And/or the grease comprises at least one of sunflower seed oil, caprylic/capric triglyceride, ethylhexyl palmitate, nut oil and grape seed oil;

And/or the oil-soluble antioxidant comprises at least one of tocopherol, diethyl hexyl syringylidene malonate, dimethylmethoxy chromanol or a tocopherol derivative;

And/or the water-soluble antioxidant comprises at least one of ascorbic acid and 3-o-ethyl ascorbic acid.

Optionally, in the step of extruding to obtain liposome containing ferulic acid and its derivatives, the extruding temperature is 25-35 ℃, and the extruding pressure is 50-70Mpa.

Optionally, the pore diameter of the large-pore-diameter filter membrane in the composite membrane is 200-400nm, the pore diameter of the medium-pore-diameter filter membrane is 80-200nm, and the pore diameter of the small-pore-diameter filter membrane is less than or equal to 80nm.

Optionally, in the step of homogenizing to form a plastid coarse material, homogenizing treatment is completed by a micro-jet technology, wherein the number of homogenizing cycles of the micro-jet is less than or equal to 4.

Optionally, the filter membrane is a carbon resin filter membrane.

The liposome containing ferulic acid and the derivatives thereof prepared by the preparation method is of a core-shell structure, the core of the liposome comprises ferulic acid and the derivatives thereof, the shell of the liposome is of a liposome bilayer structure, and the liposome bilayer structure comprises phytosterol, phospholipid, glyceride compounds and fatty acyl compounds.

The invention also provides application of the liposome containing the ferulic acid and the derivative thereof in the field of cosmetics, and the liposome-encapsulated ferulic acid and the derivative thereof have more excellent stability and slow release effect, and can effectively exert the beneficial effects of the ferulic acid and the derivative thereof when being applied to the field of cosmetics.

The invention has the beneficial effects that:

The ferulic acid, especially the ferulic acid esterified derivative, has good lipophilicity, can achieve good dispersing effect only by using grease as a solvent without adding a solubilizer, does not have the risks of cytotoxicity caused by the solubilizer, destroying the structure of liposome, affecting the transportation and storage stability and the like, does not generate foam, and does not affect the skin feel during use.

Aiming at the problem that active substances of the ferulic acid and the derivatives thereof are easy to oxidize and inactivate, the invention introduces the oil-soluble antioxidant in the early preparation stage and introduces the water-soluble antioxidant in the later preparation stage, thereby protecting the biological activity of the ferulic acid and the derivatives thereof aiming at each preparation stage which is easy to inactivate the ferulic acid and the derivatives thereof and ensuring that the encapsulated ferulic acid and the derivatives thereof still have higher biological activity.

In the process, the concentration of the plastid coarse material is regulated by mixing glycerol and water, so that the molecular collision rate between liposome double molecular layers can be weakened, the shell structure is prevented from being broken due to the fusion of the encapsulated liposome double molecular layers, the encapsulation stability is improved, and meanwhile, the glycerol can also improve the freeze-thawing stability of the system; in addition, the liposome containing the ferulic acid and the derivative thereof is obtained by superposing and extruding a plurality of layers of films with gradient change of the grain size, and the liposome with smaller grain size, uniform grain size and high dispersivity can be obtained without replacing the film layers for a plurality of times.

The liposome containing the ferulic acid and the derivative thereof has excellent stability and long-acting slow release effect, and the liposome is used for conveying the ferulic acid and the derivative thereof from the epidermis of the skin to the inside of the skin, thereby being more beneficial to the effects of whitening, antioxidation, anti-inflammation, acne removal and the like of the ferulic acid and the derivative thereof.

Drawings

For a clearer description of embodiments of the invention or of solutions in the prior art, the following brief description of the drawings is given for the purpose of illustrating the embodiments or the solutions in the prior art, it being obvious that the drawings in the following description are only some embodiments of the invention, and that other drawings may be obtained from the structures shown in these drawings without the need for inventive effort for a person skilled in the art.

FIG. 1 is a schematic flow chart of a method for preparing a liposome containing ferulic acid and derivatives thereof according to the present invention.

FIG. 2 is an SEM image of the distribution of the liposome containing ferulic acid and its derivatives in solution according to example 1 of the present invention.

FIG. 3 is an SEM image of gel products made of liposome containing ferulic acid and its derivatives obtained in example 1 of the present invention.

The achievement of the objects, functional features and advantages of the present invention will be further described with reference to the accompanying drawings, in conjunction with the embodiments.

Detailed Description

It should be understood that the specific embodiments described herein are for purposes of illustration only and are not intended to limit the scope of the invention.

The following description of the embodiments of the present invention will be made clearly and fully with reference to the accompanying drawings, in which it is evident that the embodiments described are only some, but not all embodiments of the invention. All other embodiments, which can be made by those skilled in the art based on the embodiments of the invention without making any inventive effort, are intended to be within the scope of the invention.

The description as it relates to "first", "second", etc. in the present invention is for descriptive purposes only and is not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include at least one such feature. In addition, the technical solutions of the embodiments may be combined with each other, but it is necessary to base that the technical solutions can be realized by those skilled in the art, and when the technical solutions are contradictory or cannot be realized, the combination of the technical solutions should be considered to be absent and not within the scope of protection claimed in the present invention.

The invention provides a preparation method of liposome containing ferulic acid and derivatives thereof, referring to figure 1, the preparation method of liposome containing ferulic acid and derivatives thereof comprises the following steps:

s20, mixing the ferulic acid and the derivative solution thereof with the liposome solution, placing the mixture in a sealed container, heating and evaporating the organic solvent under a vacuum condition to obtain a liposome film, then filling nitrogen until the organic solvent is free of residues, continuing heating until the liposome film falls off, and performing ultrasonic dispersion to form a liposome suspension;

The ferulic acid and the derivative thereof comprise at least one of ferulic acid, ethylhexyl ferulate and ethyl ferulate, and preferably at least one of ethylhexyl ferulate and ethyl ferulate.

In some embodiments, the ferulic acid and derivatives thereof may be ferulic acid.

In some embodiments, the ferulic acid and derivatives thereof may be ethylhexyl ferulate.

In some embodiments, the ferulic acid and derivatives thereof may be ethyl ferulate.

In some embodiments, the ferulic acid and derivatives thereof may be ferulic acid and ethylhexyl ferulate.

In some embodiments, ferulic acid and its derivatives may be ferulic acid and ethyl ferulate.

In some embodiments, the ferulic acid and its derivatives may be ferulic acid, ethylhexyl ferulate, and ethyl ferulate.

In some embodiments, ferulic acid and its derivatives may be ethylhexyl ferulate and ethyl ferulate.

The ferulic acid, especially the ferulic acid esterified derivative, has excellent lipophilicity, is easy to dissolve in grease, adopts the grease as a solvent, can achieve good dispersion effect and stability without adding a solubilizer, does not have the risks of cytotoxicity caused by the solubilizer, destroying the structure of liposome, affecting the transportation and storage stability of the liposome and the like, does not generate foam, and does not affect the skin feel during use.

The present invention is not limited to the type of the oil or fat, and the oil or fat is preferably a polar oil or fat, and may be at least one of sunflower seed oil, caprylic/capric triglyceride, ethylhexyl palmitate, nut oil, and grape seed oil, for example. The grease is used as a solvent, so that ferulic acid and derivatives thereof can be uniformly dispersed, and good skin-friendly property can be given to the product.

In order to ensure that better biological stability of the ferulic acid and the derivatives thereof is maintained after being coated, antioxidants are introduced in the process of dispersing the ferulic acid and the derivatives thereof into the grease in the step S10 and in the process of adding glycerol and water into the liposome suspension in the step S30.

Wherein the system of step S10 is predominantly fat-soluble, thus incorporating an oil-soluble antioxidant, preferably comprising at least one of tocopherol, diethylhexyl syringylidene malonate, dimethylmethoxy chromanol or a tocopherol derivative; in step S30, water and glycerin are added, and the water-soluble antioxidants are well dissolved, so that the water-soluble antioxidants including at least one of ascorbic acid and 3-o-ethyl ascorbic acid are used. The staged addition of the antioxidant can ensure the stability of the ferulic acid and the derivatives thereof in the preparation process and ensure that the encapsulated ferulic acid and the derivatives thereof still have higher biological activity.

Liposomes are the main components constituting biological membranes, participate in signaling, regulate the signaling processes of cell growth, differentiation, aging, programmed death, etc., and provide energy, participate in body growth, and maintain vital activities. The shell part of the liposome containing the ferulic acid and the derivative thereof is a liposome bilayer structure, comprises a phytosterol, a phospholipid, a glyceride compound and a fatty acyl compound, constructs a complete and compact shell oil film structure suitable for the ferulic acid and the derivative thereof, and aims to improve the stability, the dispersibility and the encapsulation rate of the liposome containing the ferulic acid and the derivative thereof.

In some embodiments, the phytosterol comprises at least one of beta-sitosterol, cholesterol, stigmasterol and ergosterol, and the use of said phytosterol can strengthen the bilayer membrane structure of the liposome, reduce membrane flow and reduce leakage rate.

In some embodiments, the phospholipid comprises at least one of lecithin, neurophospholipid, phosphatidylserine, phosphatidic acid, hydrogenated lecithin.

In some embodiments, the glyceride compound comprises at least one of a triglyceride, a triglyceride oil; the glyceride compound can be used together with other lipids, so that the embedding effect on the ferulic acid and the derivatives thereof can be improved, the problems that the ferulic acid and the derivatives thereof are not easy to permeate into the skin horny layer and are unstable and easy to inactivate can be solved, and the long-acting slow release effect can be achieved.

In some embodiments, the fatty acyl compound includes at least one of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylglycerol.

The invention is not limited to the parts by weight of all the above components, in some embodiments, the total weight of the ferulic acid and the derivatives thereof is 1 to 5 parts by weight, the oil-soluble antioxidant is 0.1 to 0.5 part by weight, the grease is 1 to 20 parts by weight, the phytosterol is 1 to 2 parts by weight, the phospholipid is 1 to 5 parts by weight, the glyceride compound is 1 to 5 parts by weight, the fatty acyl compound is 1 to 10 parts by weight, the sum of the glycerin and the water is 2 to 80 parts by weight, the water-soluble antioxidant is 0.1 to 0.5 part by weight, and further, the weight ratio of the glycerin to the water is 1: (3-15). In the proportion range, the liposome containing the ferulic acid and the derivatives thereof with high encapsulation efficiency and stability can be obtained, and the problems that the drug leakage is caused by the fact that the dosage of the ferulic acid and the derivatives thereof is too large and exceeds the quantity which can be carried by the liposome, or the dosage of the ferulic acid and the derivatives thereof is too small, the coated drug quantity is too small, the advantage of the liposome serving as a drug delivery system is lost and the like can be avoided.

The liposome suspension in the step S20 is in a microemulsion state, the concentration of active substances is high, and because the higher the concentration of the active substances is, the faster the collision frequency among molecules is, so that the bilayer membranes of the liposome are easy to dissolve and the encapsulation structure of the liposome is broken, therefore, the invention mixes water and glycerol, adjusts the viscosity of a system, reduces the concentration of the active substances, reduces the collision among molecules, protects the shell structure of the liposome, improves the encapsulation rate, and further ensures the bioactivity of ferulic acid and derivatives thereof.

Because the liposome suspension is in a microemulsion state, the emulsion can be frozen and melted when meeting low-temperature conditions, the stability of the emulsion can be influenced, the emulsion is coagulated when the emulsion is heavy, the apparent viscosity of the emulsion is increased when the emulsion is light, and the glycerol added in the invention also plays a role in improving the freeze-thawing stability of a system.

Further, more specifically, in step S10:

the organic solvent used for dissolving the phytosterol, the phospholipid, the glyceride compound and the fatty acyl compound is preferably methylene dichloride, because methylene dichloride has better solubility and lower boiling point, and can be removed by a heating mode such as rotary evaporation and the like.

In some embodiments, after the phytosterol, phospholipid, glyceride compound, fatty acyl compound is added into the organic solvent, the dissolution is accelerated by water bath ultrasonic method at 30-40 ℃.

In step S20:

Preferably, the organic solvent is evaporated under vacuum by heating to 40-45℃for 1-2h. Under the conditions of temperature and time, not only can the organic solvent be removed by evaporation, but also the biological activity and stability of the ferulic acid and the derivatives thereof can be ensured not to be destroyed.

In some embodiments, vacuum pumping is performed for 20-40min to form a uniform liposome film on the inner wall of the sealed container, and then nitrogen is filled until no organic solvent remains, so that the organic solvent is prevented from damaging human body or destroying liposome shell structure.

In some embodiments, the heating is continued by means of a water bath spin-steaming, preferably at a temperature of 35-40 ℃, and preferably at 60rpm, until the liposome membrane is shed.

In some embodiments, the liposome suspension is formed by ultrasonic dispersion under ice bath conditions, so that the biological activity and stability of the ferulic acid and the derivatives thereof can be ensured not to be destroyed.

In step S30:

The homogenization temperature is preferably 30-40 ℃, preferably low-temperature homogenization, and is also used for ensuring the bioactivity and encapsulation effect of the ferulic acid and the derivative thereof and the color forming quality of the product, and if the encapsulation rate is low, the ferulic acid and the derivative thereof are free outside the liposome structure and are easy to oxidize and change color under the conditions of high temperature and light.

The invention is not limited to the homogenization mode, preferably, homogenization is completed by a micro-jet technology, liposome coarse bodies with smaller particle size and uniform dispersion can be obtained, and the preparation time is shortened. Further preferably, the number of homogenization cycles of the microfluidics is less than or equal to 4, when the number of homogenization cycles is less than or equal to 4, a liposome coarse body with smaller average particle size can be obtained, and when the number of homogenization cycles is more than 4, the average particle size of the liposome coarse body is increased instead, because the increase of the number of homogenization cycles easily causes the local temperature of the liposome emulsion to be increased, the aggregation phenomenon of liquid drops is caused, and then the average particle size of the liposome coarse body is increased, and the dispersibility of the system is also affected.

Further, the shear flow rate unit of the micro-jet is preferably 200 to 500ml/min.

In step S40:

and (3) superposing the plastid coarse material body by an artificial membrane with three pore diameters of large, medium and small to obtain the composite membrane, and extruding to obtain the liposome containing the ferulic acid and the derivatives thereof.

In view of the difficulty in extrusion to obtain liposome with uniform particle size, the invention selects a composite membrane formed by overlapping artificial membranes with three pore size intervals of large, medium and small as an extrusion filter membrane, and the particle size of the liposome can be controlled according to the requirement.

In some embodiments, the liposome coarse material is placed in a customizable high-pressure cavity, a sample is directly conveyed through pneumatic gas in a pressure mode through a composite membrane formed by overlapping artificial membranes of three pore size intervals, and pressure stability is controlled, so that all the liposome coarse material is extruded by the same extrusion force, and the liposome containing ferulic acid and derivatives thereof with specified particle sizes is obtained.

Preferably, when the multi-layer membrane is combined for superposition, the pore diameter of the large-pore-diameter filter membrane ranges from 200nm to 400nm, the pore diameter of the small-pore-diameter filter membrane is less than or equal to 80nm, and the filter membrane with the transitional pore diameter, preferably the pore diameter of 80-200nm, can be properly selected in the middle. For example, 400nm, 100nm and 80nm may be employed; 300nm, 100nm and 60nm;200nm, 100nm and 50nm;350nm, 150nm and 80nm;250nm, 90nm and 60nm; the combination of the filters was performed with a gradient of 200nm, 80nm, 50nm, etc. The extrusion process is completed through the combination of the filter membranes with three particle sizes with gradient change, the filter membranes do not need to be replaced in the middle, and the liposome containing the ferulic acid and the derivatives thereof with uniform particle sizes can be obtained.

The invention is not limited to the type of artificial membrane, and in some embodiments, the artificial membrane is a carbon resin filter membrane.

The extrusion temperature in this step is preferably 25 to 35℃and the extrusion pressure is preferably 50 to 70MPa. The preferable extrusion temperature and extrusion pressure can ensure the structural integrity of the liposome, improve the encapsulation efficiency and ensure that the biological activity of the ferulic acid and the derivatives thereof is not destroyed.

The particle size of the liposome containing ferulic acid and the derivative thereof prepared by the preparation method is less than or equal to 80nm, and the embedding rate is more than 85%. The liposome containing the ferulic acid and the derivative thereof has excellent stability, dispersibility and long-acting slow release effect, and the liposome is used for conveying the ferulic acid and the derivative thereof from the epidermis of the skin to the inside of the skin, thereby being more beneficial to playing the beneficial effects of whitening, antioxidation, anti-inflammation, acne removal and the like of the ferulic acid and the derivative thereof.

The technical scheme of the present invention will be further described in detail with reference to the following specific examples, which are to be construed as merely illustrative, and not limitative of the remainder of the disclosure.

Example 1

Referring to fig. 1, fig. 1 is a schematic flow chart of an embodiment 1 of a method for preparing a liposome containing ferulic acid and derivatives thereof according to the present invention, which comprises the following steps:

S10, dispersing 2 parts of ethylhexyl ferulate, 2 parts of ferulic acid, 1 part of ethyl ferulate and 0.4 part of tocopherol into 5 parts of sunflower seed oil and 5 parts of caprylic/capric triglyceride, carrying out water bath ultrasonic treatment at 30 ℃ until the mixture is uniformly mixed to obtain ferulic acid and a derivative solution thereof, and then adding 1 part of beta-sitosterol, 1 part of lecithin, 1 part of neurophospholipid, 1 part of hydrogenated lecithin, 5 parts of glycerophospholipid, 1 part of phosphatidylcholine and 4 parts of phosphatidylglycerol into a penicillin bottle filled with dichloromethane, and carrying out water bath ultrasonic treatment at 30 ℃ until the mixture is completely dissolved to obtain a liposome solution.

S20, transferring a liposome solution into a round-bottom flask, slowly adding the ferulic acid and the derivative solution thereof into the round-bottom flask through an organic injection method, adjusting the water bath temperature to 40 ℃, removing dichloromethane through rotary evaporation for 2h under a vacuumizing state, continuously vacuumizing for 30min to form a layer of uniform liposome film on the inner wall of the round-bottom flask, then taking down the round-bottom flask, rapidly filling pure nitrogen for 1min to ensure that dichloromethane is free from residues, and then rotating in a water bath at a rotating speed of 60rpm under normal pressure and at 35 ℃ to enable the liposome film to fall off, and then performing ice bath ultrasonic dispersion on the liposome film to form a liposome suspension;

S30, adding 10 parts of glycerol, 30 parts of deionized water and 0.3 part of 3-o-ethyl ascorbic acid into the liposome suspension, uniformly mixing to form liposome emulsion, homogenizing the liposome emulsion for 1h by a micro-jet homogenizer at a shearing flow of 300ml/min and a homogenizing temperature of 30 ℃, and carrying out homogenizing circulation for 4 times to obtain a liposome coarse material;

S40, the plastid coarse material is placed in a high-pressure cavity, the plastid coarse material is directly pressed and conveyed through constant air pressure of 60Mpa, and a composite membrane formed by superposing three layers of carbon resin filter membranes with pore diameters of 400nm, 200nm and 80nm is extruded at 25 ℃ to obtain the liposome containing the ferulic acid and the derivatives thereof.

Example 2

Referring to fig. 1, fig. 1 is a schematic flow chart of an embodiment 2 of a method for preparing a liposome containing ferulic acid and its derivatives according to the present invention, which comprises the following steps:

S10, dispersing 1 part of ferulic acid, 1 part of ethyl ferulate and 0.5 part of diethyl hexyl syringylidene malonate into 20 parts of caprylic/capric triglyceride, performing water bath ultrasonic treatment at 35 ℃ until the mixture is uniform to obtain ferulic acid and derivative solution thereof, adding 1 part of cholesterol, 1 part of beta-sitosterol, 3 parts of hydrogenated lecithin, 2 parts of lecithin, 5 parts of triglyceride, 5 parts of phosphatidylcholine and 5 parts of phosphatidylethanolamine into a penicillin bottle filled with dichloromethane, and performing water bath ultrasonic treatment at 30 ℃ until the solution is completely dissolved to obtain liposome solution;

S20, transferring a liposome solution into a round-bottom flask, slowly adding the ferulic acid and the derivative solution thereof into the round-bottom flask through an organic injection method, adjusting the water bath temperature to be 42 ℃, removing dichloromethane through rotary evaporation for 1h under a vacuumizing state, continuously vacuumizing for 40min to enable the inner wall of the round-bottom flask to form a layer of uniform liposome film, then taking down the round-bottom flask, rapidly filling pure nitrogen for 1min to enable dichloromethane to have no residue, and then rotating in a water bath at a rotating speed of 60rpm under normal pressure and 38 ℃ to enable the liposome film to fall off, and then performing ice bath ultrasonic dispersion on the liposome film to form a liposome suspension;

S30, adding 1 part of glycerol, 15 parts of deionized water, 0.1 part of 3-o-ethyl ascorbic acid and 0.1 part of ascorbic acid into the liposome suspension, uniformly mixing to form liposome emulsion, homogenizing the liposome emulsion for 2 hours by a micro-jet homogenizer at a shearing flow rate of 400ml/min and a homogenizing temperature of 40 ℃, and carrying out homogenizing circulation for 3 times to obtain a liposome coarse material;

S40, the plastid coarse material is placed in a high-pressure cavity, the plastid coarse material is directly pressed and conveyed through constant air pressure of 70Mpa, and the plastid coarse material is extruded at 35 ℃ through a composite membrane formed by superposing three layers of carbon resin filter membranes with apertures of 400nm, 200nm and 80nm respectively.

Example 3

Referring to fig. 1, fig. 1 is a schematic flow chart of an embodiment 3 of a method for preparing a liposome containing ferulic acid and derivatives thereof according to the present invention, which comprises the following steps:

S10, dispersing 0.5 part of ethylhexyl ferulate, 0.5 part of ferulic acid and 0.2 part of tocopherol into 1 part of ethylhexyl palmitate, performing water bath ultrasonic treatment at 40 ℃ until the mixture is uniformly mixed to obtain a solution of ethylhexyl ferulate, adding 1.5 parts of ergosterol, 1 part of hydrogenated lecithin, 2 parts of lecithin, 3 parts of triglyceride and 1 part of phosphatidylglycerol into a penicillin bottle filled with dichloromethane, and performing water bath ultrasonic treatment at 40 ℃ for 10 minutes until the solution is completely dissolved to obtain a liposome solution;

S20, transferring the liposome solution into a round-bottom flask, slowly adding the ferulic acid and the derivative solution thereof into the round-bottom flask through an organic injection method, adjusting the water bath temperature to 45 ℃, removing dichloromethane through rotary evaporation for 1.5h under a vacuumizing state, continuously vacuumizing for 20min to enable the inner wall of the round-bottom flask to form a layer of uniform liposome film, then taking down the round-bottom flask, rapidly filling pure nitrogen for 1min to enable dichloromethane to be free of residues, and then rotating in a water bath at a rotating speed of 60rpm under normal pressure and at 40 ℃ to enable the liposome film to fall off, and then performing ice bath ultrasonic dispersion on the liposome film to form liposome suspension;

S30, adding 0.5 part of glycerol, 1.5 parts of deionized water and 0.5 part of ascorbic acid into the liposome suspension, uniformly mixing to form liposome emulsion, homogenizing the liposome emulsion for 1h by a micro-jet homogenizer at a shearing flow rate of 500ml/min and a homogenizing temperature of 35 ℃, and carrying out homogenizing circulation for 2 times to obtain a plastid coarse material;

S40, the plastid coarse material is placed in a high-pressure cavity, the plastid coarse material is directly pressed and conveyed through constant air pressure of 55Mpa, and a composite membrane formed by overlapping three layers of carbon resin filter membranes with the pore diameters of 200nm, 100nm and 50nm is extruded at the temperature of 30 ℃ to obtain the product.

Example 4

Referring to fig. 1, fig. 1 is a schematic flow chart of an embodiment 4 of a method for preparing a liposome containing ferulic acid and derivatives thereof according to the present invention, which comprises the following steps:

S10, dispersing 1 part of ferulic acid, 1 part of ethylhexyl ferulate and 0.1 part of dimethyl methoxy chromanol into 8 parts of grape seed oil according to parts by weight, carrying out water bath ultrasonic treatment at 35 ℃ until the mixture is uniformly mixed to obtain a ferulic acid solution, adding 2 parts of beta-sitosterol, 2 parts of lecithin, 1 part of glycerophosphate, 4 parts of phosphatidylcholine and 4 parts of phosphatidylinositol into a penicillin bottle filled with dichloromethane, and carrying out water bath ultrasonic treatment at 35 ℃ for 10 minutes until the ferulic acid solution is completely dissolved to obtain a liposome solution;

s20, transferring a liposome solution into a round-bottom flask, slowly adding the ferulic acid and the derivative solution thereof into the round-bottom flask through an organic injection method, adjusting the water bath temperature to 44 ℃, removing dichloromethane through rotary evaporation for 1h under a vacuumizing state, continuously performing vacuum suction for 30min to enable the inner wall of the round-bottom flask to form a layer of uniform liposome film, then taking down the round-bottom flask, rapidly filling pure nitrogen for 1min to enable dichloromethane to have no residue, and then rotating in a water bath at a rotating speed of 60rpm under normal pressure and at 35 ℃ to enable the liposome film to fall off, and then performing ice bath ultrasonic dispersion on the liposome film to form a liposome suspension;

S30, adding 8 parts of glycerol, 72 parts of deionized water and 0.4 part of 3-o-ethyl ascorbic acid into the liposome suspension, uniformly mixing to form liposome emulsion, homogenizing the liposome emulsion for 1.5h by a micro-jet homogenizer at a shearing flow of 200ml/min and a homogenizing temperature of 35 ℃, and carrying out homogenizing circulation for 1 time to obtain a plastid coarse material;

S40, the plastid coarse material is placed in a high-pressure cavity, the plastid coarse material is directly pressed and conveyed through constant air pressure of 65Mpa, and the plastid coarse material is extruded at 25 ℃ through a composite membrane formed by superposing three layers of carbon resin filter membranes with the pore diameters of 200nm, 100nm and 50nm respectively.

Example 5

Referring to fig. 1, fig. 1 is a schematic flow chart of an embodiment 5 of a method for preparing a liposome containing ferulic acid and derivatives thereof according to the present invention, which comprises the following steps:

s10, dispersing 5 parts of ethyl ferulate, 0.1 part of tocopherol and 0.2 part of diethyl hexyl syringylidene malonate into 5 parts of nut oil and 5 parts of grape seed oil, carrying out water bath ultrasonic treatment at 35 ℃ until the mixture is uniform to obtain an ethyl ferulate solution, adding 1 part of stigmasterol, 1 part of hydrogenated lecithin, 2 parts of triglyceride, 2 parts of phosphatidylcholine and 1 part of phosphatidylethanolamine into a penicillin bottle filled with dichloromethane, and carrying out water bath ultrasonic treatment at 35 ℃ for 10min until the mixture is completely dissolved to obtain a liposome solution;

S20, transferring a liposome solution into a round-bottom flask, slowly adding the ethyl ferulate solution into the round-bottom flask through an organic injection method, adjusting the water bath temperature to 40 ℃, removing dichloromethane through rotary evaporation for 2h under a vacuumizing state, continuously vacuumizing for 20min to form a layer of uniform liposome film on the inner wall of the round-bottom flask, then taking down the round-bottom flask, quickly filling pure nitrogen for 1min to ensure that dichloromethane is free from residues, and then rotating in a water bath at a rotating speed of 60rpm under the conditions of normal pressure and 40 ℃ to enable the liposome film to fall off, and then performing ice bath ultrasonic dispersion on the liposome film to form a liposome suspension;

S30, adding 2 parts of glycerol, 24 parts of deionized water and 0.1 part of ascorbic acid into the liposome suspension, uniformly mixing to form liposome emulsion, homogenizing the liposome emulsion for 2 hours by a micro-jet homogenizer at a shearing flow of 300ml/min and a homogenizing temperature of 35 ℃, and carrying out homogenizing circulation for 4 times to obtain a liposome coarse material;

S40, the plastid coarse material is placed in a high-pressure cavity, the plastid coarse material is directly pressed and conveyed through constant air pressure of 50Mpa, and the plastid coarse material is extruded at 35 ℃ through a composite membrane formed by superposing three layers of carbon resin filter membranes with the pore diameters of 300nm, 100nm and 60nm respectively.

Comparative example 1

Comparative example 1 was prepared in the same manner as in example 1, except that no antioxidant was added in step S10 and step S30.

Comparative example 2

Comparative example 2 was prepared in the same manner as in example 1, except that the glycerin and deionized water were 20 parts by weight and 20 parts by weight, respectively, in step S30.

Comparative example 3

Comparative example 3 was prepared in the same manner as in example 1, except that in step S30, the weight parts of glycerin and deionized water were 2 parts and 38 parts, respectively.

Comparative example 4

Comparative example 4 was the same as in example 1 in terms of the preparation method, except that in step S40, the plastid coarse material was passed through a layer of polycarbonate resin filter membrane having a particle size of 80nm.

Comparative example 5

Comparative example 5 was the same as example 1 in the feed, except that in step S40, the homogenization was repeated 5 times.

Comparative example 6

Comparative example 6 was the same as example 1 in the procedure of preparation except that no nitrogen was added to dichloromethane to leave no residue after the liposome membrane was obtained by vacuum suction in step S20.

Performance testing

1. Encapsulation efficiency: the products of the examples and comparative examples were subjected to a post-separation assay using a gel chromatography column, and the encapsulation efficiency was calculated as follows:

Encapsulation efficiency (%) = (amount of encapsulated drug in system/total amount of encapsulated and unencapsulated drug in system) ×100%.

2. PDI index: the PDI index of the products of the examples and comparative examples was determined.

3. 90 Day stability test: the products of examples and comparative examples were prepared as 5% by mass aqueous solutions and left for 90 days to see if any oil had evolved.

4. Freeze thawing resistance test: the products of the examples and the comparative examples were subjected to 5 freeze-thawing cycles (23.+ -. 2) C for 16h in water, then frozen at-25 ℃ for 8h, finally placed at normal temperature, and after melting with ice, subjected to the next cycle. After 5 cycles, it was observed whether the system could be restored to its original state or delamination occurred.

The results of the above tests 1-4 are shown in Table 1.

5. Morphology observation: characterization of the mixed solution of the liposome containing ferulic acid and the derivatives thereof obtained in example 1 by using an electron scanning microscope is shown in fig. 2; the liposome obtained in example 1 and containing ferulic acid and its derivatives were prepared into gel products, and the results were characterized by electron scanning microscopy and are shown in FIG. 3.

TABLE 1 comparison of Properties of the products obtained in examples 1-5 and comparative examples 1-6

Test item	Encapsulation efficiency%	PDI	Stability for 90 days	Freeze thawing resistance
					Example 1	87	0.24	Stabilization	Stabilization
Example 2	89	0.18	Stabilization	Stabilization
					Example 3	86	0.15	Stabilization	Stabilization
Example 4	86	0.16	Stabilization	Stabilization
					Example 5	90	0.21	Stabilization	Stabilization
Comparative example 1	75	0.42	Has oily matter separated out	Unable to recover the original state, delamination of the solution
					Comparative example 2	75	0.44	Has oily matter separated out	Unable to recover the original state, delamination of the solution
Comparative example 3	73	0.38	Has oily matter separated out	Unable to recover the original state, delamination of the solution
					Comparative example 4	68	0.40	Has oily matter separated out	Unable to recover the original state, delamination of the solution
Comparative example 5	63	0.42	Has oily matter separated out	Unable to recover the original state, delamination of the solution
					Comparative example 6	75	0.52	Has oily matter separated out	Unable to recover the original state, delamination of the solution

As can be seen from fig. 2: the liposome containing ferulic acid and its derivatives has uniform distribution and uniform particle size.

As can be seen from fig. 3: the gel product prepared from liposome containing ferulic acid and its derivatives has uniform texture, no ion, and good stability.

As can be seen from table 1:

The experimental data can be obtained, the encapsulation rate of the embodiments 1 to 5 is higher and reaches more than 85%, the PDI value is smaller, the dispersibility is better, the stability is good, the substance precipitation and layering phenomenon are not easy to occur, the materials can be quickly recovered through a freeze thawing resistance test, the dispersion is uniform, the layering and coagulation turbidity phenomenon are not caused, and the anti-freezing agent has higher freeze thawing resistance.

In comparative example 1, no antioxidant is used, the biological activity of ferulic acid and the derivatives thereof is affected, the dispersion performance of the product is also affected, the product is placed for 90 days, oily matters are separated out, the product cannot be restored to the original state after a freeze-thawing test, layering occurs, and the stability is not ideal.

Comparative example 2 added too much glycerol and comparative example 3 added too little glycerol was detrimental to both storage stability and freeze-thaw stability of the product, and glycerol could not be properly proportioned with water to bring the system to a stable state, which in turn resulted in lower encapsulation efficiency.

Comparative example 4 using a layer of a polycarbonate resin filter membrane with a particle size of 80nm for extrusion may cause the shell structure of the liposome containing ferulic acid and its derivatives to be broken, resulting in lower encapsulation efficiency, high encapsulation efficiency without extrusion through a stepped three-layer filter membrane, and poor encapsulation efficiency also results in precipitation of oily substances in a 90-day stability test, failure to recover the original state after freeze-thaw stability test, and delamination.

The homogenization frequency of comparative example 5 reaches 5 times, the local temperature in the system is increased, so that the aggregation phenomenon of liquid drops in the system is caused, the dispersibility of the product is not improved, the homogenization frequency is too high, the shell of the liposome is possibly broken, the encapsulation rate is low, oily substances are separated out of the product in a 90-day stability experiment, the product cannot be restored to the original state after a freeze-thawing stability test, and layering occurs.

Comparative example 6 was not filled with nitrogen, and residual dichloromethane was easy, and not only the skin, digestive tract and respiratory tract entered the human body, produced the result of corrosion and poisoning, but also the liposome shell structure was broken, the encapsulation efficiency of the product was lower, and the stability was poor.

The foregoing description is only of the preferred embodiments of the present invention, and is not intended to limit the scope of the invention, but rather is intended to cover any equivalents of the structures or equivalent processes disclosed herein or in the alternative, which may be employed directly or indirectly in other related arts.

Claims

1. A method for preparing a liposome containing ferulic acid and derivatives thereof, comprising the steps of:

S20, mixing the ferulic acid and the derivative solution thereof with the liposome solution, placing the mixture in a sealed container, heating and evaporating the organic solvent under a vacuum condition, then performing vacuum suction to obtain a liposome film, then charging nitrogen until the organic solvent is free from residue, continuously heating until the liposome film falls off, and performing ultrasonic dispersion to form a liposome suspension;

S40, enabling the plastid coarse material body to pass through a composite membrane formed by superposing filter membranes in large, medium and small pore size intervals, and extruding to obtain a liposome containing ferulic acid and derivatives thereof;

The weight ratio of the glycerol to the water is 1: (3-15);

1-5 parts of ferulic acid and derivatives thereof, 0.1-0.5 part of oil-soluble antioxidant, 1-20 parts of grease, 1-2 parts of phytosterol, 1-5 parts of phospholipid, 1-5 parts of glyceride compound, 1-10 parts of fatty acyl compound, 2-80 parts of sum of glycerol and water and 0.1-0.5 part of water-soluble antioxidant;

The ferulic acid and the derivatives thereof comprise at least one of ferulic acid, ethylhexyl ferulate and ethyl ferulate;

the plant sterol comprises at least one of beta-sitosterol, cholesterol, stigmasterol and ergosterol;

The phospholipid comprises at least one of lecithin, neurophospholipid, phosphatidylserine, phosphatidic acid and hydrogenated lecithin;

The glyceride compound comprises at least one of glycerophospholipids and glycerolipids:

The fatty acyl compound comprises at least one of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol and phosphatidylglycerol;

The oil comprises at least one of sunflower seed oil, caprylic/capric triglyceride, ethylhexyl palmitate, nut oil and grape seed oil;

the oil-soluble antioxidant comprises at least one of tocopherol, diethyl hexyl syringylidene malonate, dimethyl methoxy chromanol or a tocopherol derivative;

The water-soluble antioxidant comprises at least one of ascorbic acid and 3-o-ethyl ascorbic acid;

in the step of homogenizing to form a plastid coarse material body, homogenizing treatment is completed by a micro-jet technology, and the number of homogenizing cycles of the micro-jet is less than or equal to 4.

2. The method for preparing a liposome containing ferulic acid and its derivatives as claimed in claim 1, wherein in the step of extruding to obtain the liposome containing ferulic acid and its derivatives, the extrusion temperature is 25-35 ℃, and the extrusion pressure is 50-70Mpa.

3. The method for preparing the liposome containing the ferulic acid and the derivatives thereof according to claim 1, wherein the pore size of the large pore size filter membrane in the composite membrane is 200-400nm, the pore size of the medium pore size filter membrane is 80-200nm, and the pore size of the small pore size filter membrane is less than or equal to 80nm.

4. The method for preparing a liposome containing ferulic acid and its derivatives as claimed in claim 1, wherein the filter membrane is a carbon resin filter membrane.

5. The liposome containing ferulic acid and derivatives thereof prepared by the preparation method according to any one of claims 1 to 4, wherein the liposome containing ferulic acid and derivatives thereof has a core-shell structure, the core of the liposome containing ferulic acid and derivatives thereof has a liposome bilayer structure, and the liposome bilayer structure contains phytosterol, phospholipid, glyceride compounds and fatty acyl compounds.

6. Use of the liposome containing ferulic acid and its derivatives according to claim 5 in the cosmetic field.