CN115227589A

CN115227589A - Stabilized resveratrol inclusion and preparation method thereof

Info

Publication number: CN115227589A
Application number: CN202211078679.7A
Authority: CN
Inventors: 常宽; 贾宝通; 许虎君; 高海燕; 林良良; 沈洁
Original assignee: Jiangnan University
Current assignee: Jiangnan University
Priority date: 2022-09-05
Filing date: 2022-09-05
Publication date: 2022-10-25
Anticipated expiration: 2042-09-05
Also published as: CN115227589B

Abstract

The invention discloses a stabilized resveratrol inclusion and a preparation method thereof, belonging to the technical field of daily chemical industry. The preparation method of the resveratrol inclusion body comprises the following steps: (1) Mixing and stirring phospholipid and auxiliary emulsifier to obtain emulsifier composition; (2) And (2) stirring and mixing the emulsifier composition prepared in the step (1) and the oil phase, adding the water phase, and continuously stirring to obtain the stabilized resveratrol inclusion. The inclusion prepared by the invention has excellent thermodynamic stability, precipitation and demulsification of the resveratrol can not be caused after long-term storage, the inclusion can effectively inhibit degradation and discoloration of the resveratrol, the preparation process is simple and easy to implement, and the requirement on equipment is low.

Description

Stabilized resveratrol inclusion and preparation method thereof

Technical Field

The invention relates to the technical field of daily chemical industry, in particular to a stabilized resveratrol inclusion body and a preparation method thereof.

Background

Resveratrol, with the scientific name of 3,4', 5-trihydroxy-1,2-diphenylethylene, is a natural non-flavonoid polyphenol compound. Resveratrol is a natural antioxidant, and has excellent biological activities of resisting aging, resisting bacteria and cancer and preventing cardiovascular diseases, so that resveratrol is widely concerned and applied in the fields of daily chemicals, foods and medicines.

The photo-thermal stability and solubility of resveratrol have long been a major problem affecting the application of its products. Resveratrol is insoluble in water and poor in oil solubility, can be dissolved in only a few oils, and is easy to separate out after being dissolved in most oils. In addition, the resveratrol is easy to react with oxygen under the conditions of illumination and heating, and is degraded and browned. Therefore, if the resveratrol is directly applied to daily chemical and medical products as an oil phase active substance, the problems of precipitation, oxidative degradation, product discoloration, unstable formula and the like of the resveratrol can be caused in the storage and transportation processes of the products.

The improvement of the solubility, the oxidation resistance and the stability of the resveratrol which is an active ingredient of the cosmetics are the problems worth of research in daily chemical research and development, and have important practical significance. At present, in order to solve the problems, an antioxidant is generally added or a coating technology is adopted to prevent the raw materials from oxidative degradation. The addition of antioxidants can present challenges to formulation compatibility, and the excessive use of antioxidants in daily use can greatly increase the risk of canceration of the human body and cause some irreversible physiological damage to the body. The packaging technology of resveratrol mainly comprises liposome packaging, and the preparation method of liposome is complicated, involves complicated equipment and organic solvent, and is difficult to produce and apply in large scale.

Disclosure of Invention

Aiming at the defects of the prior art, the invention provides a stabilized resveratrol inclusion body and a preparation method thereof. According to the invention, the cosmetic raw materials are wrapped by a micro-nano emulsion wrapping technology, so that the stability is enhanced, the problems of oxidative degradation and discoloration in the storage process of the cosmetic raw materials are inhibited, the prepared inclusion has excellent thermodynamic stability, precipitation and emulsion breaking of resveratrol cannot be caused even if the inclusion is stored for a long time, the inclusion can effectively inhibit degradation and discoloration of resveratrol, the preparation process is simple and easy to implement, and the requirement on equipment is low.

The technical scheme of the invention is as follows:

a preparation method of a stabilized resveratrol inclusion body comprises the following steps:

(1) Mixing and stirring phospholipid and auxiliary emulsifier to obtain emulsifier composition;

(2) And (2) stirring and mixing the emulsifier composition prepared in the step (1) and the oil phase, adding the water phase, and continuously stirring to obtain the stabilized resveratrol inclusion.

Further, in the step (1), the phospholipid is one or more of soybean phospholipid, lecithin, cephalin and cardiolipin.

Further, in the step (1), the co-emulsifier is composed of lower alcohol.

Further, the lower alcohol is 1,2-pentanediol or 1,2-pentanediol and a mixture of other lower alcohols.

Further, the mass ratio of other low-carbon alcohols to 1,2-pentanediol is less than 0.3:1; the other lower alcohol is one or more of ethanol, 1,2-propylene glycol and glycerol.

Further, in the step (1), the mass ratio of the phospholipid to the co-emulsifier is 0.2-1.0: 1.

further, in the step (2), the mass ratio of the emulsifier composition to the oil phase is 1.8:1 to 11:1.

further, in the step (2), the oil phase B is resveratrol or a mixture of resveratrol and liquid oil dissolving resveratrol; the liquid oil dissolving the resveratrol comprises one or more of caprylic/capric triglyceride, squalane, pentaerythritol tetraethyl hexyl ester, castor oil and oleic acid; the mass ratio of the resveratrol in the mixture to the liquid oil for dissolving the resveratrol is 0.1-10: 1.

further, in the step (2), the mass ratio of the oil phase to the water phase is 1:1-23; the mass ratio of the water phase to the oil phase is 1-20: 1, when 1 to 5:1, the obtained inclusion body is in a transparent state, and the ratio of the inclusion body to the inclusion body is 5-20: 1, the obtained inclusion is in a transparent-semitransparent state.

Further, the water phase is pH buffer solution, the pH value is 4-6, and the ionic strength is more than 0.01M.

The particle size of an oil phase in the resveratrol inclusion body is 10-50 nm.

The preparation process of the inclusion is simple and can be realized by stirring at normal temperature.

The resveratrol inclusion body provided by the invention is formed by dispersing an oil phase containing resveratrol in a water phase by an emulsifier according to the diameter of 10-50nm to form a thermodynamically stable inclusion body structure. The inclusion has thermodynamic stability, and can inhibit oxidative degradation and discoloration of resveratrol in air. The inclusion is used as a raw material and directly added into daily chemical products, and can still keep stable inclusion of resveratrol and inhibit discoloration and degradation of resveratrol after being diluted by a water phase.

The beneficial technical effects of the invention are as follows:

(1) The invention realizes the emulsification of the oil phase containing resveratrol at normal temperature by using the phospholipid and the lower alcohol emulsifier composition containing pentanediol to form an inclusion body, and introduces the inorganic salt pH buffering agent to manufacture a fixed pH and ion concentration environment, thereby compressing the electric double layer of the surfactant, leading the resveratrol in the inclusion body to be more stable in wrapping and having the effects of inhibiting degradation and discoloration; and the inclusion bodies thus prepared can be more conveniently used in aqueous formulations to obtain more favoured transparent, translucent personal care products.

(2) The phospholipid used in the invention is a natural surfactant, has good interface performance and self-assembly capacity for forming an interface molecular membrane, and greatly reduces the oil/water interface tension in the presence of the phospholipid according to a negative interface tension forming mechanism formed by an inclusion body in a spontaneous mode, so that the possibility is provided for forming the inclusion body. The presence of the co-emulsifier further reduces the interfacial tension, so that the inclusion can be formed by only stirring at normal temperature without special technology. According to a negative interfacial tension forming mechanism formed by the micro-nano emulsion, the interfacial tension is low but not negative when the phospholipid exists, and the micro-nano emulsion cannot be formed. Further, by introducing the lower alcohol co-emulsifier containing the pentanediol, as the pentanediol molecules are smaller, the pentanediol molecules can be aggregated in a phospholipid interface layer, and the adsorption capacity of the surfactant at the interface is further improved, so that the mixed adsorption of the phospholipid and the pentanediol at the interface can further reduce the interfacial tension to be ultra-low (10) ^-3 -10 ^-5 mN/m), under thermodynamic disturbance to generate transient negative interfacial tension, at the moment, the system spontaneously expands the interface, and then a nano-disperse system with the magnitude of 10nm can be formed. The auxiliary emulsifier can also adjust the interface flexibility, and is beneficial to interface expansion. Compared with other low-carbon alcohols, the pentanediol has the function of adjusting the flexibility of the interface, so that the expansion of the interface is easier to perform during negative interface tension, and the formation of an inclusion is facilitated. The existence of the inorganic salt buffer solution can compress the molecular layer of the phospholipid surfactant, so that the molecular arrangement of the surfactant is more compact, the diffusion of oxygen radicals from the water phase to the oil phase through the molecular layer of the surfactant is further reduced, and the effect of inhibiting the discoloration of the resveratrol is achieved. Therefore, the closer the molecular layer formed by phospholipid and surfactant is, the better the coating effect is, and the lower the degradation rate of the active substance is.

Drawings

FIG. 1 shows the structural formula of resveratrol molecule.

FIG. 2 is a photograph of examples 1 to 5 of the present invention.

In the figure, a is example 1; b. example 2 was used; c. example 3 was used; d. example 4 was used; e. example 5 was used.

FIG. 3 is a photograph of comparative examples 1 to 4 of the present invention.

In the figure, a is comparative example 1; b. comparative example 2; c. comparative example 3; d. comparative example 4 was obtained.

FIG. 4 is a photograph of comparative examples 5 to 10 of the present invention.

In the figure, a is a comparative example 5; b. comparative example 6; c. comparative example 7; d. comparative example 8; c. comparative example 9; d. comparative example 10.

FIG. 5 is a photograph of comparative example 11 of the present invention.

FIG. 6 is a photograph showing the inclusion bodies prepared in examples 1 to 5 of the present invention stored in an incubator at 45 ℃ for 14 days.

In the figure, a is example 1; b. example 2 was used; c. example 3 was used; d. example 4 was carried out; e. example 5 was used.

FIG. 7 is a photograph showing the solutions prepared in comparative examples 1 to 4 of the present invention stored in an incubator at 45 ℃ for 14 days.

In the figure, a is comparative example 1; b. comparative example 2; c. comparative example 3; d. comparative example 4.

FIG. 8 is a photograph showing the solutions prepared in comparative examples 5 to 8 of the present invention stored in an incubator at 45 ℃ for 14 days.

In the figure, a is a comparative example 5; b. comparative example 6; c. comparative example 7; d. comparative example 8.

FIG. 9 is a photograph of a solution prepared in comparative example 11 of the present invention stored in an incubator at 45 ℃ for 14 days.

FIG. 10 is a graph showing the distribution of the particle size of inclusions prepared in examples 1 to 5 of the present invention.

Detailed Description

The present invention will be described in detail with reference to the accompanying drawings and examples.

Example 1:

(1) Adding 1g of soybean phospholipid, 2g of 1,2-pentanediol and 0.3g of ethanol into a 25ml beaker, respectively, and stirring and dissolving uniformly to obtain an emulsifier composition;

(2) Adding 0.1g resveratrol into the emulsifier composition under stirring, mixing with a stirrer, stirring for 10min, adding 0.2g pentaerythritol tetra (ethyl hexanoate) after completely dissolving, and stirring for 10min to obtain a mixture of the emulsifier composition and the oil phase;

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; and (3) adding 6.2g of water phase into the mixture prepared in the step (2) under the stirring condition, and continuously stirring for 10min until the solution is clear and transparent, thus obtaining the inclusion provided by the invention.

Example 2:

(1) Adding 1g of soybean phospholipid, 2g of 1,2-pentanediol and 0.3g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Adding 0.1g of stable emulsifier under stirringResveratrolMixing with a stirrer, stirring for 10min, and adding 0.2g pentaerythritol tetra (ethyl) after completely dissolvingHexanoic acid) Continuously stirring for 10min to obtain mixture of emulsifier composition and oil phase;

(3) Preparing 0.1mol/L NaH ₂ PO ₄ Solution, pH =5 adjusted with phosphoric acid as aqueous phase; and (3) adding 6.4g of water phase into the mixture prepared in the step (2) under the stirring condition, and continuously stirring for 10min until the solution is clear and transparent, thus obtaining the inclusion provided by the invention.

Example 3:

(1) Taking a 25ml beaker, respectively adding 0.44g of soybean lecithin, 2g of 1, 2-pentanediol and 0.2g of isopropanol, and uniformly dissolving to obtain an emulsifier composition;

(2) Adding 0.1g resveratrol into stable emulsifier under stirring, mixing with a stirrer, stirring for 10min, adding GTCC (caprylic/capric triglyceride) 0.2g after completely dissolving, stirring for 10min, and obtaining mixture of emulsifier composition and oil phase after the solution is homogeneous and stable;

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; and (3) adding 6.86g of water phase into the mixture prepared in the step (2) under the stirring condition, and continuously stirring for 10min until the solution is clear and transparent, thus obtaining the inclusion provided by the invention.

Example 4:

(1) Respectively adding 2.3g of lecithin, 2g of 1, 2-pentanediol and 0.3g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Adding 0.1g resveratrol into stable emulsifier under stirring, mixing with a stirrer, stirring for 10min, adding 0.2g squalane after completely dissolving, and stirring for 10min; obtaining a mixture of an emulsifier composition and an oil phase

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; and (3) adding 5.1g of water phase into the mixture prepared in the step (2) under the condition of stirring, and continuing stirring for 10min until the solution is clear and transparent, thus obtaining the inclusion provided by the invention.

Example 5: (example 1 wherein the ratio of the aqueous phase to the oil phase was changed)

(1) Adding 1g of soybean phospholipid, 1.8g of 1, 2-pentanediol, 0.2g of glycerol and 0.3g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Adding 0.1g resveratrol into stable emulsifier under stirring, mixing with a stirrer, stirring for 10min, and adding 0.2g pentaerythritol tetra (ethyl) after completely dissolvingHexanoic acid) Ester stirring for 10min until the solution is uniform and stable; obtaining a mixture of an emulsifier composition and an oil phase

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; and (3) adding 3.6g of water phase into the mixture prepared in the step (2) under the condition of stirring, and continuously stirring for 10min until the solution is clear and transparent, thus obtaining the antioxidant inclusion provided by the invention.

Example 6

(1) Respectively adding 1g of cephalin, 1.8g of 1, 2-pentanediol, 0.2g of glycerol and 0.3g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Adding 2.1g resveratrol into stable emulsifier under stirring, mixing with stirrer, stirring for 10min, and adding 1.2g pentaerythritol tetra (ethyl ester) after completely dissolvingHexanoic acid) Continuing stirring for 10min until the solution is uniform and stable; obtaining a mixture of an emulsifier composition and an oil phase

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 4) as an aqueous phase; and (3) adding 0.3g of water phase into the mixture prepared in the step (2) under the stirring condition, and continuously stirring for 10min until the solution is clear and transparent to obtain the antioxidant inclusion provided by the invention.

Comparative example 1: (converting 1,2 pentanediol to propylene glycol)

A resveratrol inclusion body, the preparation method comprises the following steps:

(1) Adding 1g of soybean phospholipid, 2g of 1,2-propylene glycol and 0.3g of ethanol into a 25ml beaker respectively, and uniformly dissolving to obtain an emulsifier composition;

(2) Under the condition of stirring, adding 0.1g of resveratrol into the stable emulsifier, uniformly mixing by using a stirrer, continuously stirring for 10min, after completely dissolving, adding 0.2g of pentaerythritol tetra (ethyl hexanoate) ester, and continuously stirring for 10min until the solution is uniform and stable;

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; 6.2g of aqueous phase was added to the mixture prepared in step (2) with stirring, and stirring was continued for 10min. Then as shown in fig. 3, inclusions cannot be formed and only an unstable emulsion can be formed.

Comparative example 2: (Co-emulsifier changed to butanediol)

(1) Respectively adding 1g of soybean phospholipid, 2g of 1,2-butanediol and 0.3g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Under the condition of stirring, adding 0.1g of resveratrol into the stable emulsifier, uniformly mixing by using a stirrer, continuously stirring for 10min, after the resveratrol is completely dissolved, adding 0.2g of pentaerythritol tetra (ethyl hexanoate) into the stable emulsifier, and continuously stirring for 10min until the solution is uniform and stable; obtaining a mixture of the emulsifier composition and the oil phase

Comparative example 3: (phospholipid changed to hydrogenated lecithin)

(1) Respectively adding 1g of hydrogenated lecithin, 2g of 1, 2-pentanediol and 0.3g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Adding 0.1g resveratrol into the stable emulsifier under stirring, uniformly mixing by using a stirrer, continuously stirring for 10min, adding 0.2g pentaerythritol tetra (ethyl hexanoate) after the resveratrol is completely dissolved, continuously stirring for 10min, and continuously stirring for 10min to obtain a mixture of an emulsifier composition and an oil phase after the solution is uniform and stable;

Comparative example 4: (use tween instead of phospholipid)

(1) Respectively adding 1g of tween-20, 2g of 1, 2-pentanediol and 0.3g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Under the condition of stirring, adding 0.1g of resveratrol into the stable emulsifier, uniformly mixing by using a stirrer, continuously stirring for 10min, after completely dissolving, adding 0.2g of pentaerythritol tetra (ethyl hexanoate) ester, and continuously stirring for 10min until the solution is uniform and stable; obtaining a mixture of an emulsifier composition and an oil phase

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; 6.2g of an aqueous phase were added to the mixture prepared in step (2) with stirring, and the reaction was continuedStirring for 10min. Then as shown in fig. 3, inclusions cannot be formed and only an unstable emulsion can be formed.

Comparative example 5: (use of pure water instead of pH buffer solution)

(1) Respectively adding 1g of soybean phospholipid, 2g of 1, 2-pentanediol and 0.3g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Adding 0.1g resveratrol into stable emulsifier under stirring, mixing with a stirrer, stirring for 10min, adding 0.2g pentaerythritol tetra (ethyl hexanoate) after completely dissolving, stirring for 10min, adding after the solution is uniform and stable, and stirring for 10min; obtaining a mixture of the emulsifier composition and the oil phase

(3) Taking ultrapure water as a water phase; and (3) adding 6.4g of water phase into the mixture prepared in the step (2) under the condition of stirring, and continuing stirring for 10min until the solution is clear and transparent, thus obtaining the comparative example 5. As can be seen from the degradation tables, comparative example 5 has a higher degradation rate than example, and is heavier in color after storage.

Comparative example 6: (pH of the aqueous phase 7)

(1) Adding 1g of soybean phospholipid, 2g of 1,2-pentanediol, 0.3g of ethanol and 0.2g of AEO9 into a 25ml beaker respectively, and uniformly dissolving to obtain an emulsifier composition;

(2) Under the condition of stirring, adding 0.1g of resveratrol into the stable emulsifier, uniformly mixing by using a stirrer, continuously stirring for 10min, after the resveratrol is completely dissolved, adding 0.2g of pentaerythritol tetra (ethyl hexanoate) into the stable emulsifier, and continuously stirring for 10min until the solution is uniform and stable; obtaining a mixture of an emulsifier composition and an oil phase

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; prepared by the step (2) under stirring6.2g of water phase is added into the mixture, and the mixture is continuously stirred for 10min until the solution is clear and transparent, thus obtaining the comparative example 6. As can be seen from the figure, comparative example 6 was blackened in color after storage.

Comparative example 7: (pH of the aqueous phase 6.5)

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; adding 6.2g of water phase into the mixture prepared in the step (2) under the stirring condition, and continuously stirring for 10min until the solution is clear and transparent to obtain the resveratrol inclusion body. As can be seen from fig. 7, the color change was significant after storage of comparative example 7.

Comparative example 8: (phospholipid to pentanediol ratio 1:9)

A resveratrol solution is prepared by the following steps:

(1) Adding 0.3g soybean phospholipid, 2.7g 1, 2-pentanediol and 0.3g ethanol into 25ml beaker, respectively, and dissolving to obtain emulsifier composition;

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; 6.2g of water phase is added into the mixture prepared in the step (2) under the stirring condition, and the stirring is continued for 10min, so that the system is turbid and can not form inclusion. After storage at 45 ℃, the system is unstable and an oil phase is separated out.

Comparative example 9: (the oil phase and the emulsifier ratio are increased)

A resveratrol solution is prepared by the following steps:

(2) Under the condition of stirring, adding 1g of resveratrol into the stable emulsifier, uniformly mixing by using a stirrer, continuously stirring for 10min, after the resveratrol is completely dissolved, adding 3g of pentaerythritol tetra (ethyl hexanoate) ester, continuously stirring for 10min, adding the solution after the solution is uniform and stable, and continuously stirring for 10min; obtaining a mixture of the emulsifier composition and the oil phase

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; and (3) adding 2.7g of water phase into the mixture prepared in the step (2) under the stirring condition, and continuing stirring for 10min, wherein an oil phase is separated out in the system and an inclusion cannot be formed.

Comparative example 10: (the co-emulsifier is pentanediol and glycerol which are in a mass ratio of 3:1)

A resveratrol solution is prepared by the following steps:

(1) Adding 1g of soybean phospholipid, 1.5g of 1, 2-pentanediol, 0.5g of glycerol and 0.3g of ethanol into a 25ml beaker respectively, and uniformly dissolving to obtain an emulsifier composition;

(2) Under the condition of stirring, adding 0.1g of resveratrol into the stable emulsifier, uniformly mixing by using a stirrer, continuously stirring for 10min, after completely dissolving, adding 0.2g of pentaerythritol tetra (ethyl hexanoate) into the stable emulsifier, and continuously stirring for 10min; obtaining a mixture of an emulsifier composition and an oil phase

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 (pH = 6), and 6.2g of an aqueous phase was added to the mixture prepared in step (2) with stirring as the aqueous phase, and stirring was continued for 10 minutes, whereby the system was turbid and no inclusion was formed.

Comparative example 11: (without addition of phospholipids, a solution system is formed, not an inclusion)

A method for preparing resveratrol hydroalcoholic solution comprises the following steps:

(1) Respectively adding 5g of 1, 2-pentanediol and 2g of ethanol into a 25ml beaker, and uniformly dissolving to obtain an emulsifier composition;

(2) Under the condition of stirring, adding 0.1g of resveratrol into the solvent, and uniformly mixing the resveratrol and the solvent by using a stirrer to obtain a solution; obtaining a mixture of an emulsifier composition and an oil phase

(3) Respectively preparing 0.1mol/L of Na ₂ HPO ₄ With NaH ₂ PO ₄ Solution of Na according to the mass ratio of the solution ₂ HPO ₄ ：NaH ₂ PO ₄ =12:88 to prepare a buffer solution (pH = 6) as an aqueous phase; adding 2.9g of water phase into the mixture prepared in the step (2) under stirring, and continuously stirring for 10min until the solution is clear and transparent to obtain the water-alcohol solution of the uncoated resveratrol.

And (3) performance testing:

(1) And (3) testing color stability:

the inclusion bodies prepared in examples 1-5 and the solutions prepared in comparative examples 1-11 were placed at 45 deg.C, respectivelyConstant temperature boxIn the middle storage, at regular intervalsObserving colourAnd (4) changing. The color change is shown in fig. 2-9. As can be seen, the color stability of the examples is better, while the comparative examples 1-4,8-10 are not able to form stable inclusions. Comparative examples 6 to 7 showed significant discoloration. The pH value of the water phase is proved to have an important effect on the color stability of the resveratrol inclusion body.

(2) And (3) determining the degradation rate of resveratrol:

and (3) measuring the resveratrol degradation rate by using a high performance liquid chromatography-ultraviolet detector. Specifically, the inclusion bodies prepared in examples 1 to 5 and the solution samples prepared in comparative examples 5 to 11 were placed in a 45 ℃ incubator, and at 0d and 28d, samples of a certain concentration were dissolved with methanol, and the content of resveratrol in the samples was measured by high performance liquid chromatography. The HPLC detector used was Waters corporation, USA, and the mobile phase conditions were acetonitrile: water =30, and the selected column was an Agilent C18 column (4.6 mm × 250mm,5 μm) for separation, the column temperature was 30 ℃, the detection wavelength was 306nm, and the degradation rate D of 28D was measured ₂₈ Determined by the following formula:

in the formula: c ₀ Is the resveratrol content in the sample at time 0d, C ₂₈ Is the resveratrol content in the sample at 28 d.

TABLE 1 degradation rates of 28d resveratrol for each sample

	Degradation rate of 28 days	Remarks for note
			Example 1	24.6％	Is composed of
Example 2	30.8％	Is free of
			Example 3	22.9％	Is free of
Example 4	30.5％	Is free of
			Example 5	13.9％	Is free of
Comparative example 5	39.5％	High degradation rate
			Comparative example 6	35.2％	Obvious color change
Comparative example 7	30.4％	Obvious color change
			Comparative example 11	41.3％	High degradation rate

* Note: none of comparative examples 1-4,8-10 formed stable inclusions.

As can be seen from Table 1, the resveratrol system dispersed in the form of solution (comparative example 11) has high degradation rate, and the degradation rate of each example is low, which shows that the phospholipid-based inclusion body has an important effect on the stability of the resveratrol, and the inclusion body technology provided by the invention can effectively inhibit the degradation of the resveratrol. In addition, the degradation rate of the comparative example 5 is also obviously high, which shows that the water phase formed by the pH buffer solution is important for reducing the degradation rate of the resveratrol, and the reason is that a certain ionic strength can compress an electric double layer formed by phospholipid molecules at an oil-water interface, so that the oil-water interface is more compact, oxygen free radicals in water are difficult to transfer into the oil phase, and the oxidative degradation of the resveratrol in the oil phase is inhibited.

FIG. 10 is a distribution diagram of the particle size of the inclusion prepared in examples 1-5 of the present invention, and it can be seen that the resveratrol inclusion provided by the present invention has a small size (10-50 nm), is beneficial to percutaneous absorption and penetration, and has strong application potential.

The above embodiments are merely illustrative of the technical ideas and features of the present invention, and the purpose thereof is to enable those skilled in the art to understand the contents of the present invention and implement the present invention, and not to limit the protection scope of the present invention. All equivalent changes and modifications made according to the spirit of the present invention should be covered within the protection scope of the present invention.

Claims

1. A preparation method of a stabilized resveratrol inclusion body is characterized by comprising the following steps:

2. The method according to claim 1, wherein in the step (1), the phospholipid is one or more selected from soybean phospholipid, lecithin, cephalin and cardiolipin.

3. The method according to claim 1, wherein in the step (1), the co-emulsifier is composed of a lower alcohol.

4. The method of claim 3, wherein the lower alcohol is 1,2 pentanediol or 1,2 pentanediol, and a mixture of other lower alcohols.

5. The method of claim 4, wherein the mass ratio of the other lower alcohols to 1,2-pentanediol is less than 0.3:1; the other low-carbon alcohol is one or more of ethanol, 1,2-propylene glycol and glycerol.

6. The method according to claim 1, wherein in the step (1), the mass ratio of the phospholipid to the co-emulsifier is 0.2 to 1.0:1.

7. the method according to claim 1, wherein in the step (2), the mass ratio of the emulsifier composition to the oil phase is 1.8:1 to 11:1.

8. the preparation method according to claim 1, wherein in step (2), the oil phase B is resveratrol or a mixture of resveratrol and a liquid oil in which resveratrol is dissolved; the liquid oil for dissolving the resveratrol comprises one or more of caprylic/capric triglyceride, squalane, pentaerythritol tetraethyl hexyl ester, castor oil and oleic acid; the mass ratio of the resveratrol to the liquid oil for dissolving the resveratrol in the mixture is 0.1-10: 1.

9. the method according to claim 1, wherein in the step (2), the mass ratio of the oil phase to the water phase is 1:1-23; the water phase is pH buffer solution, and the pH value is 4-6.

10. A stabilized resveratrol inclusion body prepared by the preparation method of any one of claims 1-9, wherein the oil phase in the resveratrol inclusion body has a particle size of 10-50 nm.