CN110787070A

CN110787070A - Preparation method of hemp flower and leaf extract, preparation method of hemp flower and leaf extract and microcapsule, microcapsule and cosmetic

Info

Publication number: CN110787070A
Application number: CN201911010908.XA
Authority: CN
Inventors: 刘秀英
Original assignee: Yunnan Bai Ye Collection Biological Technology Co Ltd
Current assignee: Yunnan Bai Ye Collection Biological Technology Co Ltd
Priority date: 2019-10-23
Filing date: 2019-10-23
Publication date: 2020-02-14

Abstract

The invention relates to the technical field of natural substances, in particular to a preparation method of a hemp flower and leaf extract, a microcapsule preparation method, microcapsules and cosmetics.

Description

Preparation method of hemp flower and leaf extract, preparation method of hemp flower and leaf extract and microcapsule, microcapsule and cosmetic

Technical Field

The invention relates to the technical field of natural substances, in particular to a preparation method of a hemp flower and leaf extract, a preparation method of a hemp flower and leaf extract and microcapsules, microcapsules and cosmetics.

Background

Industrial cannabis refers to a cannabis plant with a tetrahydrocannabinol content of less than 0.3% (dry matter weight%) and its extract product. Industrial Cannabis is known as hemp (hemp) and is a1 year old herbaceous plant of the Cannabis genus (cannabibis) of the Cannabinaceae family (Cannabinaceae), and the now widely used Cannabis is industrial Cannabis.

In the related art, it is difficult to maintain the beneficial components of hemp flower and leaf extract.

Disclosure of Invention

The first object of the present invention is to provide a method for preparing a hemp flower and leaf extract, which can improve the retention of terpenes and cannabidiol in the extract.

The second object of the present invention is to provide a cannabis flower and leaf extract which contains a large amount of terpenes and cannabidiol.

A third object of the present invention is to provide a method for preparing microcapsules capable of protecting terpenes and cannabidiol active ingredients.

A fourth object of the present invention is to provide a microcapsule containing a large amount of terpenes and cannabidiol as active ingredients.

A fifth object of the present invention is to provide a cosmetic composition containing terpenes and cannabidiol, which has a good skin-repairing effect.

The invention is realized by the following steps:

in a first aspect, an embodiment of the present invention provides a method for preparing a cannabis sativa flower and leaf extract, comprising:

mixing pulverized and undried hemp flower and leaf with alcohol solution, and ultrasonic extracting at 60 deg.C or lower to obtain original extract.

In an alternative embodiment, the method further comprises concentrating the raw extract, adding caprylic/capric triglyceride, mixing and dissolving, and separating oil from water to obtain an upper oily extract.

In an alternative embodiment, after the concentrated crude extract is dissolved in caprylic/capric triglyceride, brine is added for dilution, and then standing is carried out until oil-water separation is carried out to obtain an oily extract at the upper layer.

In a second aspect, embodiments of the present invention provide a cannabis flower and leaf extract prepared by the method of preparing a cannabis flower and leaf extract of any one of the preceding embodiments.

In a third aspect, the present invention provides a method for preparing microcapsules, comprising mixing β -cyclodextrin, gum arabic, monoglyceride and the hemp flower and leaf extract of the previous embodiment to prepare an aqueous solution, homogenizing, and spray-drying.

In an alternative embodiment, β -cyclodextrin, gum arabic, monoglyceride and cannabis mosaic extract are mixed to make an aqueous solution, specifically comprising:

mixing β -cyclodextrin solution and acacia solution to obtain glue solution, and mixing the glue solution with the extract of hemp flower and leaf containing monoglyceride.

In a fourth aspect, embodiments of the present invention provide a microcapsule prepared by the method of preparing a microcapsule of the previous embodiments.

In a fifth aspect, embodiments of the present invention provide a cosmetic product comprising a cannabis flower leaf extract of the previous embodiments or microcapsules of the previous embodiments.

In an alternative embodiment, microcapsules are included in an amount of 10-50% of the total cosmetic amount.

In an alternative embodiment, the method comprises the following steps: microcapsules, coenzyme Q10, soy lecithin, tocopheryl acetate, rosemary oil and jojoba oil; alternatively, the first and second electrodes may be,

microcapsules, a thickening agent, a humectant, a solubilizer, essence and a preservative; alternatively, the first and second electrodes may be,

microcapsules, an emulsifier, an emollient, a humectant, an essence and a preservative.

The preparation method of the hemp flower and leaf extract provided by the embodiment of the invention has the beneficial effects that: the preparation method of the hemp flower and leaf extract provided by the invention adopts the undried hemp flower and leaf for extraction, namely, the fresh hemp flower and leaf can be adopted for extraction, and the extraction is carried out by using an alcohol solution at the temperature of less than or equal to 60 ℃, so that the terpenes and the cannabidiol in the hemp flower and leaf are not easy to volatilize and decompose, and the retention amount of the terpenes and the cannabidiol in the extract can be increased.

The hemp flower and leaf extract provided by the embodiment of the invention has the beneficial effects that: the hemp flower and leaf extract provided by the embodiment of the invention is prepared by the method, and the hemp flower and leaf extract has high contents of terpenes and cannabidiol.

The preparation method of the microcapsule provided by the embodiment of the invention has the beneficial effects that the hemp flower and leaf extract is prepared into the microcapsule, the wall material prepared from β -cyclodextrin and Arabic gum can be wrapped by the hemp flower and leaf extract, the loss of terpene and cannabidiol is further reduced, and the active ingredients of terpene and cannabidiol can be protected.

The microcapsule provided by the embodiment of the invention has the beneficial effects that: the microcapsule provided by the embodiment of the invention is prepared by the preparation method of the microcapsule, and contains more terpene and cannabidiol active ingredients.

The cosmetics provided by the embodiment of the invention have the beneficial effects that: the cosmetic provided by the embodiment of the invention comprises the cannabidiol extract or the microcapsule, and further contains terpenes and cannabidiol active substances, so that a good skin repairing effect is achieved.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.

FIG. 1 is a graph showing the results of examination of the effects of macrophage and neutrophil accumulation at the tail fin wound of zebrafish;

FIG. 2 is a graph showing the results of measurements of the effect of macrophages and neutrophils on the clearance of the tail fin wound of zebrafish.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.

The preparation method of the hemp flower and leaf extract, the microcapsule and the cosmetic of the present invention are further described in detail below.

The invention provides a preparation method of a hemp flower and leaf extract, which comprises the following steps: mixing pulverized and undried hemp flower and leaf with alcohol solution, and ultrasonic extracting at 60 deg.C or lower to obtain original extract.

The undried hemp flower and leaf may be fresh hemp flower and leaf. The inventor researches and discovers that when the substances in the hemp flowers and leaves are extracted, if the dried hemp flowers and leaves are used for extraction, the content of terpenes in the extract can be greatly reduced, and more active substances can be effectively reserved by using the hemp flowers and leaves which are not dried, such as: terpenes and cannabidiol.

The inventor researches and discovers that the loss of terpenes and cannabidiol can be further reduced by extracting fresh hemp leaves with an alcoholic solution at the temperature of less than or equal to 60 ℃, so that the prepared extract contains more terpenes and cannabidiol, and the property of the prepared extract is improved.

The research of the inventor finds that the extract with high content of terpene and cannabidiol prepared by the preparation method of the cannabis flower and leaf extract, wherein the terpene can be synergistic with the cannabidiol in effect, so that the effects of the extract such as anti-inflammation and pain relieving are improved.

The alcohol solution may be, for example: 70-95% ethanol solution.

Further, the ratio of the hemp flower and leaf to the alcohol solution may be about 1:5, and the above ratio may be a mass ratio.

The time for ultrasonic extraction of the raw extract of the present invention may be 0.5 to 2 hours, and the intensity of ultrasonic is not particularly limited herein.

The pulverized undried hemp flowers and leaves can be sieved with 20-40 mesh sieve, and then extracted with alcohol solution.

The preparation method of the cannabis sativa extract of the present invention further comprises concentrating the above raw extract and recovering the alcohol solution.

Concentrating the original extract, adding caprylic/capric triglyceride, mixing and dissolving, and separating oil from water to obtain upper oily extract. The upper oily extract is cannabis flos extract containing terpenes and cannabidiol.

Further, after the concentrated crude extract is dissolved in caprylic/capric triglyceride, saline water can be added for dilution, after stirring and mixing, standing, and oil-water separation, the upper layer oily extract, namely the cannabis flower extract containing terpenes and cannabidiol, is obtained, and the oily extract is light green.

The inventor researches and finds that the cannabidiol contained in the oily hemp flower and leaf extract accounts for 5-20% so that the hemp flower and leaf extract meets the requirement of being used as an anti-inflammatory cosmetic additive.

The caprylic acid/capric acid triglyceride is added into the concentrated original extract, so that the fusion performance between the terpene and the cannabidiol can be improved, the synergistic effect of the terpene on the cannabidiol can be further improved, and the performance of the prepared cannabidiol extract is better.

The addition of saline to the mixture of the original extract and caprylic/capric triglyceride for dilution allows the removal of impurities from the extract and allows the rapid separation of the oily extract.

Further, the weight of caprylic/capric triglyceride added to the concentrated crude extract may be 2-5 times that of the concentrated crude extract, so as to make full use of caprylic/capric triglyceride to improve the synergistic effect of terpenes on cannabidiol.

After the concentrated crude extract is mixed with caprylic/capric triglyceride, ultrasonic solubilization can be employed to further utilize caprylic/capric triglyceride to improve the synergistic effect of terpenes on cannabidiol.

The mass concentration of the brine can be about 10%, and the weight or volume of the brine can be 1.5-3 times of the concentrated original extract, so as to rapidly remove impurities and promote oil-water separation.

The preparation method of the microcapsule provided by the invention comprises the steps of mixing β -cyclodextrin, Arabic gum, monoglyceride and the hemp flower and leaf extract prepared by the preparation method of the hemp flower and leaf extract to prepare an aqueous solution, homogenizing, and spray-drying.

Wherein β -cyclodextrin and acacia are used as wall materials of the microcapsule, and monoglyceride is used as an emulsifier of the microcapsule.

Further, the preparation method of the microcapsule specifically comprises the steps of mixing β -cyclodextrin solution and Arabic gum solution to prepare glue solution, and mixing the glue solution with the hemp flower and leaf extract containing monoglyceride.

Further, β -cyclodextrin is dissolved in water to prepare β -cyclodextrin solution, gum arabic is dissolved in water with the temperature of 55-60 ℃ to prepare gum arabic solution, and β -cyclodextrin solution and gum arabic solution are mixed to prepare gum solution.

The monoglyceride contained in the hemp flower and leaf extract containing monoglyceride had a mass concentration of approximately 5%.

The research of the inventor finds that β -cyclodextrin and Arabic gum are used as wall materials of the microcapsules to wrap the hemp flower and leaf extract, so that on one hand, the hemp flower and leaf extract can be protected in the wall materials, volatilization and loss of terpenes and cannabidiol are avoided, more active ingredients are reserved, on the other hand, the solubility of the whole microcapsules can be improved, namely, the solubility of the hemp flower and leaf extract in water is changed, and the subsequent utilization of the hemp flower and leaf extract is facilitated.

The homogenizing method comprises mixing β -cyclodextrin, acacia gum, monoglyceride and the hemp flower and leaf extract prepared by the preparation method of the hemp flower and leaf extract to prepare an aqueous solution, placing the aqueous solution on an oscillator, oscillating for about 48h, controlling the temperature during oscillation to be about 55-60 ℃ and the pressure during oscillation to be about 20MPa, and homogenizing for about 2 times.

The conditions for the spray drying include: the inlet air temperature is 120-140 ℃, the inlet air temperature is controlled to be 55-60 ℃, and the outlet air temperature is controlled to be 60-70 ℃. The inventor researches and discovers that the loss of active substances in the hemp flower and leaf extract can be avoided and the synergistic effect of terpenes in the hemp flower and leaf extract on cannabidiol can be further promoted by adopting the air inlet temperature and the feeding temperature with higher temperature difference.

It should be noted that after spray drying, the microcapsule product may also be sieved through a 70-80 mesh sieve.

The hemp flower and leaf extract and the microcapsules provided by the preparation method of the invention can be used for preparing cosmetics, and the cosmetics can include, but are not limited to, gel, cream, essential oil and the like.

The cosmetic provided by the present invention may be a cosmetic containing 10 to 50% by weight of microcapsules based on the total weight, for example: a gel or a cream.

Further, the microcapsule-containing healing gel may further include: microcapsules, a thickening agent, a humectant, a solubilizer, essence and a preservative. Still further, besides the microcapsule, the microcapsule specifically comprises the following components in percentage by weight: 1 to 10 percent of thickening agent, 5.0 to 40 percent of humectant, 0.5 to 10 percent of solubilizer, 0.2 to 0.8 percent of essence and 0.3 to 0.5 percent of preservative.

The thickening agent is selected from one or more of sodium polyacrylate, carbomer, hydroxyethyl cellulose, polyacrylamide and polyvinyl alcohol; the humectant is one or more selected from sorbitol, glycerol, polyethylene glycol, propylene glycol and xylitol; the solubilizer is selected from one or two of hydrogenated castor oil and PEG-40 hydrogenated castor oil; the antiseptic is selected from one or more of phenoxyethanol, DMDM hydantoin, methyl hydroxybenzoate, propyl hydroxybenzoate, methylisothiazolinone and p-hydroxyacetophenone.

The balance of the above-mentioned microcapsule-containing repair gel is a base gel for preparing a skin care gel.

The microcapsule-containing paste may further include: microcapsules, an emulsifier, an emollient, a humectant, an essence and a preservative. Still further, except the microcapsule, according to the weight percent, specifically include: 5-10% of emulsifier, 2-5% of emollient, 5.0-40% of humectant, 0.2-0.8% of essence and 0.3-0.5% of preservative.

The emulsifier is one or more selected from cetostearyl alcohol, polydimethylsiloxane, fatty acid monoglyceride, behenyl alcohol polyether-25, pentaerythritol distearate and steareth-21. The emollient is selected from one or more of wheat germ oil, grape seed oil, shea butter and sweet almond oil. The humectant is one or more of sorbitol, glycerol, polyethylene glycol, propylene glycol, and xylitol.

The rest of the above-mentioned ointment containing microcapsules is a base cream for preparing a skin care gel.

Essential oil containing hemp flower and leaf extract comprises: microcapsules, coenzyme Q10, soy lecithin, tocopheryl acetate, rosemary oil and jojoba oil; further, except the microcapsule, the microcapsule comprises the following components in percentage by weight: 10.0-60% of hemp flower and leaf extract, coenzyme Q10: 0.5-5%, soybean lecithin 0.5-5%, tocopherol acetate: 0.05-5%, rosemary essential oil: 0.1-5% and the balance of jojoba oil.

The preparation method of the hemp flower and leaf extract, the microcapsule and the cosmetic of the present invention are further described in detail with reference to examples below.

Example 1

Mixing the crushed fresh hemp flower leaves sieved by a 20-mesh sieve with an ethanol solution with the mass concentration of 70%, wherein the mass ratio of the fresh hemp flower leaves to the ethanol solution is 1: 5; then ultrasonic extraction is carried out for 0.5h at the room temperature (28 ℃); concentrating under reduced pressure (temperature not exceeding 60 deg.C), and recovering ethanol to obtain concentrated solution.

Adding caprylic acid/capric acid triglyceride with 2 times of mass into the concentrated solution for dissolving, and using ultrasonic to assist dissolving; adding 10% saline water 1.5 times the mass of the concentrated solution, diluting, stirring, standing for 20min, and oil-water separating to obtain upper oily hemp flower-leaf extract.

β -cyclodextrin is dissolved in water to prepare β -cyclodextrin solution, Arabic gum is dissolved in water with the temperature of 55 ℃ to prepare Arabic gum solution, β -cyclodextrin solution is mixed into the Arabic gum solution, then hemp flower and leaf extract with the mass concentration of 5% is added, water is added and mixed evenly to prepare water solution, wherein the mass ratio of β -cyclodextrin to Ara is 40:3, and the mass ratio of β -cyclodextrin to hemp flower and leaf extract with 5% monoglyceride is 8: 5.

Placing the above aqueous solution on an oscillator, oscillating for 48h at 55 deg.C under 20MPa, and homogenizing for 2 times.

And (3) spray-drying the homogenized mixture, wherein the air inlet temperature is 120 ℃, the feeding temperature is 55 ℃, and the air outlet temperature is 60 ℃. Spray drying, sieving with 70 mesh sieve, and making into microcapsule.

Example 2

Mixing crushed fresh hemp flower leaves sieved by a 40-mesh sieve with an ethanol solution with the mass concentration of 95%, wherein the mass ratio of the fresh hemp flower leaves to the ethanol solution is 1: 5; then ultrasonic extraction is carried out for 2 hours at the temperature of 35 ℃; concentrating under reduced pressure (temperature not exceeding 60 deg.C), and recovering ethanol to obtain concentrated solution.

Adding caprylic acid/capric acid triglyceride with the mass of 5 times into the concentrated solution for dissolving, and using ultrasonic to assist the dissolving; adding saline water with the mass concentration of 10% and the mass amount of 3 times of the concentrated solution, diluting, stirring, standing for 40min, and separating oil from water to obtain upper oily hemp flower and leaf extract.

β -cyclodextrin is dissolved in water to prepare β -cyclodextrin solution, Arabic gum is dissolved in water with the temperature of 60 ℃ to prepare Arabic gum solution, β -cyclodextrin solution is mixed into the Arabic gum solution, then hemp flower and leaf extract with the mass concentration of 5% is added, water is added and mixed evenly to prepare water solution, wherein the mass ratio of β -cyclodextrin to Ara is 40:3, and the mass ratio of β -cyclodextrin to hemp flower and leaf extract with 5% monoglyceride is 8: 5.

Placing the above aqueous solution on an oscillator, oscillating for 52h at 60 deg.C under 20MPa, and homogenizing for 3 times.

And (3) spray-drying the homogenized mixture, wherein the air inlet temperature is 140 ℃, the feeding temperature is 60 ℃, and the air outlet temperature is 70 ℃. After spray drying, sieving with a 80-mesh sieve to obtain the microcapsule.

Example 3

Mixing the crushed fresh hemp flower leaves sieved by a 30-mesh sieve with an ethanol solution with the mass concentration of 80%, wherein the mass ratio of the fresh hemp flower leaves to the ethanol solution is 1: 5.5; then ultrasonic extraction is carried out for 1.5h at the room temperature (25 ℃); concentrating under reduced pressure (temperature not exceeding 60 deg.C), and recovering ethanol to obtain concentrated solution.

Adding caprylic acid/capric acid triglyceride with 3 times of mass into the concentrated solution for dissolving, and using ultrasonic to assist dissolving; adding saline water with mass concentration of 9.5% 2 times of the concentrated solution, diluting, stirring, standing for 30min, and oil-water separating to obtain upper oily hemp flower-leaf extract.

β -cyclodextrin is dissolved in water to prepare β -cyclodextrin solution, Arabic gum is dissolved in water with the temperature of 58 ℃ to prepare Arabic gum solution, β -cyclodextrin solution is mixed into the Arabic gum solution, then hemp flower and leaf extract with the mass concentration of 5% is added, water is added and mixed evenly to prepare water solution, wherein the mass ratio of β -cyclodextrin to Ara is 40:3, and the mass ratio of β -cyclodextrin to hemp flower and leaf extract with 5% monoglyceride is 8: 5.

Placing the above aqueous solution on an oscillator, oscillating for 40h at 58 deg.C under 20MPa, and homogenizing for 2 times.

And (3) spray-drying the homogenized mixture, wherein the air inlet temperature is 130 ℃, the feeding temperature is 58 ℃ and the air outlet temperature is 65 ℃. Spray drying, sieving with 70 mesh sieve, and making into microcapsule.

Example 4

Mixing the crushed fresh hemp flower leaves sieved by a 20-mesh sieve with an ethanol solution with the mass concentration of 90%, wherein the mass ratio of the fresh hemp flower leaves to the ethanol solution is 1: 4.5; then ultrasonic extraction is carried out for 1h at the temperature of 60 ℃; concentrating under reduced pressure (temperature not exceeding 60 deg.C), and recovering ethanol to obtain concentrated solution.

Adding caprylic acid/capric acid triglyceride with the mass of 4 times into the concentrated solution for dissolving, and using ultrasonic to assist the dissolving; adding saline water with mass concentration of 9% 2.5 times of the concentrated solution, diluting, stirring, standing for 40min, and oil-water separating to obtain upper oily hemp flower-leaf extract.

β -cyclodextrin is dissolved in water to prepare β -cyclodextrin solution, Arabic gum is dissolved in 56 ℃ water to prepare Arabic gum solution, β -cyclodextrin solution is mixed into the Arabic gum solution, then hemp flower and leaf extract with the mass concentration of 5% is added, water is added and mixed uniformly to prepare water solution, wherein the mass ratio of β -cyclodextrin to Ara is 40:3, and the mass ratio of β -cyclodextrin to hemp flower and leaf extract with 5% monoglyceride is 8: 5.

Placing the above aqueous solution on an oscillator, oscillating for 50h at 56 deg.C under 20MPa, and homogenizing for 2 times.

And (3) spray-drying the homogenized mixture, wherein the air inlet temperature is 140 ℃, the feeding temperature is 55 ℃, and the air outlet temperature is 65 ℃. After spray drying, sieving with a 80-mesh sieve to obtain the microcapsule.

Examples 5 to 8

Examples 5-8 are repair gels prepared using the microcapsules prepared in examples 1-4, respectively, and methods for preparing the same.

Some of the components of the repair gels provided in examples 5-8 are shown in table 1 below:

TABLE 1

In examples 5 to 8, the respective raw materials were mixed and stirred.

Examples 9 to 12

Examples 9 to 12 are pastes prepared using the microcapsules prepared in examples 1 to 4, respectively, and a method for preparing the same.

Some of the ingredients of the pastes provided in examples 9-12 are shown in table 2 below:

TABLE 2

Examples 9 to 12 were all prepared by mixing and stirring the respective raw materials.

Examples 13 to 16

Examples 13 to 16 are essence oils prepared by using the microcapsules prepared in examples 1 to 4, respectively, and a method for preparing the same.

Examples 13-16 provide essential oils having a portion of the ingredients shown in table 3 below:

TABLE 3

Examples 13 to 16 were all prepared by mixing and stirring the respective raw materials.

First, the repair gel containing 20% microcapsules provided in example 7 was evaluated for anti-inflammatory efficacy, and the test was completed at the university of Guangdong pharmacy.

The zebrafish used in this test were Tg (corola: eGFP) transgenic zebrafish introduced by the university of London, UK, and propagated in this laboratory. Reagent: tricaine (meclin).

The main equipment is as follows: Z-A-D5 five-layer single-row independent culture unit (Shanghai Saint biological laboratory Equipment, Inc.); SZ680 continuously variable magnification microscope (chongqing ott optical instruments ltd); ZXSD-A1090 Biochemical incubator (Shanghai Zhicheng Analyzer manufacturing, Inc.); DMi8 inverted fluorescence microscope (lycra, germany); an electronic scale of one ten thousandth of SQP (sydow scientific instruments (beijing) ltd.).

The main test method comprises the following steps:

1. and feeding the sexually mature zebra fish in the zebra fish culture unit in separate jars. Water temperature: 26 +/-2 ℃; the pH value is 6.7; conductivity: 520 mu s/cm; light/dark cycle: 14h:10 h. The males and females were paired at 1:2 the day before the exposure experiment was started, and naturally mated to lay eggs.

2. Macrophage and neutrophil aggregation assay

1) Healthy zebrafish Tg (corolla: eGFP) developed to 3dpf (days post fertilization) were selected and placed in 6-well cell culture plates, 20 strips/well, and 5mL of test solution was added for pretreatment for 1h, with culture water as a blank control group.

2) Cutting off tail fins of the zebra fish by using a scalpel under a stereomicroscope, putting the zebra fish into a 6-hole cell culture plate, adding 5mL of test solution into each hole of the zebra fish in 15 pieces, and incubating the zebra fish in a curve control biochemical incubator.

3) After incubation for 6h, zebrafish were anesthetized with tricaine, and macrophage and neutrophil accumulation at the tail fin wound was observed under a fluorescent microscope and photographed.

4) The number of cells was counted, and the area within 200 μm from the incision was counted.

3. Macrophage and neutrophil clearance assay

1) Cutting the tail fin of zebra fish Tg (corolla: eGFP) by a scalpel under a stereomicroscope, then placing the zebra fish in a 6-hole cell culture plate, adding 5mL of culture water for culture in 20 strips/hole, and placing the zebra fish in a curve control biochemical incubator for incubation.

2) After incubation for 6h, the zebra fish is placed in a 96-well cell culture plate, and the zebra fish with more macrophage and neutrophil accumulated at the wound is selected under a fluorescence microscope for experiment.

3) The zebra fish is placed in a 6-hole cell culture plate, 15 strips are added in each hole, 5mL of test solution is added, culture water is used as a blank control group, and the zebra fish is placed in a curve control biochemical incubator for continuous incubation.

4) After 6h incubation, namely 12h after tail cutting, the zebra fish is anesthetized by tricaine, and the aggregation of macrophages and neutrophils on the tail fin wound is observed under a fluorescence microscope and photographed and recorded.

5) The number of cells was counted, and the area within 200 μm from the incision was counted.

4. The data were statistically processed using SPSS19.0 software, and the experimental data were expressed as + -SEM data and analyzed by one-way anova. Each concentration group was compared with a blank control group two by two: p < 0.05.

5. 1) see table 4 and the results of fig. 1, the effect of the repair gel (microcapsule content 20%) provided in example 7 on macrophage and neutrophil accumulation at the tail fin wound of zebrafish (compare with blank control: p < 0.05).

TABLE 4 Effect on macrophage and neutrophil accumulation at the tail fin wound of zebrafish (comparison with blank control:. about. P <0.05)

Note: tables 0, 50, 100, 200(μ g/mL) refer to the concentrations after dilution with the repair gel provided in example 7.

As can be seen from fig. 1, the white Control group (Control) showed a large amount of macrophage and neutrophil accumulation at the wound of the tail fin of zebra fish, while the repair gel (microcapsule content: 20%) showed a gradual decrease in macrophage and neutrophil accumulation at the wound of the tail fin of zebra fish.

As can be seen from Table 4 and FIG. 1, the numbers of macrophages and neutrophils in the blank Control group (Control) were 22.25. + -. 1.18. Meanwhile, the number of macrophages and neutrophils in the experimental group with the concentration of 50 mug/mL of the repair gel (the microcapsule content is 20%) is 21.20 +/-0.96, and the number of macrophages and neutrophils in the experimental group is not significantly different from that in the blank control group (22.25 +/-1.18) (P > 0.05). The numbers of macrophages and neutrophils of the experimental groups with the concentration of 100 and 200 mug/mL of the repair gel (the microcapsule content is 20%) are respectively 18.25 +/-0.92 and 18.18 +/-0.70, and the numbers are respectively significant difference (P <0.05) compared with the blank control group (22.25 +/-1.18). Therefore, 100 and 200 mu g/mL of the repair gel (the content of the microcapsule is 20 percent) can obviously inhibit the macrophage and the neutrophil from gathering at the wound of the tail fin of the zebra fish; and it can be determined that the repair gel provided in example 7 can inhibit the accumulation of macrophages and neutrophils at the injury of the tail fin of zebra fish.

2) See table 5 and figure 2 results, the effect of the repair gel provided in example 7 (microcapsule content 20%) on macrophage and neutrophil clearance at the tail fin wound of zebrafish (compare with blank control: p < 0.05).

TABLE 5 Effect of the repair gel (microcapsule content 20%) on macrophage and neutrophil clearance from the injury of the tail fin of zebrafish

(comparison with blank control:. P <0.05)

As can be seen from fig. 2, the white Control (Control) group had a large amount of macrophages and neutrophils aggregated at the wound of the tail fin of zebra fish, while the repair gel (microcapsule content of 20%) group had gradually decreased macrophages and neutrophils aggregated at the wound of the tail fin of zebra fish.

As can be seen from FIG. 2 and Table 5, the number of macrophages and neutrophils in the blank control group was 21.33. + -. 1.11. The numbers of macrophages and neutrophils of the experimental groups with the concentration of 50 and 100 mug/mL of the repair gel (microcapsule content is 20%) are respectively 19.91 +/-1.05 and 18.89 +/-1.10, and no significant difference (P >0.05) is generated compared with the blank control group (21.33 +/-1.11). The number of macrophages and neutrophils in the experimental group with the concentration of the repair gel (the microcapsule content is 20%) being 200 mug/mL is 17.64 +/-0.64, and the number of macrophages and neutrophils in the experimental group is significantly different (P <0.05) compared with that in the blank control group (21.33 +/-1.11). It can be seen that 200 μ g/mL of the repair gel (with a microcapsule content of 20%) significantly promotes the clearance of macrophages and neutrophils at the wound of the tail fin of zebra fish, and it can be determined that the repair gel provided in example 7 can clear the wound of the tail fin of zebra fish.

In conclusion, the repair gel provided by the application has good anti-inflammatory efficacy.

Secondly, the plaster containing 30% of microcapsules provided in example 12 was subjected to a patch test.

The control for this experiment was a cream without microcapsules.

The subjects are 30 persons, 2 persons in male and 28 persons in female, the age is 21-55 years, and the volunteer enrollment criteria of the subjects are met.

The spot test method comprises selecting qualified spot tester, applying 0.03g of the ointment into the spot tester, applying the ointment to the back of the subject with special adhesive tape, and removing the ointment after 24 hr. The skin reactions were observed at 0.5, 24 and 48 hours after the removal, and the results were recorded according to the skin reaction grading standards in technical standards for cosmetic safety (2015 edition), and are shown in table 6.

TABLE 6 test results of human skin patches

Note: the different skin reactions in the above table worsen progressively from 0 to 5.

As is clear from Table 6, the ointment provided by the present invention does not cause adverse reactions on the skin.

Thirdly, the essential oil containing 25% of microcapsules provided in example 15 was subjected to an experiment for preventing and repairing skin barrier damage, the experimental unit is: the Yunnan Huangjia doctor is in the traditional Chinese medicine tumor hospital.

The experiment was carried out according to "national food and drug administration" or "quality control code for non-clinical research on drugs" (9 months in 2003).

The transdermal water loss rate (TEWL) is one of the main methods for judging skin barrier function, and an increase in the transdermal water loss rate (TEWL) can be considered to be associated with a decrease in skin barrier function. It is known that in various problematic skins (e.g., atopic dermatitis, contact dermatitis, etc.) in which the barrier function of the skin is impaired, the loss of moisture from the skin is more vigorous than that of healthy skin.

Therefore, the experiment firstly adopts the test of the percutaneous water loss rate as the judgment of whether the product has the skin barrier damage prevention and repair effect.

The experimental materials and the like mainly comprise: commercial dishwashing detergents: diluting with distilled water to 5% solution according to actual use mass fraction; beaker: 2L specification, production of Beijing glass instruments; a timer: WB388, produced by Shenzhen, Inc. of Eurasia electronics; constant light source cell: the chamber with a constant light source is convenient for a dermatologist to judge skin reaction; a skin moisture content measuring instrument manufactured by CK company of Germany; a water loss rate tester for skin via epidermis, manufactured by CK company of Germany.

1) The method comprises the following steps: the experiment used "repair essential oil (microcapsule content 25%)", a control blank sample (glycerin) for the test. The test is completed by 10 qualified volunteers. Volunteers were between 23 and 41 years of age, with a mean age of 30.2 ± 5.94 years. During the trial, the subjects did not develop erythema, papules, wheal, itching, and other adverse skin reactions associated with the trial samples.

After acclimation in a constant temperature (22 + -1 deg.C) and constant humidity (50 + -5%) room for 30 minutes, the stratum corneum water content of the skin on the inner forearm (divided into two areas) of the volunteer was first tested. The samples were applied (2.0. + -. 0.1) mg/cm2 to an area on the medial aspect of the left forearm (4 cm. times.4 cm) of the volunteer and the stratum corneum hydration of the skin was measured at 1 hour, 2 hours and 4 hours, respectively. The other area was coated with a blank control (glycerol).

The right forearm medial (4cm x 4cm) (divided into two areas) of the volunteers was then tested for transdermal water loss, and the volunteers were peeled off 15 times from the forearm medial side of the right hand with tape to cause skin barrier damage. Samples were applied at (2.0. + -. 0.1) mg/cm in one area of the medial aspect of the right forearm and a blank control (glycerin) applied to the other area and massaged until after absorption, and the skin was tested for transdermal water loss (TEWL) at 1 hour, 2 hours, and 4 hours, respectively.

The evaluation results are shown in Table 7, which shows the average value of the water content of the horny layer.

TABLE 7 stratum corneum Water content at various times (%)

Sample (I)	0 hour	1 hour	2 hours	4 hours
					Essential oil	19.7±8.8	41.5±10.7	40.8±9.5	40.5±9.4
Blank control	19.7±7.3	25.4±7.5	23.3±7.6	22.2±8.8

According to the results in Table 7, the essential oil is larger than the blank control in the effect of increasing the water content of the skin stratum corneum, and the results show that the essential oil has the effect of increasing the water content of the skin stratum corneum.

The evaluation results are shown in Table 8, using the mean value of the percutaneous water loss (TEWL).

TABLE 8 transdermal Water loss rate TEWL/% at different times

Sample (I)	0 hour	1 hour	2 hours	4 hours
					Essential oil	4.4±1.8	6.0±1.7	5.3±0.9	4.5±1.0
Blank control	4.8±1.6	7.3±1.3	6.5±1.0	6.9±1.0

As can be seen from the results in table 8, the essential oil was smaller than the blank control sample in the reduction of the transdermal water loss (TEWL), and the results indicate that the essential oil has the efficacy of preventing and repairing the skin barrier damage.

2) Study subjects: 10 women were enrolled in the trial, aged 30-58 years. Facial performance of all subjects: redness, itching, dryness, molting, etc. The dermatologist selects after clinical evaluation of the facial skin.

Subjects were subjected to clinical evaluation of facial skin by a dermatologist before using the product, and then tested for moisture content and epidermal water loss within 1 hour. The essential oil product is used in the morning and evening, and is continuously used for 28 days, which cannot be interrupted. After 28 days, the clinical evaluation of the facial skin and the test of the moisture content and the water loss rate of the skin through epidermis were carried out in dermatology, and the skin conditions before and after the use were compared. If the skin condition is worsened within 28 days, the patient stops the product, and the serious patient goes to a hospital for a doctor.

The results of the skin moisture test before and after use are compared and shown in Table 9.

Table 9 moisture content (%) of stratum corneum before and after use

The result of measuring the water content of the skin stratum corneum after 28 days of using the essential oil shows that the essential oil has the effect of improving the water content of the skin stratum corneum, and the result shows that the essential oil has the effects of preventing and repairing skin barrier damage.

The results of the determination of the percutaneous water loss (TEWL) in 1 hour of the skin before and after the use are shown in Table 10.

TABLE 10 transdermal Water loss rate TEWL/% of each individual before and after use

After the essential oil is used for 28 days, the reduction effect on the percutaneous water loss rate (TEWL) is obvious, and the result shows that the essential oil has the efficacy of preventing and repairing the skin barrier damage.

In conclusion, the essential oil provided by the invention has the effects of preventing and repairing skin barrier damage.

Example 17

Example 17 microcapsules were prepared similarly to example 3, except that dried hemp flower leaves were used in the extraction of hemp flower leaves.

The microcapsules prepared in example 17 were prepared into essential oil, and the formulation of the essential oil was the same as that of example 15.

The essential oils provided in examples 17 and 15 were subjected to the skin barrier damage prevention and repair test in the same manner as described above, and the results are shown in tables 11 and 12.

TABLE 11 Water content (%) of stratum corneum at various times

Sample (I)	0 hour	1 hour	2 hours	4 hours
					Example 15	19.7±8.8	41.5±10.7	40.8±9.5	40.5±9.4
Example 17	19.7±5.1	37.4±3.3	33.3±1.6	30.2±3.8

TABLE 12 transdermal Water loss rate TEWL/% at different times

Sample (I)	0 hour	1 hour	2 hours	4 hours
					Example 15	4.4±1.8	6.0±1.7	5.3±0.9	4.5±1.0
Example 17	4.8±1.6	6.6±1.1	6.2±0.4	5.4±0.7

From the results in tables 11 and 12, it can be seen that example 15 provides the essential oil larger than example 17 for the effect of increasing the water content of the stratum corneum of the skin; for the reduction of the trans-dermal water loss (TEWL), the essential oil of example 15 is less than that of example 17; the results show that the effect of the microcapsules prepared with the extract extracted from fresh hemp flowers and leaves is better.

In summary, the preparation method of the hemp flower and leaf extract provided by the invention adopts the undried hemp flower and leaf for extraction, namely, the fresh hemp flower and leaf can be adopted for extraction, and the extraction is carried out by using an alcohol solution at the temperature of less than or equal to 60 ℃, so that the terpenes and the cannabidiol in the hemp flower and leaf are not easy to volatilize and decompose, and the retention amount of the terpenes and the cannabidiol in the extract can be increased.

The hemp flower and leaf extract provided by the embodiment of the invention is prepared by the method, and the hemp flower and leaf extract has high contents of terpenes and cannabidiol.

According to the preparation method of the microcapsule provided by the embodiment of the invention, the hemp flower and leaf extract is prepared into the microcapsule, and the hemp flower and leaf extract can be wrapped in β -wall materials prepared from cyclodextrin and Arabic gum, so that the loss of terpenes and cannabidiol is reduced, and the active ingredients of terpenes and cannabidiol can be protected.

The microcapsule provided by the embodiment of the invention is prepared by the preparation method of the microcapsule, and contains more terpene and cannabidiol active ingredients.

The cosmetic provided by the embodiment of the invention comprises the cannabidiol extract or the microcapsule, and further contains terpenes and cannabidiol active substances, so that a good skin repairing effect is achieved.

The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for preparing a hemp flower and leaf extract, comprising:

2. The method of claim 1, further comprising concentrating the crude extract, adding caprylic/capric triglyceride, mixing and dissolving, and separating the mixture into an upper oily extract.

3. The method of claim 2, wherein the concentrated extract is dissolved in the caprylic/capric triglyceride, diluted with saline, and then left to stand for oil-water separation to obtain an oily upper extract.

4. A marijuana flower-leaf extract, which is prepared by the method for preparing a marijuana flower-leaf extract according to any one of claims 1 to 3.

5. A method for preparing microcapsules, which comprises mixing β -cyclodextrin, gum arabic, monoglyceride and the hemp flower and leaf extract of claim 4 to prepare an aqueous solution, homogenizing, and spray-drying.

6. The method for preparing microcapsules of claim 5, wherein β -cyclodextrin, gum arabic, monoglyceride and the hemp flower and leaf extract are mixed to make an aqueous solution, comprising:

mixing β -cyclodextrin solution and acacia solution to obtain glue solution, and mixing the glue solution with the hemp flower and leaf extract containing monoglyceride.

7. A microcapsule produced by the production process for a microcapsule according to claim 5 or 6.

8. A cosmetic comprising the hemp flower and leaf extract of claim 4 or the microcapsule of claim 7.

9. The cosmetic according to claim 8, comprising said microcapsules in an amount of 10 to 50% of the total amount of said cosmetic.

10. The cosmetic according to claim 8, comprising: the microcapsules, coenzyme Q10, soy lecithin, tocopheryl acetate, rosemary oil and jojoba oil; alternatively, the first and second electrodes may be,

the microcapsule, the thickening agent, the humectant, the solubilizer, the essence and the preservative; alternatively, the first and second electrodes may be,

the microcapsule, the emulsifier, the emollient, the humectant, the essence and the preservative.