CN113456594B - Method for preparing liposome containing glycyrrhiza inflata extract and madecassoside - Google Patents
Method for preparing liposome containing glycyrrhiza inflata extract and madecassoside Download PDFInfo
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Abstract
The invention discloses a preparation method of liposome containing glycyrrhiza inflata extract and madecassoside, which comprises the following steps: A. adding radix Glycyrrhizae Inflatae extract into 1, 3-propylene glycol to obtain solution A; B. adding water and the dissolving solution A into hydroxypropyl-beta-cyclodextrin, and stirring to obtain an aqueous solution of the glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound; C. drying the glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound aqueous solution to obtain an glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound; D. adding lecithin and cholesterol into caprylic/capric triglyceride to obtain oil phase; E. adding the glycyrrhiza inflate extract/hydroxypropyl-beta-cyclodextrin inclusion compound and madecassoside into water to obtain a water phase; F. adding the oil phase into the water phase, and homogenizing and circulating with high pressure homogenizer to obtain liposome containing Glycyrrhrizae radix extract and madecassoside. The invention has the characteristic of effectively improving the bioavailability.
Description
Technical Field
The invention relates to a preparation method of glycyrrhiza inflate root extract/cyclodextrin, in particular to a preparation method of liposome containing glycyrrhiza inflate root extract and madecassoside.
Background
Inflammation is an adaptive reaction generated after the body is damaged or harmfully stimulated, and is closely related to the occurrence and development of a plurality of diseases. Skin inflammation is one of common inflammation problems, the skin is an important protective layer for protecting a human body from external injury and stimulation, when the human body is injured by external stimulation, the immune system of the skin can start signals, inflammatory factors such as arachidonic acid, interleukin and the like and histamine are promoted to be excessively expressed, blood flow is accelerated, permeability of cells is increased, and the skin is caused to have the symptoms of tightness, redness, pruritus, desquamation, even burning, stabbing pain and the like.
The liquorice has a long medicinal history in China and is widely applied to the traditional medicine. The variety of licorice is many, and there are 3 kinds of its original plants recorded in the Chinese pharmacopoeia, namely Ural licorice, Chuangguo licorice and Guangguo licorice. The chemical components of licorice are the main material basis for its medicinal efficacy. Flavonoid compounds, triterpenoid compounds, coumarins, alkaloids, organic acids and other components are separated from liquorice at home and abroad. Licochalcone A is a chalcone compound in Glycyrrhrizae radix, and mainly exists in radix Glycyrrhizae Inflatae. Licochalcone A has multiple biological activities of oxidation resistance, inflammation resistance, bacteria resistance, tumor resistance, immunity regulation and the like, and is widely applied to the industries of food, medicine and cosmetics. However, the glycyrrhiza inflate root extract has poor water solubility, low bioavailability and dark color, so that the application of the glycyrrhiza inflate root extract in cosmetics is limited.
Madecassoside (MA) is a main ingredient of a medicament widely used clinically for treating skin wounds such as surgical wounds, scalds and burns, and has various pharmacological effects of inhibiting bacteria, diminishing inflammation, promoting fibroblast regeneration and the like. However, because it is a polar macromolecular substance, it is not easy to penetrate skin epidermis to reach the lesion site, and the bioavailability is low, so that the function and application of the medicine are limited to a certain extent.
Therefore, the prior art has the problem of low bioavailability.
Disclosure of Invention
The invention aims to provide a preparation method of liposome containing glycyrrhiza inflata root extract and madecassoside. The invention has the characteristic of effectively improving the bioavailability.
The technical scheme of the invention is as follows: a method for preparing liposome containing glycyrrhiza inflata extract and madecassoside comprises the following steps:
A. adding radix Glycyrrhizae Inflatae extract into 1, 3-propylene glycol, and stirring at 30-70 deg.C to obtain solution A;
B. adding water into hydroxypropyl-beta-cyclodextrin, stirring at 30-70 deg.C for dissolving, adding dissolving solution A under stirring, and stirring at 30-70 deg.C for 2-8 hr to obtain radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate water solution;
C. drying the glycyrrhiza inflata extract/hydroxypropyl-beta-cyclodextrin inclusion compound aqueous solution by using a centrifugal spray dryer to obtain an glycyrrhiza inflata extract/hydroxypropyl-beta-cyclodextrin inclusion compound;
D. adding lecithin and cholesterol into caprylic/capric triglyceride, and stirring at 70-90 deg.C to dissolve to obtain oil phase;
E. adding radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate and madecassoside into water, and stirring at 40-80 deg.C to dissolve to obtain water phase;
F. adding the oil phase into the water phase, stirring for 0.5-2h, and homogenizing for 3-10 times by using a high-pressure homogenizer at 500-1500bar pressure to obtain liposome containing the glycyrrhiza inflata extract and the madecassoside.
The preparation method of the liposome containing the glycyrrhiza inflata root extract and the madecassoside comprises the following steps:
A. adding the extract of the root of the glycyrrhiza inflate into 1, 3-propylene glycol, and stirring and dissolving at 50 ℃ to obtain a solution A;
B. adding water into hydroxypropyl-beta cyclodextrin, stirring at 50 deg.C for dissolving, adding dissolving solution A under stirring, and stirring at 50 deg.C for 4 hr to obtain radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate water solution;
C. drying the glycyrrhiza inflata extract/hydroxypropyl-beta-cyclodextrin inclusion compound aqueous solution by using a centrifugal spray dryer to obtain an glycyrrhiza inflata extract/hydroxypropyl-beta-cyclodextrin inclusion compound;
D. adding lecithin and cholesterol into caprylic/capric triglyceride, and stirring at 80 deg.C to dissolve to obtain oil phase;
E. adding radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate and madecassoside into water, and stirring at 80 deg.C to obtain water phase;
F. adding the oil phase into the water phase, stirring for 1 hr, and homogenizing with a high pressure homogenizer at 1000bar for 6 times to obtain liposome containing radix Glycyrrhizae Inflatae extract and madecassoside.
In the preparation method of the liposome containing the glycyrrhiza inflata extract and the madecassoside, when the drying treatment is carried out in the step C, the temperature of an air inlet of a centrifugal spray dryer is 120-170 ℃, and the pump speed of the centrifugal spray dryer is 5-7 r.min -1 。
In the above method for preparing the body containing glycyrrhiza inflata extract and madecassoside paste, the drying treatment in step C is carried out at an air inlet temperature of 160 ℃ and a pump speed of 6.3 r.min -1 。
In the preparation method of the liposome containing the glycyrrhiza inflata root extract and the madecassoside, the step A comprises the following components in parts by weight: 2-8 parts of an extract of glycyrrhiza inflata root; 8-32 parts of 1, 3-propylene glycol.
In the preparation method of the liposome containing the glycyrrhiza inflata root extract and the madecassoside, the step B comprises the following components in parts by weight: 4-16 parts of hydroxypropyl-beta-cyclodextrin; 4-16 parts of water.
In the preparation method of the liposome containing the glycyrrhiza inflata root extract and the madecassoside, the step D comprises the following components in parts by weight: 3-8 parts of lecithin; 1-2 parts of cholesterol; caprylic/capric triglyceride, 15-25 parts.
In the preparation method of the liposome containing the glycyrrhiza inflata root extract and the madecassoside, the step E comprises the following components in parts by weight: 1-5 parts of glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound; 1-5 parts of madecassoside; 55-79 parts of water.
In the preparation method of the liposome containing the glycyrrhiza inflata extract and the madecassoside, the phosphatidylcholine content in the lecithin is more than 75%.
Compared with the prior art, the invention wraps the fat-soluble active substance glycyrrhiza inflate extract in hydroxypropyl-beta-cyclodextrin, the introduction of the hydroxypropyl can break the intramolecular cyclic hydrogen bond of the cyclodextrin, the defect of poor water solubility of the beta-cyclodextrin is overcome while the cavity of the cyclodextrin is kept, thereby the appearance and the solubility of the active substance glycyrrhiza inflate extract can be improved, then the active substance glycyrrhiza inflate extract and the water-soluble active substance madecassoside are simultaneously loaded in the liposome to form the composite cyclodextrin liposome, the transdermal absorption of the composite cyclodextrin liposome is improved, and the bioavailability can be effectively improved. Moreover, the glycyrrhiza inflata root extract and the madecassoside play a role of synergistic increase, so that the glycyrrhiza inflata root extract has better anti-inflammatory effect, and the dosage of each anti-inflammatory substance can be reduced while the anti-inflammatory effect is enhanced, thereby reducing the toxic and side effects caused by high-dosage use.
Meanwhile, the liposome containing the glycyrrhiza inflata extract and the madecassoside, which is prepared by adopting the process steps and parameters of the invention, has uniform particle size, good stability and good transdermal property.
Liposome stability: the particle size distribution and PDI of the liposome containing the glycyrrhiza inflata extract and the madecassoside are measured by a laser particle sizer: the particle size is 120nm, the PDI is 0.23, the particle size is 148nm and the PDI is 0.32 after the liposome containing the glycyrrhiza inflate extract and the madecassoside is placed for 30 days at the temperature of 4 ℃, and the prepared liposome containing the glycyrrhiza inflate extract and the madecassoside has uniform particle size and good stability.
In conclusion, the invention has the characteristic of effectively improving the bioavailability.
Detailed Description
The present invention is further illustrated by the following examples, which are not to be construed as limiting the invention.
Example 1. A method for preparing liposome containing glycyrrhiza inflata root extract and madecassoside comprises the following steps:
A. adding the extract of the root of the glycyrrhiza inflate into 1, 3-propylene glycol, and stirring and dissolving at 30-70 ℃ to obtain a solution A;
B. adding water into hydroxypropyl-beta-cyclodextrin, stirring at 30-70 deg.C for dissolving, adding solution A under stirring, and stirring at 30-70 deg.C for 2-8 hr to obtain radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate aqueous solution;
C. drying the glycyrrhiza inflata extract/hydroxypropyl-beta-cyclodextrin inclusion compound aqueous solution by using a centrifugal spray dryer to obtain an glycyrrhiza inflata extract/hydroxypropyl-beta-cyclodextrin inclusion compound;
D. adding lecithin and cholesterol into caprylic/capric triglyceride, and stirring at 70-90 deg.C to dissolve to obtain oil phase;
E. adding radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate and madecassoside into water, and stirring at 40-80 deg.C to dissolve to obtain water phase;
F. adding the oil phase into the water phase, stirring for 0.5-2h, and homogenizing for 3-10 times by using a high-pressure homogenizer at 500-1500bar pressure to obtain liposome containing the glycyrrhiza inflata extract and the madecassoside.
In the step C, when the drying treatment is carried out, the temperature of the air inlet of the centrifugal spray dryer is 120-170 ℃, and the pump speed of the centrifugal spray dryer is 5-7 r.min -1 。
In the step C, when the drying treatment is carried out, the temperature of an air inlet is 160 ℃, and the pump speed is 6.3 r.min -1 。
The step A comprises the following components in parts by weight: 2-8 parts of glycyrrhiza inflata root extract; 8-32 parts of 1, 3-propylene glycol.
The step B comprises the following components in parts by weight: 4-16 parts of hydroxypropyl-beta-cyclodextrin; 4-16 parts of water.
In the step D, the composition comprises the following components in parts by weight: 3-8 parts of lecithin; 1-2 parts of cholesterol; caprylic/capric triglyceride, 15-25 parts.
The step E comprises the following components in parts by weight: 1-5 parts of glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound; 1-5 parts of madecassoside; 55-79 parts of water.
The phosphatidylcholine content in the lecithin is greater than 75%.
The liposome comprises the following components in parts by weight: 3-8 parts of lecithin; 1-2 parts of cholesterol; caprylic/capric triglyceride, 15-25 parts; 1-5 parts of glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound; 1-5 parts of madecassoside; 55-79 parts of water.
Example 2. A method for preparing liposome containing glycyrrhiza inflata root extract and madecassoside comprises the following steps:
A. adding 5g of radix Glycyrrhizae Inflatae extract into 20g of 1, 3-propylene glycol, and stirring at 50 deg.C to obtain solution A.
B. Adding 10g of water into 10g of hydroxypropyl-beta-cyclodextrin, stirring and dissolving at 50 ℃, adding the dissolving solution A under the stirring condition, and stirring at 50 ℃ for 4 hours to obtain an aqueous solution of glycyrrhiza inflate-beta-cyclodextrin inclusion compound;
C. drying the glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound aqueous solution by using a centrifugal spray dryer, wherein the air inlet temperature is 160 ℃, and the pump speed is 6.3 r.min -1 To obtain the glycyrrhiza inflate extract/hydroxypropyl-beta-cyclodextrin inclusion compound;
D. adding lecithin 5g and cholesterol 1g into caprylic/capric triglyceride 20g, stirring at 80 deg.C to dissolve to obtain oil phase;
E. adding 3g of radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate and 1g of madecassoside into 70g of water, and stirring at 80 deg.C to dissolve to obtain water phase;
F. adding the oil phase into the water phase, stirring for 1 hr, and homogenizing with a high pressure homogenizer at 1000bar for 6 times to obtain liposome containing radix Glycyrrhizae Inflatae extract and madecassoside.
The content of phosphatidylcholine in the lecithin is 80%.
The particle size distribution and PDI of a liposome containing glycyrrhiza inflata root extract and madecassoside prepared in example 2 were measured by a laser particle sizer. Test results show that the particle size of the liposome is 120nm, the PDI is 0.23, the particle size is 148nm and the PDI is 0.32 after the liposome is placed in an environment of 4 ℃ for 30 days, and the prepared liposome containing the glycyrrhiza inflata root extract and the madecassoside has uniform particle size and good stability.
Comparative example 1: 1g of the extract of the root of Glycyrrhiza inflata was dissolved in 4g of 1, 3-propanediol and 1g of madecassoside was dissolved in 94g of water, and the two solutions were mixed uniformly to obtain the product of comparative example 1.
In vitro transdermal experiments: the hair on the dorsal surface of the rat was shaved with a razor and the entire skin was surgically removed. The dermal side was wiped with isopropyl alcohol to remove residual adherent subcutaneous fat. The skin was then repeatedly rinsed with PBS (pH 7.4), wrapped with aluminum foil and stored at-20 ℃ until use. The mouse skin was fixed on Franz diffusion cells with an effective permeation area of 1.766cm 2 The receiving cell was filled with 15mL of PBS buffer, and the temperature of the water bath was 37 ℃. 1mL of the products of example 2 and comparative example 1 were placed in the diffusion cell and sealed. The magnetic stirrer was started and 1.0mL was sampled after 0.5, 1, 2, 4, 6, 8, 12, 16 and 24h, respectively. After sampling, equal volume of constant temperature PBS liquid was supplemented in time. Centrifuging the sample at 10000r/min for 15min, collecting supernatant, measuring the contents of the glycyrrhiza inflate extract and the madecassoside in the sample, and calculating the accumulated permeability.
As a result:
example 2: the cumulative permeability of the madecassoside is 271 mug/cm 2 The cumulative penetration of Glycyrrhiza inflata root extract was 203. mu.g/cm 2 。
Comparative example 1: cumulative permeation of madecassoside: 135 mu g/cm 2 The cumulative penetration of the Glycyrrhiza inflata root extract was 152. mu.g/cm 2 。
Synergistic anti-inflammatory assay
Comparative example 2: liposome of glycyrrhiza inflate root extract
The preparation method comprises the following steps: the remaining steps and components were the same as in example 2, except that: 6g of glycyrrhiza inflate/hydroxypropyl-beta-cyclodextrin inclusion compound was added in step E of example 2 without adding madecassoside.
Comparative example 3: hydroxyasiaticoside liposome
The preparation method comprises the following steps: the remaining steps and components were the same as in example 2, except that: in step E, 2g of madecassoside is added without adding the glycyrrhiza inflate/hydroxypropyl-beta-cyclodextrin inclusion compound.
The specific method of the synergistic anti-inflammatory test is as follows: LPS is adopted to induce a RAW264.7 mouse macrophage inflammation model for evaluation, RAW264.7 cell strain in logarithmic growth phase is taken and inoculated on a 6-well plate at the density of 2 multiplied by 106/mL, and the administration is carried out after the incubation in an incubator for 24 h. The blank group was added with complete medium, the model group was added with LPS at a mass concentration of 5mg/L, the administration group was added with test sample at a mass concentration of 100. mu.g/mL and LPS at a mass concentration of 5mg/L, respectively, to make the total volume of each group 2mL, and the cells were placed in a cell incubator for further incubation for 24 hours.
Cell supernatants were collected and assayed exactly as described in the NO detection kit instructions, with each experiment repeated 3 times independently. Calculating the NO inhibition rate: the inhibition rate is (model group NO content-sample group NO content)/(model group NO content-blank group NO content) × 100%, and the synergy index of each component is calculated from the inhibition rate.
Calculation of synergy index (Berenbaum index) SI:
As a result:
NO inhibition ratio of example 2: 39.29 percent
NO inhibition ratio of comparative example 2: 14.29 percent
NO inhibition ratio of comparative example 3: 25.00 percent
The synergistic index is 0.5057<1, which indicates that the glycyrrhiza inflate extract and the madecassoside have synergistic anti-inflammatory effect.
Claims (9)
1. A preparation method of liposome containing glycyrrhiza inflata root extract and madecassoside is characterized by comprising the following steps:
A. adding the extract of the root of the glycyrrhiza inflate into 1, 3-propylene glycol, and stirring and dissolving at 30-70 ℃ to obtain a solution A;
B. adding water into hydroxypropyl-beta-cyclodextrin, stirring at 30-70 deg.C for dissolving, adding solution A under stirring, and stirring at 30-70 deg.C for 2-8 hr to obtain radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate aqueous solution;
C. drying the glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound aqueous solution by using a centrifugal spray dryer to obtain a glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound;
D. adding lecithin and cholesterol into caprylic/capric triglyceride, and stirring at 70-90 deg.C to dissolve to obtain oil phase;
E. adding radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate and madecassoside into water, and stirring at 40-80 deg.C to dissolve to obtain water phase;
F. adding the oil phase into the water phase, stirring for 0.5-2h, and homogenizing for 3-10 times by using a high-pressure homogenizer at 500-1500bar pressure to obtain liposome containing the glycyrrhiza inflata extract and the madecassoside.
2. The method for preparing liposomes containing glycyrrhiza inflata extract and madecassoside as claimed in claim 1, comprising the steps of:
A. adding the extract of the root of the glycyrrhiza inflate into 1, 3-propylene glycol, and stirring and dissolving at 50 ℃ to obtain a solution A;
B. adding water into hydroxypropyl-beta cyclodextrin, stirring and dissolving at 50 ℃, adding the dissolving solution A under the stirring condition, and stirring at 50 ℃ for 4 hours to obtain an aqueous solution of the glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound;
C. drying the glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound aqueous solution by using a centrifugal spray dryer to obtain a glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound;
D. adding lecithin and cholesterol into caprylic/capric triglyceride, and stirring at 80 deg.C to dissolve to obtain oil phase;
E. adding radix Glycyrrhizae Inflatae extract/hydroxypropyl-beta-cyclodextrin clathrate and madecassoside into water, and stirring at 80 deg.C to dissolve to obtain water phase;
F. adding the oil phase into the water phase, stirring for 1 hr, and homogenizing with a high pressure homogenizer at 1000bar for 6 times to obtain liposome containing radix Glycyrrhizae Inflatae extract and madecassoside.
3. The method for preparing liposome containing glycyrrhiza inflata root extract and madecassoside according to claim 1 or 2, wherein: in the step C, when the drying treatment is carried out, the temperature of the air inlet of the centrifugal spray dryer is 120-170 ℃, and the pump speed of the centrifugal spray dryer is 5-7 r.min -1 。
4. The method for producing a lipid containing glycyrrhiza inflata root extract and madecassoside according to claim 3, wherein: in the step C, when the drying treatment is carried out, the temperature of an air inlet is 160 ℃, and the pump speed is 6.3 r.min -1 。
5. The method for preparing liposome containing glycyrrhiza inflata root extract and madecassoside according to claim 1 or 2, wherein the step a comprises the following components in parts by weight: 2-8 parts of glycyrrhiza inflata root extract; 8-32 parts of 1, 3-propylene glycol.
6. The method for preparing liposome containing glycyrrhiza inflata root extract and madecassoside according to claim 1 or 2, wherein the step B comprises the following components in parts by weight: 4-16 parts of hydroxypropyl-beta-cyclodextrin; 4-16 parts of water.
7. The method for preparing liposome containing glycyrrhiza inflata root extract and madecassoside according to claim 1 or 2, wherein the step D comprises the following components in parts by weight: 3-8 parts of lecithin; 1-2 parts of cholesterol; caprylic/capric triglyceride, 15-25 parts.
8. The method for preparing liposome containing glycyrrhiza inflata root extract and madecassoside according to claim 1 or 2, wherein the step E comprises the following components in parts by weight: 1-5 parts of glycyrrhiza inflata root extract/hydroxypropyl-beta-cyclodextrin inclusion compound; 1-5 parts of madecassoside; 55-79 parts of water.
9. The method of claim 1 or 2, wherein the phosphatidylcholine content in the lecithin is greater than 75%.
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| CN107744502A (en) * | 2017-09-08 | 2018-03-02 | 华南理工大学 | A kind of madecassoside liposome of high encapsulation rate and high stability and preparation method and application |
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