CN115227649A

CN115227649A - Preparation method of dandelion micro-liposome and dandelion micro-liposome

Info

Publication number: CN115227649A
Application number: CN202210876106.2A
Authority: CN
Inventors: 王福同
Original assignee: Qingdao Pinjian Biotechnology Co ltd
Current assignee: Qingdao Pinjian Biotechnology Co ltd
Priority date: 2022-07-25
Filing date: 2022-07-25
Publication date: 2022-10-25
Anticipated expiration: 2042-07-25
Also published as: CN115227649B

Abstract

The invention discloses a preparation method of dandelion micro-lipid and the dandelion micro-lipid, wherein the preparation method of the dandelion micro-lipid comprises the following steps: s1, extracting dandelion at a low temperature to obtain a low-temperature dandelion extract; s2, concentrating the low-temperature dandelion extract obtained in the step S1 to obtain dandelion concentrated solution; and S3, stirring the dandelion concentrated solution and the lecithin raw material by low-temperature ultrasonic waves until the lecithin raw material completely coats the dandelion concentrated solution to form dandelion liposome. The dandelion liposome obtained by the invention can stably and slowly release the effective components of the low-temperature dandelion extract after being taken by a human body, and improves the oxidation resistance and absorption efficiency of cells of the human body.

Description

Preparation method of dandelion micro-liposome and dandelion micro-liposome

Technical Field

The invention belongs to the technical field of micro-fat, and particularly relates to a preparation method of dandelion micro-fat and the dandelion micro-fat.

Background

According to the records of the literature, dandelion, also known as herba violae and veronicastrum herb, is a common plant in temperate regions to the subtropical regions. The dandelion leaf is rich in vitamin A and vitamin C, and its tender leaf can be used as a wild vegetable. The dandelion has wide medicinal application, such as diuresis, antioxidation, anti-inflammation, ovarian insufficiency treatment, sterility alleviation and climacteric symptom relief, has the bactericidal effect on fungi such as staphylococcus aureus, hemolytic streptococcus and the like, also has the effects of dredging the obstruction of milk vessels and promoting lactation, and is widely used for treating various respiratory diseases and gastroenteritis.

However, the absorption of the vitamin C component contained in the dandelion extract decreases with the increase of the dose, mainly because the absorption of vitamin C is carried out by active transport and passive diffusion transport, the vitamin C is absorbed by the "active transport" at low dose, and when the dose is gradually increased until the "active transport" is saturated, the rest is carried out by the "passive diffusion transport". Therefore, the best mode is "small-amount multiple-time" replenishment to improve the absorption, but slow small-amount multiple-time replenishment is not easy to implement. On the other hand, even if the high dose vitamin C is absorbed into the blood, it is rapidly eliminated by the kidneys and lost from the urine. In addition, some components in the dandelion extract are easily damaged by oxidation, are afraid of heat, acid and alkali and are afraid of light, so that the effective active concentration after the dandelion extract reaches the small intestine is reduced due to interference of a plurality of factors before the dandelion extract reaches the small intestine after being taken, and the absorption efficiency of a human body and the beneficial effect of the dandelion extract on the human body are influenced.

Accordingly, further developments and improvements are still needed in the art.

Disclosure of Invention

The invention aims to provide a preparation method of dandelion micro-lipid and dandelion micro-lipid, wherein dandelion is extracted at low temperature to obtain a low-temperature dandelion extract, the low-temperature dandelion extract is concentrated and then uniformly stirred with a lecithin raw material by using a low-temperature ultrasonic technology, so that the lecithin raw material completely coats the dandelion concentrated solution to form the dandelion micro-lipid, and the dandelion micro-lipid can stably and slowly release effective components of the low-temperature dandelion extract after being taken by a human body, thereby improving the oxidation resistance and absorption efficiency of cells of the human body.

In order to achieve the purpose, the invention provides the following technical scheme:

a preparation method of dandelion liposome comprises the following steps:

s1, extracting dandelion at a low temperature to obtain a low-temperature dandelion extract;

s2, concentrating the low-temperature dandelion extract obtained in the step S1 to obtain dandelion concentrated solution;

and S3, stirring the dandelion concentrated solution and the lecithin raw material by low-temperature ultrasonic wave till the lecithin raw material completely covers the dandelion concentrated solution to form dandelion liposome.

The invention is further configured to: in the step S1, the process of extracting the dandelion at low temperature specifically comprises: mixing dry dandelion materials and water in a weight ratio of 1: extracting at low temperature according to the proportion of 8-22.

The invention is further configured to: the process for extracting the dandelion at the low temperature specifically comprises the following steps: mixing dry dandelion materials and water in a weight ratio of 1:8, low-temperature extraction.

The invention is further configured to: in the step S2, the low-temperature dandelion extract is concentrated to 30% of the original low-temperature dandelion extract by weight percentage to obtain dandelion concentrated solution, wherein the concentration of total flavonoids contained in the dandelion concentrated solution is at least 0.3g/L.

The invention is further configured to: in the step S3, the lecithin raw material consists of lecithin powder and dextrin, and the weight ratio of the dandelion concentrated solution to the lecithin powder to the dextrin is 20: (2-1.2): (1.1-0.7).

The invention is further configured to: in step S3, the stirring time is 30-55 min.

The invention is further configured to: the preparation method of the dandelion liposome further comprises the following steps:

and S4, sieving the dandelion liposome by a homogenizer, wherein the mesh opening of the homogenizer is between 1mm and 1.5 mm.

The invention is further configured to: the preparation method of the dandelion micro-lipid also comprises the following steps:

and S5, sequentially sterilizing and cold-storing the dandelion liposome.

The dandelion liposome prepared by the preparation method comprises a double-layer lecithin membrane layer positioned outside and a dandelion concentrated liquid layer positioned inside, wherein the total flavone contained in the dandelion concentrated liquid layer is at least 0.3g/L.

The invention is further configured to: the particle size of the dandelion liposome is between 1mm and 1.5 mm.

The dandelion liposome prepared by the invention has the following advantages because the dandelion concentrated solution is coated by the lecithin film layer:

(1) The water soluble herba Taraxaci concentrated solution is changed into a stable slow release dosage form, and can increase its absorption efficiency.

(2) The stability of the herba Taraxaci concentrated solution is increased, and the herba Taraxaci concentrated solution is maintained in a state without oxidative damage before cell absorption.

(3) Increase of the ability to cross cell membranes: the double-layer lecithin membrane layer has a structure similar to that of a cell membrane, so that the penetrating power of the double-layer lecithin membrane layer penetrating through the cell membrane and entering cells can be improved, and compared with the traditional intravenous injection, the penetrating power of the double-layer lecithin membrane layer is 8-10 times higher.

(4) The side effect is reduced: by adopting the double-layer lecithin film coating technology, the extract in the dandelion concentrated solution can not generate irritation to the stomach and cause stomach discomfort when being taken.

Drawings

FIG. 1 is a schematic structural diagram of dandelion liposomes in an embodiment of the present invention;

FIG. 2 is a graph showing the DPPH radical scavenging ability of dandelion liposomes in accordance with the present invention;

FIG. 3 is a graph showing the analysis of taraxacum microcapsules stained by cells according to one embodiment of the present invention;

FIG. 4 is a graph showing the absorption rate of dandelion liposomes by cells in accordance with the present embodiment;

in the drawings: 1. dandelion is a micro lipid body; 11. a bilayer lecithin film layer; 12. and (4) a dandelion concentrated liquid layer.

Detailed Description

In order to make those skilled in the art better understand the technical solution of the present invention, the following description is given for clear and complete description of the technical solution of the present invention with reference to the embodiments of the present invention, and other similar embodiments obtained by those skilled in the art without creative efforts based on the embodiments of the present application shall fall within the protection scope of the present application.

Interpretation of terms:

micro-lipid body: liposomes are artificial vesicles in the form of hollow spheres of nanometric dimensions, commonly used in the food industry, which can be suspended in water and are generally made of lipids such as natural phospholipids and cholesterol. Because one end of each molecule of the phospholipid is hydrophilic, the other end of each molecule of the phospholipid is hydrophobic, the phosphate end faces outwards, and the lipid end faces inwards, a lipid sphere with a double-layer lipid film layer as the outer part and a hollow lipid sphere as the inner part is formed immediately after the lipid touches the aqueous solution, wherein the surfaces of the double-layer lipid film layer far away from each other are hydrophilic ends, and the surfaces of the double-layer lipid film layer facing each other in the interlayer are hydrophobic ends, so that the solution of the water-soluble substance can be wrapped in the hollow part in the lipid sphere, and the oil-soluble substance is attached to the lipid film layer.

The invention provides a preparation method of dandelion liposome, which comprises the following steps:

s1, extracting the dandelion at a low temperature to obtain a low-temperature dandelion extract.

Specifically, in step S1, the process of extracting the dandelion at low temperature specifically comprises: mixing dry dandelion materials and water in a weight ratio of 1: and (4) extracting at low temperature according to the proportion of 8-22 to obtain low-temperature dandelion extract. The temperature of the low-temperature extraction is preferably 20 ℃.

S2, concentrating the low-temperature dandelion extract obtained in the step S1 to obtain dandelion concentrated solution.

Concentrating the low-temperature dandelion extract to 30% of the original low-temperature dandelion extract by weight percentage to obtain dandelion concentrated solution, wherein the concentration of total flavonoids contained in the dandelion concentrated solution is at least 0.3g/L.

And S3, stirring the dandelion concentrated solution and the lecithin raw material by low-temperature ultrasonic waves until the lecithin raw material completely coats the dandelion concentrated solution to form dandelion liposome.

Specifically, in step S3, the lecithin raw material is composed of lecithin powder and dextrin, and the ratio of the dandelion concentrated solution to the lecithin powder and the dextrin is 20: (2-1.2): (1.1-0.7), uniformly stirring the dandelion concentrated solution and lecithin raw material prepared from lecithin powder and dextrin for a period of time by low-temperature ultrasonic wave, so that the dandelion concentrated solution is completely coated by the lecithin raw material to form a dandelion micro-lipid body with a double-layer lecithin film layer outside and a dandelion concentrated solution layer inside, and the stirring time is 30-55 min.

Preferably, the temperature of the low-temperature ultrasonic agitation is not higher than 10 ℃.

Preferably, the ratio of the dandelion concentrated solution to the lecithin powder and the dextrin is 20:1.6:0.9, and the stirring time is 45min.

And S5, sequentially sterilizing and cold storing the dandelion micro-lipid.

The step of sterilizing dandelion liposomes and cold storing can be continued after the step of sieving in step S4, or after the step of uniformly stirring the dandelion concentrated solution, lecithin powder and dextrin by low-temperature ultrasonic wave to form dandelion liposomes.

The dandelion liposome 1 obtained by the preparation method has the structure shown in figure 1, and comprises a double-layer lecithin membrane layer 11 positioned outside and a dandelion concentrated liquid layer 12 positioned inside and coated by the double-layer lecithin membrane layer 11, wherein the total flavone contained in the dandelion concentrated liquid layer 12 is at least 0.3g/L. Preferably, the particle size of the dandelion liposomes 1 is between 1mm and 1.5 mm.

The preparation method comprises the steps of extracting dandelion at a low temperature to obtain a low-temperature dandelion extract, concentrating the low-temperature dandelion extract, uniformly stirring the low-temperature dandelion extract with lecithin raw materials by using a low-temperature ultrasonic technology, completely coating the lecithin raw materials with dandelion concentrate to form dandelion micro-fat, and after the dandelion micro-fat is taken by a human body, stably and slowly releasing active ingredients of the low-temperature dandelion extract, so that the oxidation resistance and the absorption efficiency of cells of the human body are improved. Effective components in the dandelion extract are not easy to damage by a low-temperature extraction mode, and the dandelion concentrated solution in the obtained dandelion liposome is in a low-temperature state by a low-temperature ultrasonic technology, so that the effective active concentration of the effective components in the dandelion extract is ensured.

Example 1

A preparation method of dandelion liposome comprises the following steps:

1) Mixing the dried dandelion with water in a ratio of 1:8, performing low-temperature extraction to obtain low-temperature dandelion extract, preferably, the extraction temperature is 20 ℃, and the extraction time is 90 minutes;

2) Concentrating the low-temperature herba Taraxaci extractive solution to 30% of the original low-temperature herba Taraxaci extractive solution to obtain herba Taraxaci concentrated solution containing total flavone at least 0.6g/L;

3) Uniformly stirring the dandelion concentrated solution and a lecithin raw material prepared by lecithin powder and dextrin for a time by using low-temperature ultrasonic waves, wherein the ratio of the dandelion concentrated solution to the lecithin powder to the dextrin is 20:1.6:0.9; completely coating the dandelion concentrated solution with the lecithin raw material to form a dandelion liposome with a double-layer lecithin membrane layer on the outer part and a dandelion concentrated solution layer on the inner part. Wherein the low-temperature ultrasonic stirring time is 45 minutes, and the low-temperature stirring temperature is 10 ℃.

4) Sieving the obtained herba Taraxaci liposome with a homogenizer, wherein the mesh aperture of the homogenizer is 1mm.

5) And (3) sequentially performing sterilization treatment and cold storage treatment on the dandelion liposome after the sieving step, wherein the sterilization treatment time is 5 minutes, the temperature is 90 ℃, and the pasteurization mode is selected. The temperature of the cold storage treatment was 4 ℃. The sterilization and cold storage step can be continued after the sieving step, or after the step of uniformly stirring the dandelion concentrated solution, the lecithin powder and the dextrin by low-temperature ultrasonic waves to form dandelion liposome. In this example, a dandelion liposome was obtained, which comprises a double-layered lecithin film layer located outside and a dandelion concentrated liquid layer located inside and coated with the double-layered lecithin film layer, and the concentration of total flavonoids contained in the dandelion concentrated liquid layer was 0.6g/L. The particle size of the dandelion micro-lipid is 1mm.

When the dandelion liposome is reused as a raw material, the components of the dandelion liposome meet the following standards:

TABLE 1 Standard Table of Taraxacum mongolicum micro-lipid as raw material for reuse

Item	National standard	Inspection method
			Acetone insoluble/% > or more	95	GB1886.238
Water content and volatile matter/% < less	1.5	GB1886.238
			N-hexane insoluble substance/% > is less than or equal to	0.3	GB1886.238
The acid value (calculated by KOH) is less than or equal to mg/kg	38	GB1886.238
			Iodine value/(g/100 g)	60～80	GB1886.238
The peroxide value/(meg/kg) is less than or equal to	50	GB1886.238
			Total arsenic (calculated by As) is less than or equal to mg/kg	3.0	GB1886.238
Lead (Pb)/(mg/kg) is less than or equal to	1.0	GB1886.238

Example 2

A taraxacum liposome was prepared in the same manner as in example 1, except that: the ratio of the dandelion dry material to the water is 1:13, the content of total flavonoids in the dandelion concentrated solution obtained by the same steps as the example 1 is 0.4g/L, namely the content of total flavonoids in dandelion micro-lipid is 0.1g/L.

Example 3

A dandelion liposome prepared by the same preparation method as in example 1, which is different from the dandelion liposome in example 1 in that: the ratio of the dandelion dry material to the water is 1:17, the total flavone content in the dandelion concentrated solution obtained by the same steps as the example 1 is 0.3g/L, namely the total flavone content in the dandelion liposome is 0.1g/L.

Example 4

A dandelion liposome prepared by the same preparation method as in example 1, which is different from the dandelion liposome in example 1 in that: the ratio of the dandelion dry material to the water is 1:22; the total flavone content in the dandelion concentrated solution obtained by the same steps as the example 1 is 0.1g/L, namely the total flavone content in the dandelion micro-lipid is 0.1g/L.

Example 5

A dandelion liposome prepared by the same preparation method as in example 1, which is different from the dandelion liposome in example 1 in that: the ratio of the dandelion dry material to the water is 1:6, at an extraction temperature of 20 ℃. It was found that this example had too high a viscosity to extract and did not give a low temperature dandelion extract with a total flavone content of 0.02g/L in the solution.

The relevant preparation parameter data for examples 1 to 5 are tabulated below:

TABLE 2 preparation of data sheets of parameters for examples 1-5

Performance test

(1) Oxidation resistance test

The ethanol solution of DPPH free radical is dark purple with a maximum absorbance at a wavelength of 517nm, and is used to evaluate the ability of the extract to provide hydrogen. When the sample has the antioxidant capacity, DPPH free radicals can be removed. FIG. 2 is a graph showing the DPPH radical scavenging ability of the dandelion liposomes obtained in examples 1 to 5. Table 2 shows the comparison of the different extraction concentrations and the antioxidant capacities of examples 1 to 5.

TABLE 2 comparison of different extraction concentrations and antioxidant capacities of examples 1-5

As can be seen from fig. 2 and table 2, the dandelion extracts obtained in examples 1 to 4 had increased radical scavenging ability with increasing concentration.

In example 1, the antioxidant capacity of the concentrate was 48% using 100% lecithin extraction, and the antioxidant capacities of examples 2 and 3 were 14% and 10%, respectively, thus demonstrating that example 1 achieved the most optimized antioxidant capacity conditions through a series of process parameters.

In example 1, when the concentration of the lecithin extract concentrate was 25%, the removal rate was 23%, i.e., the effect of the lecithin extract concentrate of 100% in examples 2 and 3 was obtained.

In example 4, when the concentration of the lecithin extract concentrate was 100%, the clearance was 5%, which did not meet the use requirement.

Example 5 is a failure example because the viscosity is too high for extraction in the front end processing.

(2) Cellular uptake assay

Adding red DiI fluorescent dye into lecithin solution to prepare DiI-labeled dandelion liposome. The large intestine epithelial cells were seeded in 6cm petri dishes and after 4 hours replaced with medium containing DiI-labeled dandelion liposomes. The cells and dandelion liposomes were cultured for 0.5 to 8 hours and fixed with 0.05% formaldehyde solution for 30 minutes.

Samples were washed 3 times with buffer solution and stained with the nuclear blue stain DAPI for 5 minutes to calibrate cell locations, recorded and photographed using a fluorescence microscope. The more minute lipid bodies of dandelion are absorbed by cells, the stronger red fluorescence is. Examples 1-5 see fig. 3 and 4, and a table comparing various extraction concentrations with cell uptake rates is shown in table 3.

As can be seen from FIGS. 3, 4 and Table 3, the cell absorption capacity of the resulting dandelion liposomes was different depending on the extraction concentration in the examples. Wherein, the control group is the unit of the number of the cell test basal medium.

TABLE 3 comparison of different extraction concentrations with cellular uptake rates

Examples 1-4 show that the longer the time the cells take up more dandelion liposomes.

Since the cell absorption in example 1 was 17 times that in example 2 and 3 after 8 hours, respectively, 3.6 times and 5 times that in example 1, it was confirmed that the dandelion liposomes had the most optimized cell absorption by the series of process parameters in example 1.

In example 4, the cell absorption after 8 hours was less than 2 times, and the use requirement was not satisfied.

The present invention has been described in detail, and it should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention.

Claims

1. A preparation method of dandelion liposome is characterized by comprising the following steps:

2. The method for preparing dandelion liposomes according to claim 1, wherein in step S1, the process of low temperature extraction of dandelion specifically comprises: mixing dry dandelion materials and water in a weight ratio of 1: low-temperature extraction is carried out according to the proportion of 8 to 22.

3. The method for preparing dandelion liposomes according to claim 2, wherein the process of low temperature extraction of dandelion specifically comprises: mixing dry dandelion materials and water in a weight ratio of 1:8, low-temperature extraction.

4. The method for preparing dandelion liposomes according to claim 1, wherein in step S2, the low temperature dandelion extract is concentrated to 30% of the original low temperature dandelion extract by weight percentage to obtain dandelion concentrated solution, wherein the total flavone concentration in the dandelion concentrated solution is at least 0.3g/L.

5. The method for producing dandelion liposomes according to claim 1, wherein in step S3, said lecithin material is composed of lecithin powder and dextrin, and the ratio of the dandelion concentrate to the lecithin powder and dextrin is 20: (2-1.2): (1.1-0.7).

6. The method for preparing dandelion liposomes according to claim 1, wherein in step S3, the stirring time is 30 to 55min.

7. The method for preparing dandelion liposomes according to claim 1, further comprising the steps of:

8. The method for preparing dandelion liposomes according to claim 1, further comprising the steps of:

and S5, sequentially sterilizing and cold-storing the dandelion liposome.

9. A dandelion liposome prepared by the method for preparing a dandelion liposome according to any one of the claims 1 to 8, wherein the dandelion liposome comprises a double-layer lecithin film layer located outside and a dandelion concentrated liquid layer located inside, and the total flavone contained in the dandelion concentrated liquid layer is at least 0.3g/L.

10. The dandelion liposomes according to claim 9, wherein said dandelion liposomes have a particle size of between 1mm and 1.5 mm.