CN115894597A - Extraction process and application of madecassoside with multiple repair effects - Google Patents

Abstract

The invention discloses an extraction process and application of madecassoside with multiple repairing effects, comprising the following steps: S1, crushing the whole Centella asiatica and extracting it by stirring; Adsorb on the resin column, and elute the active ingredient from the resin column with ethanol; S3, decolorize the medicinal liquid by putting the medicinal liquid on the decolorizing resin column, and the decolorizing liquid is concentrated to a thick paste by a scraper concentrator; S4, go to Drying in a microwave oven in a D-level clean area; S5, crushing, sieving, and total mixing to obtain total asiaticoside; S6, dissolving ethanol to separate asiaticoside to obtain a crude product of madecassoside; S7. Dissolving and refining with 95% ethanol, drying, pulverizing and passing through a sieve to obtain madecassoside. According to the present invention, madecassoside with different soothing and repairing effects is obtained, which is high in purity, safe and non-irritating, and is especially suitable for use in nursing products with moisturizing and multiple repairing effects for children.

Description

A kind of extraction process and application of madecassoside with multiple repairing effects

technical field

The invention relates to the technical field of traditional Chinese medicine preparation, in particular to an extraction process and application of madecassoside with multiple repairing functions.

Background technique

Centella asiatica (L.) Urban is a perennial herb of Umbelliferae, containing amino acids, flavonoids, triterpenoids and other components. Madecassoside (MC) is the pentacyclic triterpenoid with the highest content in Centella asiatica, with a molecular formula of C ₄₈ H ₇₈ O ₂₀ , a relative molecular mass of 975.13, and the appearance of white crystals. Madecassoside is a saponin secondary plant metabolite synthesized by Centella asiatica via the isoprene pathway. Naturally derived compounds usually have a wide range of biological activities in the field of biomedicine, and are also widely used in clinical and daily life.

Research on the function and mechanism of madecassoside in moisturizing, anti-oxidation, and promoting wound healing has been relatively mature. At present, it has been widely used in the production of daily necessities such as wet wipes, skin care products, ointments, and medicines, and in the treatment of skin diseases. Highly effective drug for skin diseases such as psoriasis. In the research field of children's daily skin care, there are still relatively few researches on the efficacy of repairing children's fragile and sensitive skin.

Contents of the invention

In view of the deficiencies in the prior art, the purpose of the present invention is to provide an extraction process and application of madecassoside with multiple repairing effects, and to obtain madecassoside with different soothing and repairing effects. High purity, safe and non-irritating, especially suitable for use in children's moisturizing and multiple repairing care products. In order to achieve the above object and other advantages according to the present invention, a process for extracting madecassoside with multiple repairing effects is provided, including:

S1, pulverizing the whole centella asiatica and stirring and extracting;

S2, through pressure filtration, and carry out resin column adsorption on medicinal liquid, elute active ingredient from resin column with ethanol;

S3. Decolorizing the medicinal liquid by putting the medicinal liquid on the decolorizing resin column, and the decolorizing liquid is concentrated to a thick paste through a scraper concentrator;

S4, transfer to the microwave drying oven in the D-level clean area for drying;

S5, by pulverizing, sieving, and blending, total asiaticosides are obtained;

S6, dissolving and separating asiaticoside in ethanol to obtain a crude product of madecassoside;

S7. Dissolving and refining with 95% ethanol, drying, pulverizing and passing through a sieve to obtain madecassoside.

Preferably, in step S1, the whole centella asiatica is crushed into 60-mesh fine powder, put into extraction tanks, 900 kg of fine powder per tank, and 4.5 tons of boiling water is added to stir and extract for 2 hours.

Preferably, the sieves in step S5 and step S7 are both 100 mesh.

Preferably, the ethanol in step S6 is 5 times 95% ethanol.

Preferably, the mass fraction of the extracted madecassoside is 1%-100%, wherein the product with a mass fraction of 95% madecassoside is added at a concentration of 0.2%-0.04%.

Preferably, the mass fraction of the extracted madecassoside is 1%-10%, wherein the product with a mass fraction of 10% madecassoside is added at a concentration of 0.1%-0.6%.

Application of madecassoside in the preparation of antiallergic products.

Application of madecassoside in the preparation of chemical burn repair products.

Compared with the prior art, the present invention has the beneficial effect that: the madecassoside extracted by the preparation process of the present application can be prepared into madecassoside in any different proportions, and different soothing and repairing effects can be obtained. Madecassoside has high purity, is safe and non-irritating, and is especially suitable for use in children's moisturizing and multiple repairing care products.

Description of drawings

Fig. 1 is the flow chart of the zebrafish embryo neutrophil inhibition test of the extraction process and application of madecassoside with multiple repairing effects according to the present invention;

Fig. 2 is an extraction process of madecassoside with multiple repairing effects according to the present invention and a repairing effect diagram of 95% madecassoside;

Fig. 3 is according to the extracting process of madecassoside with multiple repairing effects according to the present invention and the soothing efficacy test chart with the concentration of Centella asiatica extract applied at 0.1%;

Fig. 4 is a histogram of the soothing efficacy of the extraction process of madecassoside with multiple repairing effects and the applied centella asiatica extract concentration of 0.1% according to the present invention;

Fig. 5 is an extraction process of madecassoside with multiple repairing effects according to the present invention and a test chart of the soothing efficacy of the applied centella asiatica extract at a concentration of 0.3%;

Fig. 6 is a histogram of the soothing effect of the extraction process of madecassoside with multiple repairing effects and the applied centella asiatica extract concentration of 0.3% according to the present invention;

Fig. 7 is an extraction process of madecassoside with multiple repairing effects according to the present invention and a test chart of the soothing effect of the applied centella asiatica extract at a concentration of 0.6%;

Fig. 8 is a histogram of the soothing effect of the extraction process of madecassoside with multiple repairing effects and the applied centella asiatica extract concentration of 0.6% according to the present invention;

Fig. 9 is an extraction process of madecassoside with multiple repairing effects according to the present invention and an experimental diagram of the applied madecassoside in allergy repair;

Fig. 10 is an experimental diagram of the extraction process of madecassoside with multiple repairing effects and the application of madecassoside in chemical burn repair according to the present invention;

Fig. 11 is an extraction process of madecassoside with multiple repairing effects according to the present invention and a chromatogram of 95% madecassoside;

Fig. 12 is a chromatogram of the extraction process of madecassoside with multiple repairing effects and the applied 90% madecassoside according to the present invention.

Detailed ways

The following will clearly and completely describe the technical solutions in the embodiments of the present invention with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only some, not all, embodiments of the present invention. Based on the embodiments of the present invention, all other embodiments obtained by persons of ordinary skill in the art without making creative efforts belong to the protection scope of the present invention.

Referring to Figure 1-12, an extraction process of madecassoside with multiple repairing effects, including: S1, crushing the whole Centella asiatica and stirring to extract;

S2, through pressure filtration, and carry out resin column adsorption on medicinal liquid, elute active ingredient from resin column with ethanol;

S3. Decolorizing the medicinal liquid by putting the medicinal liquid on the decolorizing resin column, and the decolorizing liquid is concentrated to a thick paste through a scraper concentrator;

S4, transfer to the microwave drying oven in the D-level clean area for drying;

S5, by pulverizing, sieving, and blending, total asiaticosides are obtained;

S6, dissolving and separating asiaticoside in ethanol to obtain a crude product of madecassoside;

S7. Dissolve and refine with 95% ethanol, dry, crush and pass through a sieve to obtain madecassoside. According to Figure 11-12, it is the chromatogram of madecassoside, in which the following are 95% centella asiaticoside The peak table of glucoside and 90% madecassoside:

<peak table>

Detector A205nm

peak number keep time area high area% Number of Theoretical Plates (USP) Tailing factor 1 8.537 319085 28428 12.458 12668 1.049 2 9.034 25822 2280 1.008 13583 -- 3 11.384 2004706 141525 78.270 14360 0.983 4 20.817 211642 8746 8.263 16620 1.010 total 2561254 180979 100.000 the

<peak table>

Detector A205nm

peak number keep time area high area% Number of Theoretical Plates (USP) Tailing factor 1 8.868 332269 29064 13.776 13115 0.990 2 11.923 1917930 129295 79.517 14399 0.934 3 22.099 161787 6213 6.708 16276 0.954 total 2411986 164572 100.000 the

Preferably, in step S1, the whole centella asiatica is crushed into 60-mesh fine powder, put into extraction tanks, 900 kg of fine powder per tank, and 4.5 tons of boiling water is added to stir and extract for 2 hours.

Preferably, the sieves in step S5 and step S7 are both 100 mesh.

Preferably, the ethanol in step S6 is 5 times 95% ethanol.

Preferably, the mass fraction of the extracted madecassoside is 1%-100%, wherein the product with a mass fraction of 95% madecassoside is added at a concentration of 0.2%-0.04%.

Preferably, the mass fraction of the extracted madecassoside is 1%-10%, wherein the product with a mass fraction of 10% madecassoside is added at a concentration of 0.1%-0.6%.

Application of madecassoside in the preparation of antiallergic products.

Application of madecassoside in the preparation of chemical burn repair products.

Repair Efficacy Experiment

1. Cosmetic repair effect screening--zebrafish embryo neutrophil inhibition test method

The test method design refers to the OECD TG 236 standard. Test theory basis: BMC Biology.2010,8:151; JLeukoc Biol.2013,94(4):633-642; Molecules.2014,19(2):2390-2409,14:721-721. Before testing, the samples are pre-treated as cosmetic raw material samples according to internal method VI_SOP_002.

1. Terms and definitions

1.1 Zebrafish embryo Zebrafish embryo

A zebrafish embryo at the stage of development that relies on the yolk for energy.

1.2 Lack of heartbeat

Fish embryos whose hearts did not beat for one minute.

1.3 10% lethal concentration 10% Lethal concentration, LC10

The concentration of the test substance that causes the death of 10% of the tested fish embryos. Fish embryos without a heartbeat were judged dead.

2. Principle

The neutrophils of zebrafish embryos and human neutrophils are highly similar in morphology, biochemistry and physiological function. Neutrophils are the first white blood cells to appear at the site of injury or bacterial invasion, and they work to clear infection or harmful substances. The model of neutrophil aggregation caused by copper sulfate-induced neuromast cell damage in the lateral line area of zebrafish embryos was used for testing, and the changes in the number of neutrophils in the lateral line area of fish embryos in the test substance treatment group and the model control group were compared, and calculated Neutrophil inhibition rate is used to evaluate the repair effect screening of raw materials, formulas or products.

3. Instruments and equipment

3.1 Zebrafish farming equipment

The equipment needs to be equipped with temperature control devices, water circulation and filtration devices, and the breeding containers are made of glass or common food-grade PC materials. General zebrafish breeding equipment can be used or fish tanks (such as length, width and height 45cm×45cm×30cm) can be equipped according to laboratory requirements.

3.2 Spawning box/cylinder

Made of glass, stainless steel or other inert materials, equipped with a mesh screen (mesh size is 2mm x 2mm, made of stainless steel or multiple materials to protect fish eggs).

3.3 Water quality monitoring equipment

pH meter, dissolved oxygen meter, salinity meter (conductivity meter).

3.4 Analytical balance

Accuracy 0.1mg.

3.5 Microscope

Equipped with a camera system, the maximum magnification should be greater than 80 times the stereo microscope.

3.6 Test container

Test container: glass or polystyrene test container (eg 6-well plate, Petri dish). If the test substance may be adsorbed on the polystyrene container (such as non-polar chemicals), an inert material (such as glass) should be used to reduce the adsorption.

3.7 Thermostat

Accuracy ±1°C.

3.8 Brine Shrimp Incubator

The equipment needs to be equipped with incubator, air pipe, air volume regulating valve, air pump cut-off switch and check valve device.

3.9 Other routine testing instruments and equipment

Pipettes, centrifuge tubes, glass containers (such as beakers, volumetric flasks, etc.), vortex mixers, ultrasonic water baths, centrifuges, cryogenic refrigerators, adjustable pipettes and tips.

4. Main reagents and consumables

4.1 water

Comply with GB/T 6682 regulations.

4.2 Methanol, dimethyl sulfoxide, ethanol and other co-solvents

Analytical pure level.

4.3 Water for Broodstock Breeding

Weigh 4g of sea salt and dissolve it in 10L of water. The salinity is 0.25-0.50‰, the conductivity is 500-800μS/cm, the saturation of dissolved oxygen is 80%, the pH value is 6.5-8.5, and the hardness is 30-300mg/L (with Calcium Carbonate).

4.4 Eggs of brine shrimp (Artemia salina)

The dormant eggs of brine shrimp need to be kept dry and refrigerated away from light. Weigh about 15g of sea salt and dissolve it in 1L of water to prepare the hatching water for brine shrimp eggs. The density should not exceed 4g/L brine shrimp eggs.

4.5 Fish embryo culture medium

Weigh 2940mg of anhydrous calcium chloride, 1233mg of magnesium sulfate heptahydrate, 630mg of sodium bicarbonate, and 55mg of potassium chloride in 10L of water. The pH value is 6.5-8.5. Chemicals were of analytical grade.

4.6 Copper sulfate solution

Prepare 159.6 mg/L copper sulfate pentahydrate (CuSO4·5H2O) stock solution with water. Store at room temperature.

The concentration of the working solution is 0.1596mg/L, diluted with fish embryo culture medium, and freshly prepared before use.

4.7 Indomethacin solution

Prepare a 35.78 mg/L indomethacin stock solution with dimethyl sulfoxide. refrigeration.

The concentration of the working solution is 0.003578mg/L indomethacin, diluted with fish embryo culture medium, and freshly prepared before use.

4.8 Paraformaldehyde

A solution containing 4% paraformaldehyde and 0.8% sodium hydroxide (NAOH) with a pH of 7.2-7.4 was prepared with phosphate buffered saline (PBS). refrigeration.

4.9 50% and 70% alcohol solutions

Prepare 50% and 70% alcohol solutions with water.

4.10 Sudan black staining solution

Sudan black stock solution: prepare 0.4% Sudan black stock solution with 70% alcohol. refrigeration.

Phenol solution: Dissolve 8g phenol in 15mL ethanol solution, dissolve 0.3g Na2HPO4·12H2O in 50mL water, then mix the two solutions to prepare phenol solution. Store at room temperature, valid within half a year.

Sudan black staining solution was prepared by mixing 900 μL 70% ethanol solution, 100 μL Sudan black stock solution and 1 μL phenol solution.

4.11 PBST

Prepare 0.04% Triton X-100 in PBS solution. Store at room temperature.

4.12 Bleach solution

Prepare _a solution containing 3% _H2O2 and 1% KOH with water. Prepare fresh before use.

4.13 Clear fluid 1

A solution containing 20% glycerol and 0.25% KOH was prepared in water. Store at room temperature.

4.14 Clear fluid 2

A solution containing 50% glycerol and 0.25% KOH was prepared in water. Store at room temperature.

5. Experimental method

5.1 Test organisms

Apply wild-type AB strain zebrafish (Daniorerio) spawning test from a reliable source (such as the National Zebrafish Resource Center of China). Broodstock should have good reproductive capacity – the best age for fish is from 6 to 12 months. Use side crossing as far as possible to maintain genetic diversity in subculture, and a new batch of broodstock should be replaced after 5 generations of pure line broodstock. Broodstock should have no visible signs of infection or disease and have not undergone medical treatment within 2 months. Broodstock should be domesticated for more than 14 days after introduction into the laboratory before breeding and spawning tests.

5.2 Breeding Requirements

5.2.1 The temperature of the breeding water should be controlled at 26-28.5°C, and the indoor temperature should be controlled at 20-25°C.

5.2.2 The breeding density should be controlled at 1 to 2 fish per liter of water, and a fixed 12 to 16 hours of light per day should be maintained, and a good filtration system should be maintained.

5.2.3 Feed at least 2 times a day, including at least 1 brine shrimp (Artemia salina) larvae, and the feeding interval should be more than 3 hours to avoid overfeeding affecting water quality and cleanliness.

5.3 Spawn Requirements

5.3.1 If a spawning tank is used to collect fish eggs, it is advisable to put male fish and female fish into the spawning tank at a ratio of 2:1 1-2 hours before turning off the lights the day before spawning. Since zebrafish do not spawn occasionally, it is recommended to prepare multiple spawning tanks for backup.

5.3.2 In order to avoid genetic deviation, fish eggs collected in at least 3 spawning tanks are mixed and selected for later use. If fish eggs are collected in a breeding fish tank, the egg collection box is put into the breeding fish tank where the eggs are to be collected before turning off the lights on the day before spawning or before turning on the lights on the day of spawning.

5.3.3 In order to prevent fish eggs from being eaten by adult fish, the egg collection box is covered with an inert net.

5.3.4 Mating, spawning and fertilization are completed within about 30 minutes after turning on the light, and the egg collection box can be removed from the fish tank at that time.

5.3.5 After fish eggs are taken out of the egg collection box, it is recommended to clean the fish embryos with fish embryo culture solution.

5.3.6 Select healthy zebrafish embryos and culture them at 28°C until 3 days after fertilization at a density of no more than one fish embryo in 200 μL fish embryo culture medium.

6. Preparation of test substance

6.1 Cosmetic Raw Materials

6.1.1 Water-soluble raw materials: fish embryo culture medium can be used to directly dissolve the test substance to prepare a test solution.

6.1.2 Non-water-soluble raw materials: common solvents such as methanol, dimethyl sulfoxide, ethanol, etc. can be used to aid in dissolution. For example, mix the test substance and organic solvent at a ratio of 1:1 (weight g: volume mL), sonicate for 10 minutes, then add fish embryo culture medium to prepare the required concentration, shake for 30 seconds, and then centrifuge at 6500×g for 10 minutes. Take the supernatant for testing.

6.2 Cosmetic products

6.2.1 Water-soluble products: the test substance can be directly dissolved in fish embryo culture medium to make a test solution. The recommended maximum test concentration is 5g/L.

6.2.2 Non-water-soluble products: common solvents such as methanol, dimethyl sulfoxide, ethanol, etc. can be used to aid in dissolution. For example, mix the test substance and organic solvent at a ratio of 1:1 (weight g: volume mL), sonicate for 10 minutes, then add fish embryo culture medium to prepare the required concentration, shake for 30 seconds, and then centrifuge at 6500×g for 10 minutes. Take the supernatant for testing. The recommended maximum test concentration is 5g/L.

6.3 Precautions

The concentration of the test substance test solution needs to ensure that the mortality rate of zebrafish embryos is 10% (embryos without heartbeat characteristics are defined as dead). Multiple concentration groups can be set as required during testing.

7. Test steps

The following test procedure is a 6-well plate test guideline. Refer to Figure 1 for the testing process. If larger test containers (such as petri dishes) are used, the test guidelines need to be adjusted accordingly. Pick healthy zebrafish embryos that are 3 days post fertilization.

7.1 Test grouping

The test needs to set up a model control group (copper sulfate working solution), a positive control group (copper sulfate working solution + indomethacin working solution) and a test substance group (copper sulfate working solution + test substance).

7.2 Model control group settings

Randomly select 20 fish embryos, transfer them to a 3 cm petri dish, and add 5 mL of fish embryo culture medium containing 0.1596 mg/L copper sulfate pentahydrate (see 4.6 for the preparation method).

7.3 Positive control group settings

Randomly select 20 fish embryos, transfer them to a 3cm petri dish, and add 5mL of fish embryo culture solution containing 0.1596mg/L copper sulfate pentahydrate and 0.003578mg/L indomethacin (see 4.6-4.7 for the preparation method).

7.4 Test substance handling

Randomly select 20 fish embryos, transfer them to a 3 cm petri dish, add 5 mL of fish embryo culture solution containing 0.1596 mg/L copper sulfate pentahydrate and the test substance (see 4.6 and 6.1-6.3 for the preparation method). Place in a 28±1°C incubator and incubate for 40 to 45 minutes.

7.5 Fixed staining of fish embryos

After the fish embryos of each test group were fixed in paraformaldehyde for at least 1 hour, the fish embryos were treated with PBST (see 6.11 for the preparation method) 3 times for 5 minutes each time, and then treated with 50% ethanol (see 4.9 for the preparation method) for 3 minutes. After staining the fish embryos with Sudan black staining solution (see 4.10 for the preparation method) at room temperature for 1 hour, soak them with 70% ethanol (see 4.9 for the preparation method) 4 times, 5 min each time, and then treat them with PBST twice, 5 min each time. Treat the fish embryos with bleach solution for 10 minutes, and keep the lid open if processing in a test tube. Then treat with 70% ethanol solution for 5 minutes, PBST for 1 minute, clear solution 1 (see 4.13 for the preparation method) for 15 minutes, clear solution 2 (see 4.14 for the preparation method) for 10 minutes, and then PBST for 3 minutes.

7.6 Microscopic analysis of samples

Lay the fish embryo sideways. Then put it under a stereo microscope to take pictures of the fish embryo tail.

7.7 Data and calculation of results

Count the number of neutrophils in the lateral line region of the caudal three-quarters from the anus (see Appendix B, Figure B) of each fish embryo.

8. Result calculation

Calculate neutrophil suppression rate:

Neutrophil inhibition rate=(C-T)/C×100% ------------(1)

(1) where:

T—the average value of the number of neutrophils in fish embryos in the test substance treatment group;

C—the average number of neutrophils in fish embryos in the model control group;

The number of neutrophils in fish embryos in the test substance treatment group and the number of neutrophils in the model control group were carried out by two-tailed T-test, and the p value was obtained.

9. Test validity verification

9.1 Test Judgment

At least 90% of the fish embryos in each test group survived after exposure, otherwise the results of the corresponding test group were invalid.

9.2 Test requirements

A positive control group must be set for each batch of tests, and the neutrophil inhibition rate of fish embryos in the positive control group for each batch of tests is required to be positive and p<0.05.

10. Results report

10.1 Neutrophil inhibition rate

The inhibitory rate of the test substance on neutrophils at the corresponding test concentration.

10.2 Evaluation

Evaluate the ability of the test substance to inhibit neutrophils, and analyze whether there is a statistical difference between the test substance treatment group and the model control group (p<0.05).

11. Interpretation of the correlation of results

When the neutrophil inhibition rate is positive and has a statistical difference (p<0.05), the test results can be used as supporting evidence that the test substance has a repairing effect, but it is not a substitute for human efficacy testing.

Notes:

1. Zebrafish embryo neutrophil aggregation model: CuSO4 exposure-induced injury was performed on zebrafish embryos to create a neutrophil aggregation model.

2. Zebrafish embryonic neutrophil inhibition rate=[(number of neutrophils in the model control group-number of neutrophils in the sample treatment group)×100/number of neutrophils in the model control group]%.

3. The p value is obtained from the T-Test analysis of the number of neutrophils in the sample treatment group and the number of neutrophils in the model control group, and it is generally determined that p<0.05 has a statistically significant difference.

4. According to the statistical difference of the inhibition rate and the inhibition rate of the neutrophils in the zebrafish embryo, the repair of the sample

The effect screening was evaluated and divided into significant (inhibition rate was positive and p<0.05) and not significant (inhibition rate was negative or p≥0.05).

The representative pictures of repair effect screening results are as follows:

95% madecassoside was selected as the tested sample, and the test results are shown in Figure 2. In Figure 2, the repair effect is not significant, the repair effect is significant-30% neutrophil inhibition rate, VI-2021455S- 1 Madecassoside 95%: 0.2% formula added concentration, 51% inflammatory cell inhibition rate and VI-2021455S-1 madecassoside 95%: 0.04% formula added concentration, 37% inflammatory cell inhibition rate.

(Whether the following content can be written as an embodiment)

Madecassoside 10% with different concentrations (0.1%, 0.3%, 0.6%) was selected for the same test method, and the following soothing and repairing efficacy data were obtained:

Example 1

Dilute madecassoside 10% to a concentration of 0.1% for soothing and repairing test, the data are as follows:

Remarks: The customer specifies to add 50mL of pure water to each sample bottle to prepare the test sample.

The results of the soothing effect test are shown in Figure 3-4. In Figure 3, the soothing effect is not significant, the soothing effect is significant-30% neutrophil aggregation inhibition rate, VI-20221477S-1 Centella asiatica extract plus10.1%-soothing Significant effect - 46% inhibition rate of neutrophil aggregation, wherein Fig. 4 is a histogram of soothing efficacy test results.

Example 2

Dilute madecassoside 10% to a concentration of 0.3% for soothing and repairing test, the data are as follows:

Remarks: The customer specifies to add 50mL of pure water to each sample bottle to prepare the test sample.

The results of the soothing effect test are shown in Figures 5-6, and Figure 5 is the model control group: the soothing effect is not significant-0% neutrophil aggregation inhibition rate, and the positive control example: 0.0036mg/L indomethacin-significant soothing effect -30% neutrophil aggregation inhibition rate and test sample group: VI-20221477S-2 Centella asiatica extract plusl0.3% - significant soothing effect 39% neutrophil aggregation inhibition rate, columnar in Figure 6 picture.

Example 3

Dilute madecassoside 10% to a concentration of 0.6% for soothing and repairing test, the data are as follows:

Remarks: The customer specifies to add 50mL of pure water to each sample bottle to prepare the test sample.

The results of the soothing effect test are shown in Figure 7-8. Figure 7 is the model control group: the soothing effect is not significant-0% neutrophil aggregation inhibition rate, and the positive control example: 0.0036mg/L indomethacin-significant soothing effect -30% neutrophil aggregation inhibition rate and test sample group: VI-20221477S-3 Centella asiatica extract plusl0.6% - significant soothing effect 41% neutrophil aggregation inhibition rate, columnar in Figure 6 picture.

2. Allergy repair

In this test case, the tester was allergic to dog hair, resulting in irritative hand reactions, redness, swelling, and inflammatory symptoms such as itching. After the tester applied the emulsion formulated with madecassoside 10% for 14 days, the redness and swelling subsided and the itching symptoms also disappeared, as shown in FIG. 9 .

3. Chemical burn repair

In this test case, the tester misused chemical raw materials, resulting in chemical burns, peeling and redness of the cheeks, accompanied by a strong burning sensation. Smear the 5% solution of madecassoside to observe the condition of burn repair.

According to tester feedback:

After 2 days of using a solution of madecassoside, the burning sensation subsided;

After 7 days of use, the redness and swelling of the burn site disappeared;

After 14 days of use, the facial symptoms disappeared and returned to the original state. As shown in Figure 10.

The number of devices and processing scale described here are used to simplify the description of the present invention, and the application, modification and variation of the present invention will be obvious to those skilled in the art.

Although the embodiment of the present invention has been disclosed as above, it is not limited to the use listed in the specification and implementation, it can be applied to various fields suitable for the present invention, and it can be easily understood by those skilled in the art Further modifications can be effected, so the invention is not limited to the specific details and examples shown and described herein without departing from the general concept defined by the claims and their equivalents.

Claims (8)

1. an extraction process of madecassoside with multiple repairing effects, characterized in that, comprising the following steps: S1, pulverizing the whole centella asiatica and stirring and extracting; S2, through pressure filtration, and carry out resin column adsorption on medicinal liquid, elute active ingredient from resin column with ethanol; S3. Decolorizing the medicinal liquid by putting the medicinal liquid on the decolorizing resin column, and the decolorizing liquid is concentrated to a thick paste through a scraper concentrator; S4, transfer to the microwave drying oven in the D-level clean area for drying; S5, by pulverizing, sieving, and blending, total asiaticosides are obtained; S6, dissolving and separating asiaticoside in ethanol to obtain a crude product of madecassoside; S7. Dissolving and refining with 95% ethanol, drying, pulverizing and passing through a sieve to obtain madecassoside. 2. A kind of extraction process of madecassoside with multiple repairing effects as claimed in claim 1, characterized in that, in step S1, the whole plant of Centella asiatica will be crushed to 60 mesh fine powder, and put into the extraction tank In each can of 900kg fine powder, add 4.5 tons of boiling water and stir for extraction for 2 hours. 3. The extraction process of madecassoside with multiple repairing functions as claimed in claim 1, characterized in that the sieves in step S5 and step S7 are both 100 mesh. 4. A process for extracting madecassoside with multiple repairing functions as claimed in claim 1, wherein the ethanol in step S6 is 5 times 95% ethanol. 5. The extraction process of madecassoside with multiple repairing effects as claimed in claim 1, characterized in that the mass of the extracted madecassoside is 1%-100%, wherein the hydroxyl product The mass fraction of madecassoside is 95%, and the added concentration is 0.2%-0.04%. 6. The extraction process of a kind of madecassoside with multiple repairing effects as claimed in claim 5, characterized in that, the quality of the extracted madecassoside is 1%-10%, wherein the hydroxyl product The mass of madecassoside is divided into 10% and the added concentration of the product is 0.1%-0.6%. 7. The use of any madecassoside as claimed in claims 1-6 in the preparation of anti-allergic products. 8. The application of any madecassoside as described in 1-6 of claim 1 in the preparation of anti-chemical burn repair products.

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