CN110755322A

CN110755322A - Jindan anti-allergy traditional Chinese medicine extract and preparation method and application thereof

Info

Publication number: CN110755322A
Application number: CN201911142287.0A
Authority: CN
Inventors: 张靖华; 张和平; 王莉娜; 王一宇; 刘畅
Original assignee: Shanghai Huayimei Cosmetics Co Ltd
Current assignee: Shanghai Guoyi Kangmei Health Management Co ltd
Priority date: 2019-11-20
Filing date: 2019-11-20
Publication date: 2020-02-07

Abstract

The invention discloses a preparation method of a Jindan anti-allergy traditional Chinese medicine extract, which comprises the following steps: (1) the raw material medicaments in parts by weight are crushed: 15-30 parts of scutellaria baicalensis, 6-15 parts of radix sophorae flavescentis, 5-10 parts of coptis chinensis, 3-8 parts of phellodendron amurense, 10-20 parts of rhizoma smilacis glabrae, 15-30 parts of honeysuckle, 8-20 parts of peony root bark, 5-15 parts of fructus cnidii and 5-15 parts of fructus kochiae, then uniformly mixing, and adding a proper amount of solvent for soaking to obtain a soaking solution; (2) carrying out low-temperature ultrasonic extraction on the soak solution, and carrying out centrifugal separation, microfiltration membrane filtration and reverse osmosis membrane pressurization concentration on the obtained ultrasonic extracting solution to obtain a refined extracting solution; (3) drying the refined extract to obtain powder or adding appropriate amount of 1, 3-propylene glycol, 1, 3-butanediol or glycerol. The Jindan anti-allergy traditional Chinese medicine extract disclosed by the invention has a remarkable synergistic effect on anti-inflammatory and anti-allergy effects, can be used for effectively relieving and treating the hormone-dependent dermatitis, has the effects of relieving and resisting oxidation, and is particularly suitable for people with sensitive skin, weak skin barrier and hormone-dependent dermatitis.

Description

Jindan anti-allergy traditional Chinese medicine extract and preparation method and application thereof

Technical Field

The invention belongs to the technical field of cosmetics, and relates to a Jindan anti-allergy traditional Chinese medicine extract and a preparation method and application thereof.

Background

The skin is used as the first major organ of a human body and plays an important role in the health of the human body, and according to the domestic epidemiological survey, the incidence rate of allergic diseases in China in 2010 is 37.3%, wherein the incidence rate of sensitive muscle groups is about 3%, and the sensitive muscle groups are increased at a rate of 23 times per 10 years. Sensitive Skin (SS) is a highly reactive, poorly tolerated or even Sensitive Skin, i.e. a condition of subjective sensory symptoms induced by susceptibility to various factors. It is widely accepted in medicine that SS is not a skin disorder, but is a skin condition. With the increasing environmental pollution, the development of cosmetic diversity and the improvement of people's awareness of skin health, the perception rate and the attention of SS are gradually rising.

Skin irritation is ICD. ICD occurs through the pathogenic pathways of stimulus entering the stratum corneum of the skin to cause the activation or slight damage of keratinocytes, thereby producing inflammatory cytokines and the like, which are expressed as skin erythema, edema and desquamation. Skin allergy, ACD, includes 2 phases of the priming and priming phases. In the sensitization phase, after the skin is firstly contacted with hapten (stimulus), the skin is transplanted into lymph from the epidermis to sensitize T lymphocytes to form effector T cells and memory T cells, and then the effector T cells and the memory T cells are spread through the blood stream and the whole body. During the priming phase, when the skin is again exposed to the same hapten, memory T cells rapidly differentiate into a number of effector T lymphocytes, triggering an inflammatory response and cytotoxicity. Most consumers use skin care products to protect the skin from damage, and from allergies. However, the components of surfactants, preservatives, essences and the like commonly used in cosmetics are all easy to be used as stimulus sources to stimulate the skin and cause skin irritation or allergy. According to the diagnosis standard and treatment principle of cosmetic contact dermatitis, the common allergens of the cosmetic contact dermatitis reach more than 130. Some cosmetic consumers have a thin horny layer on the face, so that the moisture retention capacity is reduced, sebum secretion is less, the skin is dry, allergy is easy to cause after the skin is contacted with the cosmetic, the face skin is fragile, and the face is easily affected by external environments or substances such as cold air, food, water quality, ultraviolet rays, synthetic fibers, fragrance, pigments and the like, and the hormone face caused by red swelling and pruritus appears. At present, the attention of consumers on the stimulation and sensitivity caused by cosmetics per se reaches an unprecedented level, and the safety of the cosmetics is ranked at the top of four characteristics of the cosmetics.

In summary, the addition of anti-allergic components to cosmetics to reduce the irritation and allergic effects of the cosmetics to the skin is an effective method. With the development of traditional Chinese medicine extraction technology, more and more traditional Chinese medicine extracts are applied to cosmetics as antiallergic components. Although many plant extracts are claimed to have anti-allergy effects, it is unclear whether these traditional Chinese medicines can extract effective anti-allergy components in sufficient amounts to enable the effective anti-allergy components to produce good anti-allergy effects when added into cosmetics. Meanwhile, most of the traditional Chinese medicine extracts added into the existing cosmetics are extracts of single traditional Chinese medicine, and the anti-allergy effect is not good enough. How to compound a multi-component Chinese herbal compound and how to extract effective anti-allergy active ingredients in the Chinese herbal compound most effectively by using any extraction method still needs to be deeply researched. The anti-allergy effect is improved on the basis of the existing traditional Chinese medicine extract, and even if the anti-allergy effect is improved by 10%, the actual application value and the economic benefit which can be brought are very obvious.

Therefore, there is a need to develop a compound herbal extract that can be used in cosmetics to produce anti-allergy effects.

Disclosure of Invention

The first objective of the present invention is to provide an anti-allergic traditional Chinese medicine extract containing jindan, so as to overcome the above technical problems in the prior art.

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

the preparation method of the jindan anti-allergy traditional Chinese medicine extract comprises the following steps:

(1) the raw material medicaments in parts by weight are crushed: 15-30 parts of scutellaria baicalensis, 6-15 parts of radix sophorae flavescentis, 5-10 parts of coptis chinensis, 3-8 parts of phellodendron amurense, 10-20 parts of rhizoma smilacis glabrae, 15-30 parts of honeysuckle, 8-20 parts of peony root bark, 5-15 parts of fructus cnidii and 5-15 parts of fructus kochiae, then uniformly mixing, and adding a proper amount of solvent for soaking to obtain a soaking solution;

(2) carrying out low-temperature ultrasonic extraction on the soak solution in the step (1), and carrying out centrifugal separation, microfiltration membrane filtration and reverse osmosis membrane pressurization concentration on the obtained ultrasonic extracting solution to obtain a refined extracting solution;

(3) drying the refined extracting solution in the step (2) into powder to obtain the jindan anti-allergy traditional Chinese medicine extract; or adding appropriate amount of plant-derived 1, 3-propylene glycol, 1, 3-butanediol or glycerol into the refined extractive solution of step (2) to obtain JINDANKANGMIN Chinese medicinal extract.

As is readily understood by those skilled in the art, the plant-derived 1, 3-propanediol, 1, 3-butanediol or glycerol is used as a natural preservative and humectant, and the mass ratio of the 1, 3-propanediol, 1, 3-butanediol or glycerol to the refined extract is usually 1 (1-3). In the present invention, the mass ratio of the 1, 3-propanediol, 1, 3-butanediol or glycerol to the refined extract is preferably 1 (1-2).

Preferably, in the step (1), the raw material medicines in parts by weight include: 20-25 parts of scutellaria baicalensis, 10-14 parts of radix sophorae flavescentis, 5-10 parts of coptis chinensis, 3-5 parts of phellodendron amurense, 10-15 parts of rhizoma smilacis glabrae, 15-18 parts of honeysuckle, 8-13 parts of peony root bark, 6-10 parts of fructus cnidii and 5-8 parts of fructus kochiae.

Further preferably, in the step (1), the raw material medicines in parts by weight include: 20 parts of scutellaria baicalensis, 14 parts of sophora flavescens, 8 parts of coptis chinensis, 4 parts of phellodendron amurense, 12 parts of rhizoma smilacis glabrae, 18 parts of honeysuckle, 13 parts of peony root bark, 6 parts of fructus cnidii and 5 parts of fructus kochiae.

Preferably, in the step (1), the solvent is water or ethanol water solution; the mass ratio of the raw material medicine to the solvent is 1 (10-50), and the raw material medicine is soaked for 1-3 hours.

Preferably, in the low-temperature ultrasonic extraction step, the ultrasonic temperature is: 25-50 ℃, the ultrasonic frequency is 20-50 kHz, and the single ultrasonic time is as follows: 15 min-30 min; the ultrasonic frequency is as follows: 3 times to 5 times.

Under the conditions, the effective components in the raw material medicines are fully dissolved, the effectiveness of the active components is ensured, and the content of the active components in the traditional Chinese medicine extract is improved.

Preferably, the microfiltration membrane filtration step adopts a microfiltration membrane with the diameter of 0.1-1 μm for filtration to obtain a filtrate L; and in the reverse osmosis membrane pressurization concentration step, the filtrate L is concentrated to 20-50% of the volume of the filtrate L through a reverse osmosis membrane under 0.5-3 MPa.

Preferably, the volume percentage of the ethanol in the ethanol water solution is 20-80%.

Under the conditions, the effectiveness of the active components is further ensured, and the content of the active components of the traditional Chinese medicine extract is improved.

The raw materials can adopt various commercially available products. Preferably, the pretreatment method of part of the raw material is as follows:

coptis chinensis: slicing Coptidis rhizoma, adding succus Zingiberis recens, stirring, moistening, placing into a parching container after succus Zingiberis recens is absorbed completely, heating with slow fire, parching to dry, taking out, and cooling. The inhibiting effect on Proteus is enhanced after processing with ginger juice.

Scutellaria baicalensis: grading the size of cleaned Scutellariae radix, heating in water, softening, cutting into pieces, and drying.

Honeysuckle flower: taking honeysuckle flower to dry, wherein the initial drying temperature is 30-35 ℃, the temperature is raised to 40 ℃ after 2h of initial drying, the drying is carried out for 5-10 h, then the room temperature is controlled to be kept at 45-50 ℃, the temperature is raised to 55-60 ℃ after 10h of drying, and the flower is dried rapidly.

Peony root bark: taking peony roots, removing fine roots, peeling roots, drying in the sun, removing impurities, cleaning with water, moistening thoroughly, slicing, drying, and screening to remove scraps.

The second purpose of the invention is to provide the jindan anti-allergy traditional Chinese medicine extract obtained by the preparation method of the jindan anti-allergy traditional Chinese medicine extract.

The third purpose of the invention is to provide the application of the jindan anti-allergy traditional Chinese medicine extract in preparing anti-allergy cosmetics or anti-allergy ointment.

Further preferably, the antiallergic cosmetic or the antiallergic ointment consists of the jindan anti-allergy traditional Chinese medicine extract and auxiliary materials, wherein the content of the jindan anti-allergy traditional Chinese medicine extract is 1-40 wt%, and the content of the auxiliary materials is 60-99 wt%.

Further preferably, the antiallergic cosmetics include face toilet, face cleanser, face mask, cream, essence water, toner, and toner.

Compared with the prior art, the invention has the following beneficial technical effects:

the gold lead anti-allergy traditional Chinese medicine extract disclosed by the invention is prepared by compounding a plurality of traditional Chinese medicines and extracting the gold lead anti-allergy traditional Chinese medicine extract by a specific extraction process, and the obtained gold lead anti-allergy traditional Chinese medicine extract has a remarkable synergistic effect on anti-inflammatory and anti-allergy effects, can be used for effectively relieving and treating hormone-dependent dermatitis, has a soothing and anti-oxidation effect, and is particularly suitable for people with sensitive skin and weak skin barrier and people suffering from hormone-dependent dermatitis.

The 1, 3-propylene glycol, 1, 3-butylene glycol or glycerin of plant sources can play a role of a preservative, and no preservative is required to be additionally added, so that the Jindan anti-allergy traditional Chinese medicine extract disclosed by the invention is less irritant, and is further beneficial to anti-allergy.

Drawings

FIG. 1 shows the cytotoxicity activity test of the Jindan anti-allergy Chinese medicine extract.

FIG. 2 is a graph of the local skin changes on the face of a severely red volunteer at initial use, 2 and 4 weeks.

FIG. 3 is a graph of facial local skin changes in facial hormone facial severe volunteers at initial, 2, 4 weeks of use.

Detailed Description

The present invention will be further described with reference to the following examples. It should be understood that the following examples are illustrative only and are not intended to limit the scope of the present invention.

Pretreatment of part of raw material medicine

(1) And washing the raw materials for 3 times to effectively remove impurities such as silt, dust and the like.

(2) And coptis chinensis: taking clean coptis root, slicing, adding ginger juice, uniformly stirring (12.5 kg of ginger or 4kg of dried ginger is used for juicing or decocting every 100kg of coptis root), moistening, putting the rhizoma zingiberis into a frying container after the ginger juice is absorbed completely, heating the rhizoma zingiberis in a slow fire, frying the rhizoma zingiberis dry, taking out the rhizoma zingiberis, and cooling the rhizoma zingiberis. The inhibiting effect on Proteus is enhanced after processing with ginger juice.

(3) And scutellaria baicalensis: grading the size of clean Scutellariae radix, heating with water, steaming to round gas for 0.5 hr, taking out, cutting into 1-1.5mm pieces, and drying at 80 deg.C. Or decocting Scutellariae radix in boiling water for 5-10 min, taking out, sealing for 8-12 hr, cutting into 1-1.5mm pieces when the humidity is consistent, and drying at 80 deg.C.

(4) And honeysuckle flower: drying flos Lonicerae at initial drying temperature of 30-35 deg.C for 2 hr, heating to 40 deg.C, drying for 5-10 hr, maintaining at room temperature of 45-50 deg.C, drying for 10 hr, heating to 55-60 deg.C, and rapidly drying. The flos Lonicerae extractive solution contains various tannins for regulating cortex secretion, and has effects of keeping moisture and whitening skin.

(5) And the root of peony: taking peony roots, removing fine roots, peeling off root barks, drying in the sun, removing impurities, cleaning with water, moistening thoroughly, slicing, drying, and screening to remove scraps.

Jindan anti-allergy traditional Chinese medicine extract formula

The formula of the Jindan anti-allergy traditional Chinese medicine extract is shown in table 1.

TABLE 1 anti-allergic Chinese medicine extract formula of Jindan

Example 1 preparation of anti-allergic traditional Chinese medicine extract of Jindan

The preparation method of the jindan anti-allergy traditional Chinese medicine extract adopts a formula 1 of the jindan anti-allergy traditional Chinese medicine extract, and comprises the following steps:

(1) crushing the raw material medicines according to the formula amount into particles, sieving the particles with a 200-400-mesh sieve, uniformly stirring and mixing the particles to obtain mixed particles, adding an ethanol water solution (50% by volume of ethanol) with the mass ratio of 1:10, and soaking the mixed particles for 2 hours to obtain a soaking solution;

(2) carrying out ultrasonic extraction on the soaking solution obtained in the step (1), wherein the ultrasonic frequency is 30kHz, and the ultrasonic temperature is as follows: 30 ℃, the ultrasonic time is 30min, and the ultrasonic frequency is as follows: centrifuging for 3 times (centrifuging speed 3000rpm for 20 min), and collecting supernatant to obtain ultrasonic extractive solution;

(3) filtering the ultrasonic extracting solution in the step (2) by using a 0.5-micron microporous filter membrane, concentrating the obtained filtrate L to 50% of the volume of the filtrate L by using a reverse osmosis membrane under the pressure of 1.5MPa, and evaporating at 40 ℃ to remove ethanol to obtain a refined extracting solution;

(4) adding 1, 3-propylene glycol of plant origin into the refined extracting solution obtained in the step (3), wherein the mass ratio of the refined extracting solution to the 1, 3-propylene glycol is 1:2, and the obtained traditional Chinese medicine extracting solution is the Jindan anti-allergy traditional Chinese medicine extract without preservatives and with pure plants.

Or drying the refined extract in the step (3) to obtain powdery jindan anti-allergy traditional Chinese medicine extract, and storing.

Example 2 preparation of anti-allergic traditional Chinese medicine extract of Jindan

The preparation method of the jindan anti-allergy traditional Chinese medicine extract adopts a formula 2 of the jindan anti-allergy traditional Chinese medicine extract, and comprises the following steps:

(1) crushing the raw material medicines according to the formula amount to particles, sieving the particles with a 200-400-mesh sieve, stirring and uniformly mixing the particles to obtain mixed particles, adding deionized water with the mass ratio of 1:20, and soaking the particles for 3 hours to obtain a soaking solution;

(2) carrying out ultrasonic extraction on the soaking solution obtained in the step (1), controlling the ultrasonic frequency to be 50kHz, and controlling the ultrasonic temperature to be: ultrasonic treatment time of 30min at 45 ℃, ultrasonic frequency: centrifuging for 5 times (centrifuging speed 3000rpm for 20 min), and collecting supernatant to obtain ultrasonic extractive solution;

(3) filtering the ultrasonic extracting solution in the step (2) by using a 1-micron microporous filter membrane, and concentrating the obtained filtrate L to 20% of the volume of the filtrate L under the pressure of 2MPa by using a reverse osmosis membrane to obtain a refined extracting solution;

(4) adding 1, 3-butanediol of plant source into the refined extracting solution obtained in the step (3), wherein the mass ratio of the refined extracting solution to the 1, 3-butanediol is 1:1, and the obtained traditional Chinese medicine extracting solution is the Jindan anti-allergy traditional Chinese medicine extract without preservative and with pure plant.

Example 3 preparation of anti-allergic traditional Chinese medicine extract of Jindan

In the embodiment, a compound traditional Chinese medicine extract is prepared by adopting a Jindan anti-allergy traditional Chinese medicine extract formula 3, and the method comprises the following steps:

(1) crushing the raw material medicines according to the formula amount into particles, sieving the particles with a 200-400-mesh sieve, stirring and uniformly mixing the particles to obtain mixed particles, adding an ethanol water solution (70% by volume of ethanol) with the mass ratio of 1:20, and soaking the mixed particles for 1 hour to obtain a soaking solution;

(2) carrying out ultrasonic extraction on the soaking solution obtained in the step (1), wherein the ultrasonic frequency is 40kHz, and the ultrasonic temperature is as follows: 25 ℃, the ultrasonic time is 15min, and the ultrasonic frequency is as follows: centrifuging for 3 times (at 3000rpm for 20 min), and collecting supernatant to obtain ultrasonic extractive solution;

(3) filtering the ultrasonic extracting solution in the step (2) by using a 0.5-micron microporous filter membrane, concentrating the obtained filtrate L to 50% of the volume of the filtrate L by using a reverse osmosis membrane under the pressure of 1.5MPa, and evaporating at 48 ℃ to remove ethanol to obtain a refined extracting solution;

(4) adding 1, 3-propylene glycol of plant origin into the refined extracting solution obtained in the step (3), wherein the mass ratio of the refined extracting solution to the 1, 3-propylene glycol is 1:1.5, and the obtained traditional Chinese medicine extracting solution is the jindan anti-allergy traditional Chinese medicine extract without preservatives and pure plants.

The final mass of the Chinese medicinal extractive solution prepared in the above examples 1-3 was about 700-750 g.

Comparative example 1 preparation of Single-component extract

Extracting Scutellariae radix 100g in formula 1, adding ethanol water solution (50% by volume of ethanol) at a mass ratio of 1:10, and extracting with the same preparation method as example 1 to obtain refined extractive solution, and adding plant-derived 1, 3-propylene glycol to obtain Scutellariae radix extractive solution. Or directly drying the refined extractive solution to obtain Scutellariae radix extract.

Extracting 100g of radix sophorae flavescentis in the formula 1, adding an ethanol water solution (70% by volume of ethanol) in a mass ratio of 1:10 to extract, and adding plant-derived 1, 3-propylene glycol into the obtained refined extract to obtain the radix sophorae flavescentis extract by the same preparation method as the example 1. Or directly drying the refined extractive solution to obtain radix Sophorae Flavescentis extract.

Extracting Coptidis rhizoma 100g in formula 1, adding ethanol water solution (50% by volume of ethanol) at a mass ratio of 1:10, and extracting with the same preparation method as example 1 to obtain refined extractive solution, and adding plant-derived 1, 3-propylene glycol to obtain Coptidis rhizoma extractive solution. Or directly drying the refined extractive solution to obtain Coptidis rhizoma extract.

Extracting cortex Phellodendri 100g in formula 1, adding ethanol water solution (50 vol% ethanol) at a weight ratio of 1:10, and extracting with the same preparation method as in example 1 to obtain refined extractive solution, and adding plant-derived 1, 3-propylene glycol to obtain cortex Phellodendri extractive solution. Or directly drying the refined extractive solution to obtain cortex Phellodendri extract.

Extracting 100g of rhizoma smilacis glabrae in the formula 1, adding an ethanol water solution (50% by volume of ethanol) in a mass ratio of 1:10 to extract, and adding plant-derived 1, 3-propanediol into the obtained refined extracting solution to obtain the rhizoma smilacis glabrae extracting solution by the same preparation method as the example 1. Or directly drying the refined extractive solution to obtain rhizoma Smilacis Glabrae extract.

Extracting 100g of honeysuckle flower in the formula 1, adding an ethanol water solution (50% by volume of ethanol) with the mass ratio of 1:10 to extract, and adding plant-derived 1, 3-propylene glycol into the obtained refined extract to obtain the honeysuckle flower extract in the same preparation method as the example 1. Or directly drying the refined extractive solution to obtain flos Lonicerae extract.

Extracting 100g of peony root bark in the formula 1, adding an ethanol water solution (50% by volume of ethanol) in a mass ratio of 1:10 to extract, and adding plant-derived 1, 3-propanediol into the obtained refined extract to obtain a peony root bark extract in the same preparation method as the example 1. Or directly drying the obtained refined extract to obtain the peony root-bark extract.

The method comprises the steps of extracting 100g of fructus cnidii in the formula 1, adding an ethanol water solution (50% by volume of ethanol) in a mass ratio of 1:10, and extracting, wherein the other preparation methods are the same as those of the example 1, and adding plant-derived 1, 3-propylene glycol into the obtained refined extracting solution to obtain the fructus cnidii extracting solution. Or directly drying the refined extractive solution to obtain fructus Cnidii extract.

Extracting fructus Kochiae 100g in formula 1, adding ethanol water solution (50% by volume of ethanol) at a mass ratio of 1:10, and extracting with the same preparation method as example 1 to obtain refined extractive solution, and adding plant-derived 1, 3-propylene glycol to obtain fructus Kochiae extractive solution. Or directly drying the refined extractive solution to obtain Kochiae fructus extract.

Comparative example 2 preparation of Jindan anti-allergy Chinese medicine extract by conventional method

The jindan anti-allergy traditional Chinese medicine extract is prepared by a conventional method, and the specific method comprises the following steps:

mixing the raw material medicines of the jindan anti-allergy traditional Chinese medicine extract in the formula 1, adding deionized water in a mass ratio of 1:40, directly decocting for 3 hours, filtering, adding an appropriate amount of 50% ethanol water solution in volume percentage into the filtrate for alcohol precipitation, filtering, and evaporating ethanol to obtain a refined extracting solution;

drying the refined extract to obtain powdered Jindan anti-allergy Chinese medicinal extract; or adding a proper amount of 1, 3-propylene glycol into the refined extracting solution, wherein the mass ratio of the refined extracting solution to the 1, 3-propylene glycol is 1:2, so as to obtain the jindan anti-allergy traditional Chinese medicine extract.

Example 4 detection of active substance content in Chinese medicine

Method for detecting content of total flavonoids as active substances of traditional Chinese medicine

The detection of the total flavone is carried out according to the first part P30 of Chinese pharmacopoeia by referring to the method for measuring the total flavone in hawthorn leaves in the pharmacopoeia.

(1) And preparing a total flavone reference substance solution.

Precisely weighing 25mg of reference substance (rutin) dried at 120 deg.C to constant weight, placing in a 50ml measuring flask, adding appropriate amount of ethanol, ultrasonic treating to dissolve, cooling, adding ethanol to scale, and shaking. Precisely measuring 20ml, placing into a 50ml measuring flask, adding water to scale, and shaking to obtain total flavone reference solution (each 1ml contains anhydrous rutin 0.2 mg).

(2) And preparing a standard curve.

Precisely measuring 1mL, 2mL, 3mL, 4mL, 5mL and 6mL of the reference substance solution prepared in the step (1), respectively placing the reference substance solution in a 25mL measuring flask, adding water to 6mL, adding 1mL of 5% sodium nitrite solution, shaking up, placing for 6 minutes, adding 1mL of 10 wt% aluminum nitrate solution, shaking up, placing for 6 minutes, adding 10mL of sodium hydroxide test solution (4.3g of sodium hydroxide is dissolved in 100mL of water), adding water to the scales, shaking up, and placing for 15 minutes. And (3) taking a corresponding reagent without a reference substance as a blank, measuring the absorbance at the wavelength of 500nm by adopting an ultraviolet-visible spectrophotometry, and drawing a standard curve by taking the absorbance as a vertical coordinate and the concentration as a horizontal coordinate.

(3) And measuring the content of total flavone.

Respectively taking equal amounts of the extract of example 1 and the extract of comparative example 2, precisely weighing, placing in a Soxhlet extractor, adding chloroform, heating and refluxing for extraction until the extract is colorless, discarding chloroform solution, volatilizing chloroform from the residue, adding methanol to continue extraction until colorless (about 4 hours), evaporating the extract to dryness, dissolving the residue with diluted ethanol, transferring to a 50mL measuring flask, adding diluted ethanol (529mL ethanol is water to constant volume of 1L) to scale, and shaking up to obtain a stock solution of a sample.

Taking the stock solution of the sample, filtering, precisely measuring 5ml of the subsequent filtrate, placing in a 25ml measuring flask, adding water to dilute to the scale, and shaking up. Precisely measuring 2mL, placing in a 25mL measuring flask, adding water to 6mL, adding 5% sodium nitrite solution 1mL, shaking, standing for 6 minutes, adding 10 wt% aluminum nitrate solution 1mL, shaking, standing for 6 minutes, adding sodium hydroxide test solution (4.3g sodium hydroxide is dissolved in 100mL water) 10mL, adding water to scale, shaking, standing for 15 minutes to obtain the test solution. The absorbance of the test solution was measured at a wavelength of 500nm by ultraviolet-visible spectrophotometry.

The total flavone content is calculated by anhydrous rutin, the anhydrous rutin standard substance is prepared into standard solutions with different concentrations, the light absorption values of the standard solutions are measured, a standard curve is drawn, and the measured light absorption values of the sample solutions are substituted into a formula of the standard curve to obtain the rutin content in the sample.

(II) detection method of total polyphenol content of traditional Chinese medicine active substances

Total polyphenols were detected according to GB/T8313-.

(III) detection method of total sugar content of traditional Chinese medicine active substance

The total sugar content in the samples was determined according to the sulphuric acid phenol method.

(1) And preparing a standard curve.

0.025g of glucose standard solution is precisely weighed and is made into 500ml by deionized water. 0ml, 0.2ml, 0.4ml, 0.6ml, 0.8ml, 1.0ml, 1.2ml, 1.4ml, 1.6ml and 1.8ml are respectively sucked into a 25ml colorimetric tube, and water is added to the colorimetric tube until the volume is 2.0 ml. Precisely adding 1ml of 6 wt% phenol solution, shaking, rapidly and vertically adding 5ml of concentrated sulfuric acid with a straight pipette, shaking, and cooling for 30 min. The absorbance was measured at 490nm wavelength. And drawing a standard curve by taking the absorbance as a vertical coordinate and the concentration as a horizontal coordinate.

(2) And preparing and detecting a sample.

Weighing 0.025g of sample, placing the sample in a 100ml volumetric flask (according to the requirement of the sample, part of the sample which is not easy to dissolve can be adjusted to pH 10 by 0.1mol/L NaOH), carrying out ultrasound for 30min, filtering by a 0.45um nylon filter head, removing primary filtrate, sucking 0.2ml of filtrate into a 25ml colorimetric tube, adding water to 2.0ml, precisely adding 1ml of 6% phenol solution, shaking up, rapidly and vertically adding 5ml of concentrated sulfuric acid by a straight pipette, shaking up, cooling for 30min, and measuring absorbance at 490nm wavelength.

The total sugar content was calculated in comparison to the standard curve.

(IV) the results of the detection

The results are shown in Table 2.

TABLE 2 active ingredient content

As can be seen from the data in Table 2, the content of the active substances in the Chinese medicinal extractive solutions prepared in examples 1-3 is significantly higher than that of the Chinese medicinal extractive solution extracted in comparative example 2 by the conventional method, which indicates that the extract of JINDANKANGMIN Chinese medicinal material obtained by the extraction method of the present invention has higher active substance content.

Example 5 inhibition of anti-allergic traditional Chinese medicine extract of Jindan to inflammatory factors

(I), test materials and reagents

Raw264.7 mouse macrophage lines were purchased from the cell bank of the Chinese academy of sciences (Cat # TCM13), and CCK-8 was purchased from Dojindo (Cat # CK 04); DMEM medium, serum were purchased from Gibco (Cat #12800017, Cat # 10100147); cell 96 culture plates and common 96-well elisa plates were purchased from Corning, respectively. IL-6(Cat #88706422) cytokine ELISA kits were purchased from eBioscience, USA.

(II) RAW264.7 cell viability assay

Preparation of test samples: the jindan anti-allergy extract of example 1 was diluted with deionized water to volume concentrations of 0.313%, 0.625%, 1.25%, 2.5%, 5%, and 10% for use.

Control group: an equivalent amount of deionized water.

RAW264.7 mouse macrophage cell line in 5% CO₂Under the condition of 37 deg.C, the cells were cultured normally in 10% FBS DMEM medium, and when the cells grew to 90% density, the cells were cultured at 8X 10⁵The density of the solution/ml is inoculated in a 96-well plate, after 8 hours of adherence, test samples are added for respective treatment for 24 hours, then CCK reagent is added according to the instructions for incubation for 2 hours, and the absorbance is measured at 450 nm.

Cell viability (%) - [ a (dosed) -a (blank) ]/[ a (not dosed) -a (blank) ] × 100%

A (dosing): absorbance of the test group with cells, CCK reagent and test sample.

A (blank): absorbance of the test group with medium and CCK reagent without cells.

A (no drug addition): absorbance of test group with cells, CCK reagent, and no test sample.

As can be seen from FIG. 1, when the dilution concentration of the jindan anti-allergy extract in the system is not more than 5%, the jindan anti-allergy extract has no cytotoxicity to cells, but has a certain cell proliferation effect, which shows that the jindan anti-allergy extract of the invention can effectively promote the cell proliferation when the dilution concentration is not more than 5%. At a dilution concentration of 2.5%, the cells were most viable and had the greatest cell proliferation effect.

(III) RAW264.7 cell anti-inflammatory assay

Samples one to three: the jindan anti-allergy extract of examples 1-3 was diluted to a concentration of 2.5%;

comparative sample one: the 9 single-component extracts of comparative example 1 were each diluted to a concentration of 2.5%.

Comparative sample two: comparative example 2 the extract was diluted to a concentration of 2.5%.

Stimulus: bacterial Lipopolysaccharides (LPS).

RAW264.7 mouse macrophage cell line in 5% CO₂Under the culture condition of 37 ℃, the cells are normally cultured in a DMEM medium containing 10% FBS, and when the cells grow to 90% density, the cells are cultured according to the formula of 8X 10⁵The density of the solution/ml is inoculated on a 96-well plate, after the solution is adhered to the wall for 8 hours, the solution is randomly divided into a blank control group, a stimulation group, a positive control group, a sample group, a comparative group and a comparative group according to different added substances, wherein each group is provided with 3 multiple wells; the positive control drug with anti-inflammatory effect used was dexamethasone at a concentration of 5. mu. mol/l. Wherein:

blank control group: adding 2mL of DMEM medium;

stimulation group: 2mL of DMEM medium with 1. mu.g/mL LPS was added;

positive control group: 2mL of DMEM medium containing 1. mu.g/mL LPS and 5. mu. mol/l dexamethasone was added;

one group of samples: 1.95mL of DMEM medium containing 1. mu.g/mL LPS and 0.05mL of sample one, i.e., sample concentration 2.5%;

two groups of samples: 1.95mL of DMEM medium containing 1. mu.g/mL LPS and 0.05mL of sample two, i.e., sample concentration 2.5%;

three groups of samples: 1.95mL of DMEM medium containing 1. mu.g/mL LPS and 0.05mL of sample III, namely the sample concentration is 2.5%;

comparative example one and comparative example two: adding 1.95mL of DMEM medium containing 1 mu g/mL LPS, and 0.05mL of a first comparison sample and a second comparison sample;

after adding the drug or sample to each group, the mixture was mixed with 5% CO₂After culturing at 37 ℃ for 24h under saturated humidity, the supernatant was used for detecting interleukin-6 (IL-6) inflammatory factor, and IL-6 content was measured by ELIS kit according to the instruction procedures, and the results are shown in tables 3 and 4.

TABLE 3 results of cellular anti-inflammatory assays

TABLE 4 results of anti-inflammatory cell test of single-component extracts of group 9 of the comparative example group

As can be seen from the data in Table 3, the IL-6 inflammatory factor secretion in the blank control group was about 300pg/ml, which was less than 500 pg/ml. The secretion of IL-6 inflammatory factor in the stimulated group added with LPS was more than 3000 pg/ml. The positive control group with dexamethasone drug had an IL-6 inflammatory factor secretion of about 1800pg/ml, indicating that the stimulation model was effective.

After the traditional Chinese medicine extract prepared in the examples 1-3 of the invention is added, the secretion amount of the IL-6 inflammatory factor is reduced to about 1627pg/ml on average, which shows that the anti-inflammatory and anti-allergy effects of the traditional Chinese medicine extract under the condition of the dilution concentration of 2.5 percent are better than the anti-inflammatory and anti-allergy effects of dexamethasone with the concentration of 5 mu mol/l (the conventional administration concentration of dexamethasone).

Comparative example 2 the traditional Chinese medicine raw materials of formula 1 were extracted by a conventional method in the prior art, and the secretion of IL-6 inflammatory factor of the obtained extract at a dilution concentration of 2.5% was 2350, indicating that the extract of comparative example 2 also has a certain anti-inflammatory and anti-allergy effect. However, the average IL-6 inflammatory factor secretion of examples 1-3 of the present invention is only 69% of that of comparative example 2, which is reduced by 31%, indicating that the anti-inflammatory and anti-allergic effects of the Chinese medicinal extractive solution obtained by the extraction method of the present invention are significantly better.

The secretion amount of IL-6 inflammatory factor of the Chinese herbal medicine extract in the comparative example 1 is much higher than that of the IL-6 inflammatory factor in the examples 1-3 and the positive control group. The average IL-6 inflammatory factor secretion of examples 1 to 3 according to the invention was less than 64.3% of that of the monocomponent of comparative example 1, i.e. the IL-6 inflammatory factor secretion was reduced by more than 35.7% compared to the monocomponent extract. Compared with the single-component extract with the same amount, the Jindan anti-allergy traditional Chinese medicine extract obtained by combining the specific extraction process under the specific compound component condition of the invention has obvious synergistic effect on anti-inflammatory and anti-allergy effects. The Jindan anti-allergy traditional Chinese medicine extract has excellent anti-inflammatory and anti-allergy effects.

Example 6 repair of hormone dependent dermatitis

(I), test materials and reagents

Healthy male guinea pigs 30, 2 months old, weighed between 300 and 350g (purchased from the centre of medical laboratory animals, Guangdong province).

(II) Experimental method

An area of 3 x 3cm was selected from the back of each guinea pig for use as a test area, and hair was removed. The halometasone cream of 0.05 percent and 0.5g is externally applied and smeared twice a day until the halometasone cream is fully absorbed for 60 days. Establishing an animal model of glucocorticoid-dependent dermatitis. After the mouse is stopped for three days, the skin of the mouse is red and swollen, the mouse is itchy, and the modeling is successful.

30 guinea pigs were randomly divided into three groups. The medicine is divided into the following parts according to different medicines:

positive control group: 0.5g pimecrolimus cream; experimental groups: 1ml of the essence of example 6; combination group: 0.5g pimecrolimus cream +1ml of the essence of example 6; the preparation is administered twice daily for 21 days.

After 21 days, the weight of the guinea pig is 0-3 points according to the size of erythema, edema and pimple area on the back, respectively.

Efficacy index is (pre-treatment total score-post-treatment total score)/pre-treatment total score.

The judgment standard of the curative effect is as follows:

the curative effect index is more than or equal to 90 percent, and the healing is achieved;

the curative effect index is more than or equal to 60 percent, and the effect is obvious;

the curative effect index is more than or equal to 30 percent, which is improvement;

the efficacy index was < 30% and was not effective.

Effective rate (cure + show effect)/total number of cases x 100%

(III) results of the experiment

According to the statistical result, the effective rate of the positive control group is 62.96 percent; the effective rate of the experimental group is 82.14%, and the effective rate of the combined group is 92.85%. Compared with a positive control group, the effective rate of the test group is improved by 30.5 percent. The essence prepared from the gold lead anti-allergy traditional Chinese medicine extract can effectively repair hormone-dependent dermatitis, has an effect obviously better than that of the existing hormone medicine halometasone cream for treating dermatitis, and has no side effect caused by hormone medicines. When the essence prepared from the Jindan anti-allergy traditional Chinese medicine extract is used together with the immunosuppressive drug pimecrolimus cream for treating dermatitis, the effective rate for treating dermatitis is the highest. Therefore, the combination of the essence prepared from the Jindan anti-allergy traditional Chinese medicine extract and the pimecrolimus cream is a good choice for repairing hormone-dependent dermatitis. The Jindan anti-allergy traditional Chinese medicine extract can be used for preparing an anti-allergy ointment.

Example 7: clinical examination

(I) preparing the face skin care product for testing

Cleansing milk: 5 parts of traditional Chinese medicine extracting solution, 10 parts of tridecyl alcohol polyether sodium sulfate, 5 parts of sodium lauroamphoacetate, 2 parts of coco glucoside, 1 part of coco alcohol, 0.5 part of PEG-120 methyl glucose dioleate, 0.2 part of carbomer, 0.2 part of sodium hydroxide, pH of citric acid adjusted to 6-7, a proper amount of preservative and water added to 100 parts.

The preparation method of the facial cleanser comprises the following steps:

(1) the formula amounts of sodium trideceth sulfate, sodium lauroamphoacetate, cocoglucoside, cocoalcohol, PEG-120 methyl glucose dioleate and 30 parts of water were heated to 80 ℃ and dispersed with stirring until clear.

(2) And (2) soaking carbomer in the residual water until the carbomer is wet, dispersing until the carbomer is uniform and transparent, adding the reaction system obtained in the step (1), stirring, cooling to 45 ℃, adding the residual components, and stirring uniformly.

Essence: 10 parts of traditional Chinese medicine extracting solution, 2 parts of glycerol, 1.5 parts of dipropylene glycol, 1 part of betaine, 1 part of phenyl trimethicone, 160.5 parts of octyl dodecanol polyether, 0.5 part of glycerol acrylate/acrylic acid copolymer, 0.2 part of carbomer, 0.2 part of triethanolamine, 0.6 part of sodium hyaluronate and 100 parts of water.

The preparation method of the essence comprises the following steps: mixing the glycerol and the dipropylene glycol with the sodium hyaluronate uniformly in advance according to the formula ratio, adding water to soak the carbomer, and heating and stirring the carbomer uniformly; then cooling to 45 ℃, adding the rest components, and stirring uniformly.

(II) test method

Selecting 30 volunteers, wherein the skin types are: dry skin 15 people, neutral skin 6 people, oily skin 9 people, total 30 people. According to the skin problem distribution of the target population: easily allergic 14 people and easily pruritus 16 people.

After the target people clean the face with the facial cleanser every night, the face is massaged by using the essence until the essence is absorbed, and the facial cleanser is continuously used for 4 weeks.

Evaluation of skin red and swelling and pruritus problems:

the effect is shown: after the 4-week test, the skin on the cheek area (1 h after washing) was significantly more red and itchy than the initial one.

The method has the following advantages: after the 4-week trial, the cheek area skin (1 h after washing) was improved in redness and itching compared to the initial.

And (4) invalidation: after the 4-week trial, the cheek area skin (1 h after washing) was comparable or worse in the red, swollen and itchy state than initially.

(III) test results

And (3) test results: the skin red swelling and pruritus problems of 30 people are improved, and statistical analysis shows that 78.7 percent of the skin red swelling and pruritus problems are obvious in effect, 18.2 percent of the skin red swelling and pruritus problems are effective, and the total effective rate is 96.9 percent. The local skin change (1 h after washing) of the face at the initial, 2 and 4 weeks of use in 1 volunteer with red, swollen and itching face is shown in fig. 2. The local skin changes (1 h after washing) of the face at the initial, 2 and 4 weeks of use in 1 volunteer with severe redness and itching due to hormonal face are shown in fig. 3.

As can be seen from FIG. 2, the volunteer had flaky, red and swollen facial skin at the beginning of use, and severe red, swollen and itching; after 2 weeks of use, the skin red swelling and pruritus are obviously improved, and the red swelling is almost eliminated; after 4 weeks of use, the skin surface returned to normal without redness and swelling.

As can be seen from FIG. 3, the volunteer had flaky, red and swollen facial skin at the beginning of use, severe red and swollen itching, dry desquamation; after 2 weeks of use, the red swelling and pruritus of the skin are obviously improved, the red swelling is almost eliminated, the desquamation phenomenon is obviously reduced, and the skin looks more moist and glossy; after 4 weeks of application, the skin surface recovered normal without redness and swelling and without desquamation.

The test results show that the cosmetics prepared from the jindan anti-allergy traditional Chinese medicine extract can play a role in resisting inflammation and allergy, and can effectively improve the red swelling and pruritus of the skin and repair hormone-dependent dermatitis.

The embodiments of the present invention have been described in detail, but the embodiments are merely examples, and the present invention is not limited to the embodiments described above. Any equivalent modifications or alterations to this practice will occur to those skilled in the art and are intended to be within the scope of this invention. Accordingly, equivalent changes and modifications made without departing from the spirit and scope of the present invention should be covered by the present invention.

Claims

1. The preparation method of the jindan anti-allergy traditional Chinese medicine extract is characterized by comprising the following steps:

2. The preparation method of the jindan anti-allergy traditional Chinese medicine extract as claimed in claim 1, wherein in the step (1), the raw material medicines in parts by weight comprise: 20-25 parts of scutellaria baicalensis, 10-14 parts of radix sophorae flavescentis, 5-10 parts of coptis chinensis, 3-5 parts of phellodendron amurense, 10-15 parts of rhizoma smilacis glabrae, 15-18 parts of honeysuckle, 8-13 parts of peony root bark, 6-10 parts of fructus cnidii and 5-8 parts of fructus kochiae.

3. The preparation method of the jindan anti-allergy traditional Chinese medicine extract as claimed in claim 2, wherein in the step (1), the raw material medicines in parts by weight comprise: 20 parts of scutellaria baicalensis, 14 parts of sophora flavescens, 8 parts of coptis chinensis, 4 parts of phellodendron amurense, 12 parts of rhizoma smilacis glabrae, 18 parts of honeysuckle, 13 parts of peony root bark, 6 parts of fructus cnidii and 5 parts of fructus kochiae.

4. The method for preparing the jindan anti-allergy Chinese medicinal extract according to claim 1, wherein in the step (1), the solvent is water or an aqueous ethanol solution; the mass ratio of the raw material medicine to the solvent is 1 (10-50), and the raw material medicine is soaked for 1-3 hours.

5. The method for preparing the jindan anti-allergy traditional Chinese medicine extract as claimed in claim 1, wherein in the low-temperature ultrasonic extraction step, the ultrasonic temperature is as follows: 25-50 ℃, the ultrasonic frequency is 20-50 kHz, and the single ultrasonic time is as follows: 15 min-30 min; the ultrasonic frequency is as follows: 3 times to 5 times.

6. The method for preparing the jindan anti-allergy traditional Chinese medicine extract as claimed in claim 1, wherein the microfiltration membrane filtration step adopts a 0.1-1 μm microfiltration membrane for filtration to obtain a filtrate L; and in the reverse osmosis membrane pressurization concentration step, the filtrate L is concentrated to 20-50% of the volume of the filtrate L through a reverse osmosis membrane under 0.5-3 MPa.

7. The jindan anti-allergy Chinese medicinal extract obtained by the preparation method of the jindan anti-allergy Chinese medicinal extract as claimed in any one of claims 1 to 6.

8. The use of the jindan anti-allergy traditional Chinese medicine extract as claimed in claim 7, which is used for preparing an anti-allergy cosmetic or an anti-allergy ointment.

9. The application of the jindan anti-allergy traditional Chinese medicine extract as claimed in claim 8, wherein the anti-allergy cosmetic or the anti-allergy ointment comprises the jindan anti-allergy traditional Chinese medicine extract and an auxiliary material, the content of the jindan anti-allergy traditional Chinese medicine extract is 1-40 wt%, and the content of the auxiliary material is 60-99 wt%.

10. The use of the jindan anti-allergy traditional Chinese medicine extract as claimed in claim 8, wherein the anti-allergy cosmetic comprises facial cleanser, face cleanser, facial mask, facial cream, essence water, toner and smoothing toner.