CN113262179A

CN113262179A - Plant extract composition with anti-allergy effect and preparation method and application thereof

Info

Publication number: CN113262179A
Application number: CN202110296398.8A
Authority: CN
Inventors: 李远彬; 张嘉恒; 徐美禄; 丘彩妮; 陈一凡; 王振元
Original assignee: Shenzhen Xuanjia Biological Technology Co ltd
Current assignee: Shenzhen Xuanjia Biological Technology Co ltd
Priority date: 2021-03-19
Filing date: 2021-03-19
Publication date: 2021-08-17

Abstract

The invention relates to the technical field of cosmetics, and discloses a plant extract composition with an anti-allergy effect and a preparation method thereof, wherein the composition comprises extracts of tetrandra root, gentian and liquorice; the content of each plant extract is as follows according to weight percentage: 1 to 50 percent of tetrandra root, 1 to 50 percent of gentian and 1 to 50 percent of liquorice; the composition compounded by the plant extracts can inhibit allergic reaction and reduce inflammation from various ways, and the composition can more effectively exert the anti-allergy effect; the plant composition with the anti-allergy effect is extracted by taking the deep eutectic solvent as the extraction solvent, and has the characteristic of higher extraction efficiency than the traditional water extraction method or the simple water-alcohol combined extraction method, and the anti-allergy effect of the extract is more remarkable.

Description

Plant extract composition with anti-allergy effect and preparation method and application thereof

Technical Field

The invention relates to the technical field of cosmetics, in particular to an anti-allergy plant composition, a preparation method and application thereof.

Background

In recent years, with the change of human dietary structure and the increase of life pressure, air pollution is increased, people with skin allergy are increased rapidly, and many visitors in the world feel that the people are sensitive skin. Sensitive skin is mostly present on the face, and other parts of the body such as hands, feet, back, etc. may be present. Especially asian people are more prone to have sensitive skin problems. Sensitive skin is mainly characterized by crisp, thin, dry and desquamation of skin, dry, tight, burning, stabbing pain of skin, skin itch, erythema and erythra.

The method for treating sensitive skin in medicine mainly comprises the step of treating with drugs such as glucocorticoid and histamine receptor antagonist, wherein the drugs can effectively relieve skin allergy or irritation, but after long-term repeated use, the anti-skin allergy drugs can generate obvious side effects, such as skin atrophy, telangiectasia, acneitin abnormality and the like, the glucocorticoid can form dependence after long-term use, skin problems can be aggravated after drug withdrawal, even the immunity of the organism is reduced, and the histamine receptor antagonist can cause central nervous system inhibition. Therefore, the drug therapy is not suitable for daily care and use of sensitive skin, and some skin care products of anti-allergen materials which are safe and effective to use and have no side effect are popular with people with sensitive skin.

In recent years, the cosmetic industry is developing towards green, natural and pure directions, so that natural plant extracts with excellent anti-allergy effects are developed and applied to anti-allergy cosmetics, and the anti-allergy cosmetic has urgent needs and wide market prospects.

Disclosure of Invention

The application aims to provide a plant extract composition with anti-allergy effect, a preparation method and application thereof, and the composition has reliable effect and has the effects of inhibiting hyaluronidase and resisting histamine.

In a first aspect, the present application provides a plant extract composition having anti-allergy effects, the composition comprising extracts of tetrandra root, gentian root and licorice root; the content of each plant extract is as follows according to weight percentage: 1 to 50 percent of tetrandra root, 1 to 50 percent of gentian root and 1 to 50 percent of liquorice.

It should be noted that the percentage of each plant extract in the present application is based on the dry weight of the composition, but that solutions containing the composition are often obtained during the preparation process.

In a second aspect, a method for preparing a plant extract composition with anti-allergy effect comprises the following steps:

respectively weighing betaine, 1, 3-butanediol and 1, 2-hexanediol according to the molar ratio of the betaine, the 1, 3-butanediol and the 1, 2-hexanediol of (1-5) to (1-5) and stirring uniformly at the temperature of 20-60 ℃ until the mixture is colorless and transparent to obtain a deep eutectic solvent, and adding a certain amount of water to prepare a deep eutectic solvent water solution, wherein the volume ratio of the deep eutectic solvent water solution is 10-95%;

weighing radix stephaniae tetrandrae, radix gentianae and liquorice, wherein the contents of the radix stephaniae tetrandrae, the radix gentianae and the liquorice are as follows in percentage by weight: 1-50% of tetrandra root, 1-50% of gentian and 1-50% of liquorice, and soaking the tetrandra root, the gentian and the liquorice which are measured by using the obtained deep eutectic solvent aqueous solution at the temperature of 30-80 ℃ to obtain an extracting solution;

performing microfiltration clarification on the extracting solution by using a ceramic membrane with the aperture of 100-500nm, intercepting insoluble particles and large molecules, and collecting clarified permeate;

concentrating the permeate, and loading to C18 or C8 reversed phase chromatographic column with filler particle size of 5-60 μm; the inner diameter of the chromatographic column is 10-1000 mm. The mobile phase is ethanol water or methanol water, gradient or step elution is adopted, and target substance fractions are collected and merged according to the liquid phase detection result;

concentrating the target fraction, adding 1, 3-butanediol, stirring, standing for 12-36 hr, and collecting supernatant;

centrifuging the supernatant to obtain a centrifuged supernatant, wherein the centrifuged supernatant contains the plant extract composition with anti-allergy effect.

Preferably, the tetrandra root, the gentian and the liquorice are measured, and the content of the tetrandra root, the gentian and the liquorice is as follows according to the weight percentage: 1-50% of stephania tetrandra, 1-50% of gentian and 1-50% of liquorice, and soaking the stephania tetrandra, the gentian and the liquorice which are measured by using the obtained deep eutectic solvent aqueous solution at the temperature of 30-80 ℃ to obtain an extracting solution, wherein the extracting solution specifically comprises:

weighing radix stephaniae tetrandrae, radix gentianae and liquorice, wherein the contents of the radix stephaniae tetrandrae, the radix gentianae and the liquorice are as follows in percentage by weight: 1-50% of tetrandra root, 1-50% of gentian and 1-50% of liquorice, wherein the total weight of the tetrandra root, the gentian and the liquorice is the numerical value A by taking gram as a unit;

recording the volume of the deep eutectic solvent aqueous solution as a value B by taking milliliters as a unit, and measuring the prepared deep eutectic solvent aqueous solution to ensure that the value B is 5-20 times of the value A;

soaking and extracting the stephania tetrandra, the gentian and the liquorice in 10-95% of deep eutectic solvent aqueous solution for 1-3 times, wherein each soaking and extracting time is 2-48 hours, and combining the extracting solutions.

Preferably, the mobile phase is an ethanol aqueous solution, wherein the volume ratio of the ethanol aqueous solution is 10-95%.

Preferably, in the step of concentrating the target fraction, adding 1, 3-butanediol, stirring uniformly, standing for 12-36 hours, and collecting the supernatant, the target fraction is concentrated by 10-20 times.

Preferably, in the step of concentrating the target fraction, adding 1, 3-butanediol, stirring uniformly, standing for 12-36 hours, and collecting the supernatant, the addition amount of 1, 3-butanediol is 1-10 times of the volume of the eluate.

In a third aspect, the present application provides the use of a plant extract composition as described above for the preparation of a skin care cosmetic or pharmaceutical product, including creams, lotions, gels, masks and lotions.

Compared with the prior art, the method has the following beneficial effects:

1. the alkaloid in the tetrandra root and the iridoid glycoside in the gentiana have the effect of a corticoid-like medicament, and the alkaloid and the iridoid glycoside in the gentiana root are not only allergic mediator antagonists but also allergic mediator blockers, so that the composition has reliable resisting and relieving effects on allergic reactions. The terpenoid component in Glycyrrhrizae radix has effects of regulating immunity and inhibiting complement, enhancing immunity and inhibiting allergy. The composition compounded by the plant extracts can inhibit allergic reaction and reduce inflammation from various ways, so that the composition can more effectively exert the anti-allergy effect;

2. the deep eutectic solvent is prepared by combining betaine, 1, 3-butanediol and 1, 2-hexanediol according to a specific proportion, and is a common safe component for cosmetics. Betaine chemistry is a commonly used humectant for cosmetics, is very safe, unlimited in use, and little in skin irritation, and serves as a hydrogen bond receptor in a eutectic solvent. 1, 3-butanediol and 1, 2-hexanediol which are used as hydrogen bond donors are all common ingredients of cosmetics, and the using amount is not limited at all, wherein the 1, 2-hexanediol has the amphipathy of lipophilicity and hydrophilicity. The invention can simultaneously extract active matters with different hydrophilic and lipophilic characteristics in the anti-allergic plant composition by the specially combined deep eutectic solvent. The plant composition with the anti-allergy effect is extracted by taking the deep eutectic solvent as the extraction solvent, has the characteristic of higher extraction efficiency than the traditional water extraction method or the simple water-alcohol combined extraction method, and has more remarkable anti-allergy effect.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Example 1

Weighing 10g of tetrandra root, 30g of gentian and 20g of liquorice, and soaking and extracting the tetrandra root, the gentian and the liquorice in 20 times of deep eutectic solvent aqueous solution (the molar ratio is 1:2:2) containing 50% of betaine-1, 3-butanediol-1, 2-hexanediol (the molar ratio is 1:2:2) at 30 ℃ for 1 time, and soaking for 2 hours each time to obtain an extracting solution;

performing microfiltration and clarification on the extracting solution by using a ceramic membrane with the aperture of 500nm to obtain a clarified solution;

the supernatant was concentrated 1-fold and applied to a C18 reverse phase column (25X 250mm,10 μm). The mobile phase is ethanol water, the flow rate is 20mL/min, and gradient elution is adopted; 0-15min, 10% ethanol; 15-25min, 20% ethanol; 25-35min, 50% ethanol, 35-65min, 100% ethanol. The detection wavelength is 237nm, and the target components are collected in sections according to the liquid phase detection result.

Concentrating the target fraction by 10 times, adding 1, 3-butanediol with the volume of 2 times, uniformly stirring, standing for 24 hours, and collecting the supernatant;

centrifuging the supernatant to obtain centrifugal supernatant, wherein the centrifugal supernatant contains the plant extract composition with anti-allergy effect.

Example 2

Weighing 5g of tetrandra root, 40g of gentian and 10g of liquorice, soaking and extracting for 2 times at 80 ℃ by using 10 times (volume of solution (ml): weight of medicinal material) of deep eutectic solvent aqueous solution containing 70% of betaine-1, 3-butanediol-1, 2-hexanediol (molar ratio is 1:3:1) for 1 hour each time to obtain an extracting solution;

performing microfiltration and clarification on the extracting solution by using a ceramic membrane with the aperture of 100nm to obtain a clarified solution;

the supernatant was concentrated 2-fold and applied to a C18 reverse phase column (25X 250mm,10 μm). The mobile phase is methanol water, the flow rate is 20mL/min, and gradient elution is adopted; 0-15min, 10% methanol; 15-25min, 20% methanol; 25-35min, 50% methanol, 35-65min, 100% methanol. The detection wavelength is 237nm, and the target components are collected in sections according to the liquid phase detection result.

Concentrating the target fraction by 5 times, adding 1 time volume of 1, 3-butanediol, stirring uniformly, standing for 12 hours, and collecting the supernatant;

Example 3

Weighing 40g of tetrandra root, 10g of gentian and 20g of liquorice, soaking and extracting for 3 times at 40 ℃ by using 30 times (volume of solution (ml): weight of medicinal materials) of deep eutectic solvent aqueous solution containing 30% of betaine-1, 3-butanediol-1, 2-hexanediol (molar ratio is 1:4:2) for 1.5 hours each time to obtain an extracting solution;

performing microfiltration clarification on the extracting solution by using a ceramic membrane with the aperture of 200nm to obtain a clarified solution;

the supernatant was concentrated 5-fold and applied to a C18 reverse phase column (25X 250mm,10 μm). The mobile phase is methanol water, the flow rate is 20mL/min, and gradient elution is adopted; 0-15min, 10% methanol; 15-25min, 20% methanol; 25-35min, 50% methanol, 35-65min, 100% methanol. The detection wavelength is 237nm, and the target components are collected in sections according to the liquid phase detection result.

Concentrating the target fraction by 8 times, adding 1, 3-butanediol with the volume of 3 times, uniformly stirring, standing for 12 hours, and collecting the supernatant;

Example 4

Soaking 30g of radix Stephaniae Japonicae, 30g of radix Gentianae and 30g of Glycyrrhrizae radix in 15 times (volume of solution (ml): weight of medicinal material) of deep eutectic solvent aqueous solution containing 90% betaine-1, 3-butanediol-1, 2-hexanediol (molar ratio of 2:3:2) at 50 deg.C for 3 times (each time for 2 hr) to obtain extractive solution;

the supernatant was concentrated 2-fold and applied to a C18 reverse phase column (25X 250mm,10 μm). The mobile phase is ethanol water, the flow rate is 20mL/min, and gradient elution is adopted; 0-15min, 10% ethanol; 15-25min, 20% ethanol; 25-35min, 50% ethanol, 35-65min, 100% ethanol. The detection wavelength is 237nm, and the target components are collected in sections according to the liquid phase detection result.

Concentrating the target fraction by 20 times, adding 1, 3-butanediol with the volume of 8 times, uniformly stirring, standing for 12 hours, and collecting supernatant;

Example 5

Taking 20g of tetrandra root, 40g of gentian and 40g of liquorice, soaking and extracting for 3 times at 60 ℃ by using 10 times (volume of solution (ml): weight of medicinal materials) of deep eutectic solvent aqueous solution containing 20% of betaine-1, 3-butanediol-1, 2-hexanediol (molar ratio is 2:3:3) for 3 hours each time to obtain extract;

the supernatant was concentrated 5-fold and applied to a C18 reverse phase column (25X 250mm,10 μm). The mobile phase is ethanol water, the flow rate is 20mL/min, and gradient elution is adopted; 0-15min, 10% ethanol; 15-25min, 20% ethanol; 25-35min, 50% ethanol, 35-65min, 100% ethanol. The detection wavelength is 237nm, and the target components are collected in sections according to the liquid phase detection result.

Concentrating the target fraction by 10 times, adding 1, 3-butanediol with the volume of 5 times, uniformly stirring, standing for 36 hours, and collecting supernatant;

Application example 1

The plant extract composition having whitening effects, which is prepared in examples 1 to 5, is prepared into skin care cosmetics or medicines including creams, lotions, gels, masks, embrocations and lotions.

In order to demonstrate the effect of the anti-allergy composition provided by the present invention, an efficacy experiment of the anti-allergy composition provided by the present invention is given below.

The anti-allergy composition provided in embodiments 1 to 5 of the present invention is added to a skin care product matrix, and the mixture is mixed and recorded as sample 1, sample 2, sample 3, sample 4, and sample 5 in sequence, wherein the skin care product matrix is a skin care product matrix conventional in the art, and the ratio of the anti-allergy composition to the skin care product matrix is as follows: 2:98,

1. an anti-allergy efficacy test method and test results;

(1) the test method briefly describes: hyaluronidase inhibition assay

Hyaluronidase is a implicated enzyme in type i allergic reactions and can affect the release of histamine from mast cells. The higher the activity of hyaluronidase, the more histamine is released by mast cells. Therefore, the activity of inhibiting the hyaluronidase is effective in resisting allergy. The hyaluronidase can react with sodium hyaluronate to generate beta-N-glucosamine, the substance reacts with acetylacetone under alkaline conditions to generate chromogen 2-methyl-3-diacetylpyrrole derivatives, the chromogen and an Ellisib reagent generate pink under the condition of concentrated hydrochloric acid and ethanol, scanning is carried out at the wavelength of 530nm to obtain a light absorption value, and the inhibition of a test substance on the hyaluronidase can be obtained by utilizing the following formula.

Hyaluronidase inhibition (%) ((A-B) - (C-D))/(A-B) × 100%

A: OD value measured with control solution (deionized water instead of sample solution)

B: OD value measured for control blank solution (deionized water instead of test substance, and acetate buffer instead of hyaluronidase)

C: OD value measured for test substance

D: OD value of blank sample (acetate buffer instead of hyaluronidase)

(2) And (4) result judgment standard:

TABLE 2-1 evaluation criteria for antiallergic efficacy

Evaluation standard of serial number antiallergic effect

1 the inhibition rate of ineffective hyaluronidase is less than 20 percent

2, the inhibition rate of the low-efficiency hyaluronidase is 20 to 50 percent

3 the inhibition rate of the middle-effect hyaluronidase is 50 to 75 percent

4 the high-efficiency hyaluronidase inhibition rate is more than 75 percent

(3) And (3) testing results: the specific test results are shown in table 1.

TABLE 1 results of the anti-allergic efficacy testing of the samples

Serial number	Sample name	Hyaluronidase inhibition (%)	Antiallergic effect
				1	Sample 1	86.7	Efficient
2	Sample 2	83.1	Efficient
				3	Sample 3	88.6	Efficient
4	Sample No. 4	95.7	Efficient
				5	Sample No. 5	92.1	Efficient
6	Substrate	0	Invalidation
				7	A certain commercial product	51.6	Middle effect

2. Histamine resistance test method and test results

(1) Brief description of the test methods

80 white guinea pigs with the weight of 250-: the sample group and the triamcinolone acetonide acetate urea soft paste group are 10 in each group, and the male and female parts are respectively half. The skin of the area where the hair was removed on the back of the right hind paw was rubbed gently with fine sandpaper to make it red but not bleed, then 0.2mL of the test drug was applied to the wound surface at an area of about 1.0cm2, and 0.03mL of histamine was applied to the back of each guinea pig foot from low to high concentrations at intervals of 3min, the histamine concentrations being 1.0X 10-4, 1.5X 10-4, 2.0X 10-4, 2.5X 10-4, 3.0X 10-4, 3.5X 10-4, and 4.0X 10-4, respectively. When the guinea pigs have a pruritic response, i.e., licking the right hind paw wound site (or until the highest concentration has been applied and no pruritic response has occurred), the amount of histamine administered to each guinea pig is accumulated, i.e., the amount of histamine tolerated by that guinea pig.

(2) The specific test results are shown in table 2.

TABLE 2 Histamine test results

Serial number	Sample name	Histamine tolerance (μ g)
			Positive control group	Triamcinolone acetonide acetate urea ointment group	28.63±11.85
1	Sample 1	21.63±10.25
			2	Sample 2	19.91±9.43
3	Sample 3	22.54±8.71
			4	Sample No. 4	23.65±10.15
5	Sample No. 5	24.17±11.16
			6	Substrate	10.65±7.32
7	A certain commercial product	18.68±8.21

Test results show that the antiallergic composition provided by the invention can obviously enhance the tolerance capability of guinea pig histamine, and the effect is close to hormone.

Stephania tetrandra, being cold in nature, can dispel wind, alleviate pain, induce diuresis to alleviate edema, and is used for treating rheumatic arthralgia, edema, beriberi, difficult urination, eczema and sore toxicity. Radix Stephaniae Tetrandrae has effects of resisting allergy, relieving itching, diminishing inflammation, relieving pain, subsiding swelling, and regulating immunity.

Gentiana scabra Bunge, being cold in nature, can purge excess fire of liver and gallbladder and remove damp-heat in lower-jiao, and is used for damp-heat yellow carbuncle, swelling of yin, pruritus vulvae, leukorrhagia, eczema and pruritus. Radix Gentianae has antiinflammatory, analgesic, liver protecting, gallbladder function promoting, cancer cell inhibiting, antioxidant, and antibacterial effects.

The liquorice is neutral in nature, and can tonify spleen and qi, clear heat and remove toxicity, eliminate phlegm and relieve cough, relieve spasm and pain and harmonize the drugs. The Glycyrrhrizae radix has effects of removing toxic substance, resisting inflammation, resisting allergy, eliminating phlegm, relieving cough, resisting liver injury, and resisting cancer.

The plants are traditionally eaten or used as medicines, have the functions of resisting inflammation, resisting allergy, easing pain and reducing swelling, are combined synergistically, achieve the anti-allergy effect from various ways, are synergistic synergistically, and are particularly suitable for nursing and applying sensitive skin. Meanwhile, the plants have water-soluble and oil-soluble active ingredients, and the traditional extraction method cannot extract the active ingredients at one time, so that the problem can be well solved.

Deep Eutectic Solvents (DES) are Eutectic compounds in which two or three components are connected to each other by hydrogen bond acceptors and hydrogen bond donors through hydrogen bond forces, and the hydrogen bond donors of the Deep Eutectic Solvents form hydrogen bonds with the hydrogen bond acceptors, and the freezing points of the Deep Eutectic Solvents are significantly lower than the melting points of the components when used alone. The deep eutectic solvent extraction has the advantages that water-soluble active ingredients in the plants, such as plant polysaccharide, organic acids and alkaloids, can be extracted at one time, and oil-soluble flavonoids and saponins can also be extracted. In view of the complexity of the plant components, the deep eutectic solvent is more beneficial to the extraction of the plant active components.

The application has the following beneficial effects:

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.

Claims

1. A plant extract composition with anti-allergy effect is characterized in that the composition comprises extracts of tetrandra root, gentian root and licorice; the content of each plant extract is as follows according to weight percentage: 1 to 50 percent of tetrandra root, 1 to 50 percent of gentian root and 1 to 50 percent of liquorice.

2. A preparation method of a plant extract composition with anti-allergy effect is characterized by comprising the following steps:

respectively weighing betaine, 1, 3-butanediol and 1, 2-hexanediol according to the molar ratio of the betaine, the 1, 3-butanediol and the 1, 2-hexanediol of (1-5) to (1-5) and stirring uniformly at the temperature of 20-60 ℃ until the mixture is colorless and transparent to obtain a deep eutectic solvent, and adding a certain amount of water to prepare a deep eutectic solvent aqueous solution, wherein the volume ratio of the deep eutectic solvent aqueous solution is 10-95%;

performing microfiltration clarification on the extracting solution by using a ceramic membrane with the aperture of 100-500nm, intercepting insoluble particles and macromolecules, and collecting clarified permeate;

concentrating the permeate, and loading to C18 or C8 reversed phase chromatographic column with filler particle size of 5-60 μm; the inner diameter of the chromatographic column is 10-1000 mm. The mobile phase is ethanol water or methanol water, gradient or step elution is adopted, and target fraction is collected and merged according to the liquid phase detection result;

3. The preparation method of the plant extract composition with the anti-allergy effect according to claim 2, wherein the content of the tetrandra root, the gentian root and the liquorice are as follows in percentage by weight: 1-50% of stephania tetrandra, 1-50% of gentian and 1-50% of liquorice, and soaking the stephania tetrandra, the gentian and the liquorice which are measured by using the obtained deep eutectic solvent aqueous solution at the temperature of 30-80 ℃ to obtain an extracting solution, wherein the extracting solution specifically comprises:

weighing radix stephaniae tetrandrae, radix gentianae and liquorice, wherein the contents of the radix stephaniae tetrandrae, the radix gentianae and the liquorice are as follows in percentage by weight: 1-50% of tetrandra root, 1-50% of gentian and 1-50% of liquorice, wherein the total weight of the tetrandra root, the gentian and the liquorice is the numerical value A by taking grams as a unit;

recording the volume of the deep eutectic solvent aqueous solution as a value B in milliliters, and measuring the prepared deep eutectic solvent aqueous solution to ensure that the value B is 5-20 times of the value A;

soaking and extracting the stephania tetrandra, the gentian and the liquorice in 10-95% of deep eutectic solvent aqueous solution for 2-48 hours for 1-3 times, and combining the extracting solutions.

4. The method for preparing the plant extract composition with anti-allergy effect according to claim 2, wherein the mobile phase is an ethanol aqueous solution, wherein the volume ratio of the ethanol aqueous solution is 10-95%.

5. The method of claim 2, wherein the concentration of the target substance fraction is 10 to 20 times in the step of adding 1, 3-butanediol to the concentrated target substance fraction, stirring the mixture uniformly, standing the mixture for 12 to 36 hours, and collecting the supernatant.

6. The method of claim 2, wherein 1, 3-butanediol is added to the concentrated target fraction, the mixture is stirred uniformly, the mixture is left for 12 to 36 hours, and the supernatant is collected, wherein the amount of 1, 3-butanediol added is 1 to 10 times the volume of the eluate.

7. Use of the plant extract composition according to claim 1 for the preparation of skin care cosmetics or pharmaceuticals, including creams, lotions, gels, masks and lotions.