CN113712858B - Preparation and application of antibacterial and anti-inflammatory plant extract composition - Google Patents

Preparation and application of antibacterial and anti-inflammatory plant extract composition Download PDF

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CN113712858B
CN113712858B CN202110771103.8A CN202110771103A CN113712858B CN 113712858 B CN113712858 B CN 113712858B CN 202110771103 A CN202110771103 A CN 202110771103A CN 113712858 B CN113712858 B CN 113712858B
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inflammatory
chlorogenic acid
antibacterial
mass ratio
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CN113712858A (en
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曹茜
周秋娜
金荣熙
姚清
申彦晟
金延埈
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Cosmax China Cosmetics Co Ltd
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K8/00Cosmetics or similar toiletry preparations
    • A61K8/18Cosmetics or similar toiletry preparations characterised by the composition
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    • A61K8/37Esters of carboxylic acids
    • A61K8/375Esters of carboxylic acids the alcohol moiety containing more than one hydroxy group
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00Cosmetics or similar toiletry preparations
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    • A61K8/96Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
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    • A61P29/00Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61Q17/00Barrier preparations; Preparations brought into direct contact with the skin for affording protection against external influences, e.g. sunlight, X-rays or other harmful rays, corrosive materials, bacteria or insect stings
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61QSPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
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    • A61K2800/80Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
    • A61K2800/82Preparation or application process involves sonication or ultrasonication
    • YGENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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Abstract

The invention discloses a bacteriostatic and anti-inflammatory plant extract composition and application thereof. According to the invention, the antibacterial property and the additive amount of the composition are tested by taking the compound proportion of the basil, the chlorogenic acid and the medicinal rheum officinale as the cut-in points, and finally the plant extract composition with small additive amount and strong antibacterial and anti-inflammatory effects is obtained. Wherein, the basil: chlorogenic acid: the mass ratio of the rhubarb raw materials is (1-3), 0.05-0.2 and (1-3). The composition can effectively inhibit the breeding of skin pathogenic bacteria, and reduce inflammatory reaction by reducing active oxygen free radicals, removing inflammatory factors, improving the water loss of the horny layer and the like.

Description

Preparation and application of antibacterial and anti-inflammatory plant extract composition
Technical Field
The invention belongs to the field of cosmetics, and relates to preparation and application of a bacteriostatic and anti-inflammatory plant extract composition.
Background
Due to the continuous improvement of life style of people, the continuous change of life environment, the influence of age, psychological pressure, environmental pollution, four seasons change, chemical abuse and the like, numerous injuries are brought to skin and skin mucous membrane, and the occurrence rate of skin diseases is in a continuous rising trend. Modern medical research shows that the dysbacteriosis of human body and the breeding of pathogenic bacteria can lead to endocrine dysfunction, the reaction often causes vigorous sebum secretion on the skin, and simultaneously, the skin is complicated with inflammation, acne, pimple at the top, and the like.
Ocimum basilicum is a plant of Labiatae, ocimum, herba Achilleae, herba Pogostemonis, herba Cymbidis, and herba West king, and is a medicinal and edible aromatic plant. The flavor is similar to fennel, the whole plant is small and exquisite, the leaf color is emerald green, the color is bright, and the fragrance is overflowed. Ocimum basilicum is one of the main aromatic oil plants, and the volatile oil mainly exists in stems, leaves and flowers and ears of the Ocimum basilicum, and generally contains 0.1-0.12% of oil. The whole herb of Ocimum basilicum can be used as medicine, has the property of Wen Weixin, has various effects of dispelling wind and promoting qi, activating blood, resolving dampness, promoting digestion, removing toxin and the like, and has a sterilization rate of the pure Ocimum basilicum essential oil of hundred ninety-nine percent.
Chlorogenic acid (Chlorogenic acid) is a phenylpropionic acid substance, is an ester formed by caffeic acid and quinic acid, exists in common Chinese herbal medicines such as capillary artemisia, honeysuckle and the like, has a strong broad-spectrum antibacterial effect, and the research on the biological activity of Chlorogenic acid in modern science is deep into a plurality of fields such as food, health care, medicine, daily chemical industry and the like. Chlorogenic acid is an important bioactive substance, and has antibacterial, antiviral, leukocyte increasing, liver protecting, gallbladder function promoting, antitumor, blood pressure lowering, blood lipid reducing, free radical scavenging, and central nervous system exciting effects.
Rheum officinale (Rheum palmatum L.), rheum officinale (Rheum offcinale baill.), rheum tanguticum (Rheum tanguticum Maxim. Ex Balf.), and Rheum tanguticum (Rheum tanguticum Maxim.) are generally dry roots and rhizomes of Polygonaceae. The main functions are as follows: purging heat toxin, removing accumulation and stagnation, promoting blood stasis, etc. The anthraquinone compounds include emodin, physcion, aloe-emodin, rhein, chrysophanol, etc. The rheum officinale has a broad antibacterial spectrum, and has an inhibiting effect on a plurality of gram-positive bacteria and negative bacteria such as staphylococcus aureus, hemolytic streptococcus, typhoid bacillus, shigella dysenteriae, helicobacter pylori and the like; has inhibiting effect on various fungi, influenza virus, hepatitis virus and human immunodeficiency virus.
At present, the application of the three components of the basil, the chlorogenic acid and the rheum officinale is mainly concentrated in the fields of medicines, foods and health products, and the three components are not much applied in the field of cosmetics. The Singh S finds that the basil volatile oil has obvious inhibition effect on plantar swelling caused by carrageenan of rats and edema caused by arachidonic acid and leukotriene, and meanwhile, the basil (whole herb) extract can reduce the ear swelling degree of rats and has inhibition effect on leukocyte migration in a model of air bag inflammation of rats caused by carrageenan, so that the NO and MDA content in serum is reduced and the SOD activity is enhanced; in recent years, new antibacterial mechanisms have been reported for chlorogenic acid to act by reducing the hardness of the bacterial cell wall, slowing down bacterial migration, affecting the stability of the bacterial cell membrane and inducing the production of reactive oxygen species (reactive oxygen species, ROS); research of Agarwal SK and the like shows that the active ingredients of rhein, aloe-emodin, chrysophanol and physcion in rheum officinale have antibacterial effect on novel cryptococcus, red mud fungus, candida albicans and aspergillus fumigatus. The antibacterial effective components of rhein, emodin and aloe-emodin have the most obvious effect.
When the epidermic flora of the human body is deregulated, the breeding of pathogenic bacteria can cause inflammatory reaction of the skin, and simultaneously, the generated inflammatory factors can stimulate the skin in turn, so that active oxygen free radicals are increased, larger damage is generated, and the phenomena of acne redness, epidermic water loss and the like are generated. Therefore, on one hand, the invention can inhibit the growth of harmful bacteria by regulating the biological activity of epidermic flora, reduce the risk of skin infection, and on the other hand, can reduce inflammatory reaction by reducing active oxygen free radicals, remove inflammatory factors, improve the moisture loss of horny layer and the like.
The prior researches rarely mention the application of the three components of the basil, the chlorogenic acid and the medicinal rheum officinale in the field of cosmetics.
Disclosure of Invention
The invention aims to provide a bacteriostatic and anti-inflammatory plant extract composition and application thereof, and solves the problems in the prior art. According to the invention, the antibacterial property and the additive amount of the composition are tested by taking the compound proportion of the basil, the chlorogenic acid and the medicinal rheum officinale as the cut-in points, and finally the plant extract composition with small additive amount and strong antibacterial and anti-inflammatory effects is obtained.
Aiming at the problems existing in the technical background, the invention aims to provide a preparation method of an anti-inflammatory and antibacterial plant extract composition, which comprises the following steps:
1) Pulverizing radix et rhizoma Rhei, extracting with deionized water and cellulase, and filtering to obtain first primary filtrate;
2) Extracting fresh Luo Leqie segments with ethanol to obtain a second primary filtrate;
3) Mixing the first primary filtrate and the second primary filtrate, concentrating, diluting with water, extracting with organic phase, mixing the organic phases, and concentrating to obtain fluid extract;
4) Dissolving the fluid extract with ethanol, adding chlorogenic acid powder for dissolving, concentrating to obtain concentrated solution, and ultrafiltering to obtain the anti-inflammatory and antibacterial plant extract composition;
wherein, the basil: chlorogenic acid: the mass ratio of the rhubarb raw materials is (1-3), 0.05-0.2 and (1-3).
Preferably, the preparation method comprises the following steps:
1) Pulverizing radix et rhizoma Rhei; adding deionized water for extraction, wherein the mass ratio of the rheum officinale raw material to the deionized water is 1:20-1:50, and soaking for 1-3 hours; adding cellulase at 40-50deg.C, stirring for 20 min, heating to 40-60deg.C, microwave treating for 5-15 min, reflux extracting in ultrasonic water bath for 1-3 hr, cooling to 20-30deg.C, and filtering with 100-200 mesh gauze to obtain first primary filtrate;
2) Cutting fresh basil into 1-2cm sections according to the mass ratio, soaking the fresh basil in 3-5 times of 95% ethanol solution for 1 hour, extracting at low temperature by ultrasonic oscillation, filtering, and combining the filtrates to obtain a second primary filtrate;
3) Mixing the first primary filtrate with the second primary filtrate, concentrating under reduced pressure, adding distilled water with volume 1 time of the concentrated solution, and sequentially extracting with petroleum ether, chloroform and ethyl acetate; repeating for 2-3 times until the extract is colorless, mixing ethyl acetate layers, and concentrating under reduced pressure to obtain fluid extract;
4) Dissolving the extract with 1-3 times of ethanol, adding chlorogenic acid powder according to mass ratio, stirring to dissolve completely; filtering, adding clarifier, standing, centrifuging for 30-60 min, and filtering with microfiltration membrane to obtain filtrate. Concentrating under reduced pressure to recover ethanol to obtain concentrated solution; filtering with ultrafiltration membrane to obtain the plant extract composition with antiinflammatory and antibacterial effects;
wherein the basil: chlorogenic acid: the mass ratio of the rhubarb raw materials is (1-3), 0.05-0.2 and (1-3).
Further preferably, the basil: chlorogenic acid: the mass ratio of the rhubarb raw materials is 1:0.2:2 or 1:0.1:2 or 1:0.1:3.
Further preferably, the clarifying agent in the step 4) is one or more of natural clarifying agent, chitosan, juice clarifying agent, gelatin and egg white.
Further preferably, the aperture of the microfiltration membrane in the step 4) is 0.1-1.0 μm; the aperture of the ultrafiltration membrane is 0.01-0.05 μm.
The invention also provides an anti-inflammatory and antibacterial plant extract composition, which is prepared by the preparation method;
wherein the basil: chlorogenic acid: the mass ratio of the rhubarb raw materials is 1:0.2:2 or 1:0.1:2 or 1:0.1:3.
In certain more specific embodiments, the method of preparation is specifically:
(1) Pulverizing the rheum officinale raw material according to a mass ratio;
(2) Adding deionized water for extraction, wherein the mass ratio of the raw materials to the deionized water is 1:20, and soaking for 2 hours. Adding cellulase in water bath at 48 ℃, wherein the total mass of the added cellulase accounts for 10% of the total mass of the raw materials, stirring for 20 minutes, heating to 54 ℃ for microwave treatment for 10 minutes, and carrying out ultrasonic water bath reflux extraction for 3 hours;
(3) Cooling the extracting solution obtained in the step (2) to room temperature, and filtering with 200-mesh gauze to obtain primary filtrate 1;
(4) Cutting fresh basil into 1-2cm sections according to the mass ratio, soaking the fresh basil in 3 times of 95% ethanol solution for 1 hour, extracting at low temperature by ultrasonic oscillation, filtering, and combining the filtrates to obtain primary filtrate 2;
(5) Combining the filtrates of the step (3) and the step (4), and concentrating under reduced pressure in a rotary evaporator at 55deg.C to obtain fluid extract; dissolving the extract with 1 times of distilled water, and sequentially extracting with petroleum ether, chloroform and ethyl acetate at a volume ratio of 1:1; shaking vigorously for 3h, separating organic layer, extracting for 3 times, mixing ethyl acetate layers, concentrating under reduced pressure in rotary evaporator, recovering ethyl acetate at 35deg.C to obtain fluid extract;
(6) Dissolving the extract with 3 times of ethanol, adding chlorogenic acid powder according to the mass ratio, stirring until the chlorogenic acid powder is fully dissolved, adding a clarifying agent, standing, centrifuging for 60 minutes, and filtering with a microfiltration membrane to obtain a filtrate. Filtering, concentrating under reduced pressure to recover ethanol to obtain concentrated solution.
(7) The method is characterized in that the rotating speed in the step (5) is 8000rpm;
(8) Filtering the solution obtained in the step (6) by using an ultrafiltration membrane, and collecting filtrate to obtain the plant extract composition with the effects of resisting inflammation and inhibiting bacteria.
The invention also provides application of the composition in preparation of cosmetics with anti-inflammatory and antibacterial effects.
The invention also provides a cosmetic, comprising the composition and an auxiliary agent which can be used for cosmetics.
Preferably, the composition accounts for 0.5-5% of the weight of the cosmetic.
Preferably, the cosmetic is water, emulsion, spray, cream, gel or facial mask.
The application has the following beneficial effects:
1) Can effectively inhibit the breeding of skin pathogenic bacteria, and has no influence on normal flora of skin epidermis.
2) Can reduce inflammatory reaction by reducing active oxygen free radical, scavenging inflammatory factor, improving horny layer water loss, etc.
3) Compared with the conventional antibacterial composition, the natural product is used as a raw material, so that the antibacterial composition is safer and has no irritation.
Drawings
FIG. 1 is a graph showing the effect of different compositions on IL 1-alpha secretion;
FIG. 2 is a graph showing the effect of different compositions on IL-6 secretion;
FIG. 3 is a graph showing the effect of different compositions on transdermal water loss;
FIG. 4 is a graph showing the effect of different compositions on erythema.
Detailed Description
The following will explain the embodiments of the present application in detail by way of examples, whereby the implementation process of how the present application applies technical means to solve technical problems and achieve technical effects can be fully understood and implemented accordingly.
The raw materials and equipment used in the present application are common raw materials and equipment used in the field and are commercially available products unless otherwise specified. The methods used in the present application are conventional in the art unless otherwise specified.
There are many other embodiments that can be implemented, and none of them are listed here, but the embodiments claimed in the claims of this application are all applicable.
Example 1 preparation of plant extract composition effective against inflammation and bacteriostasis
In the following examples and comparative examples of the present invention, plant extract compositions capable of effectively resisting inflammation and inhibiting bacteria were prepared by the following processes.
(1) Pulverizing the rheum officinale raw material according to a mass ratio;
(2) Adding deionized water for extraction, wherein the mass ratio of the raw materials to the deionized water is 1:20-1:50, and soaking for 1-3 hours. Adding cellulase in water bath at 40-50deg.C, wherein the total mass of the added cellulase accounts for 10% -20% of the total mass of the raw materials, stirring for 20 min, heating to 40-60deg.C, performing microwave treatment for 5-15 min, and reflux-extracting in ultrasonic water bath for 1-3 hr;
(3) Cooling the extract obtained in the step (2) to 20-30 ℃, and filtering with 100-200 mesh gauze to obtain primary filtrate 1;
(4) Cutting fresh basil into 1-2cm sections according to the mass ratio, soaking the fresh basil in 3-5 times of 95% ethanol solution for 1 hour, extracting at low temperature by ultrasonic oscillation, filtering, and combining the filtrates to obtain primary filtrate 2;
(5) Combining the filtrates of the step (3) and the step (4), and concentrating under reduced pressure in a rotary evaporator at 50-60deg.C to obtain fluid extract; dissolving the extract with 1 times of distilled water, and sequentially extracting with petroleum ether, chloroform and ethyl acetate at a volume ratio of 1:1; shaking for 2-4 hr, separating organic layer, extracting for 2-3 times until colorless, mixing ethyl acetate layers, concentrating under reduced pressure in rotary evaporator, recovering ethyl acetate at 35-50deg.C to obtain fluid extract;
(6) Dissolving the extract with 1-3 times of ethanol, adding clarifier, standing, centrifuging for 30-60 min, filtering with microfiltration membrane to obtain filtrate, and reflux extracting with 1-3 times of solution for 3-5 hr. Adding chlorogenic acid powder according to the mass ratio, and stirring until the chlorogenic acid powder is fully dissolved. Filtering, concentrating under reduced pressure to recover ethanol to obtain concentrated solution.
(7) The method is characterized in that the rotating speed in the step (5) is 5000-10000rpm;
(8) Filtering the solution obtained in the step (6) by using an ultrafiltration membrane, and collecting filtrate to obtain the plant extract composition with the effects of resisting inflammation and inhibiting bacteria.
Example 2 evaluation of Compound antibacterial Effect of different proportions of compositions
(1) Pulverizing the rheum officinale raw material according to a mass ratio;
(2) Adding deionized water for extraction, wherein the mass ratio of the raw materials to the deionized water is 1:20, and soaking for 2 hours. Adding cellulase in water bath at 48 ℃, wherein the total mass of the added cellulase accounts for 10% of the total mass of the raw materials, stirring for 20 minutes, heating to 54 ℃ for microwave treatment for 10 minutes, and carrying out ultrasonic water bath reflux extraction for 3 hours;
(3) Cooling the extracting solution obtained in the step (2) to room temperature, and filtering with 200-mesh gauze to obtain primary filtrate 1;
(4) Cutting fresh basil into 1-2cm sections according to the mass ratio, soaking the fresh basil in 3 times of 95% ethanol solution for 1 hour, extracting at low temperature by ultrasonic oscillation, filtering, and combining the filtrates to obtain primary filtrate 2;
(5) Combining the filtrates of the step (3) and the step (4), and concentrating under reduced pressure in a rotary evaporator at 55deg.C to obtain fluid extract; dissolving the extract with 1 times of distilled water, and sequentially extracting with petroleum ether, chloroform and ethyl acetate at a volume ratio of 1:1; shaking vigorously for 3h, separating organic layer, extracting for 3 times, mixing ethyl acetate layers, concentrating under reduced pressure in rotary evaporator, recovering ethyl acetate at 35deg.C to obtain fluid extract;
(6) Dissolving the extract with 3 times of ethanol, adding chlorogenic acid powder according to the mass ratio, stirring until the chlorogenic acid powder is fully dissolved, adding a clarifying agent, standing, centrifuging for 60 minutes, and filtering with a microfiltration membrane to obtain a filtrate. Filtering, concentrating under reduced pressure to recover ethanol to obtain concentrated solution.
(7) The method is characterized in that the rotating speed in the step (5) is 8000rpm;
(8) Filtering the solution obtained in the step (6) by using an ultrafiltration membrane, and collecting filtrate to obtain the plant extract composition with the effects of resisting inflammation and inhibiting bacteria.
Three plant extracts were compounded in the proportions shown in Table 1 below to give compositions 1 to 15, comparative examples 1 to 3.
TABLE 1
In the composition and the comparative example, the antibacterial and repairing effect test method and the result of the antibacterial and anti-inflammatory plant extract composition are as follows:
antibacterial effect detection
The strain is selected from: staphylococcus aureus, pseudomonas aeruginosa and propionibacterium acnes; the bacterial liquid is prepared by using pseudomonas aeruginosa, staphylococcus aureus and propionibacterium acnes as the listed strains. Preparation: diluting with sterile physiological saline to about 1.0x10 5 cfu/ml~1.0x10 6 cfu/ml of bacterial suspension. The prepared bacterial suspension should be used within 2 hours or stored at 2-8 ℃ for not more than 24 hours.
And (3) antibacterial effect detection: taking the plant composition obtained in the step (8) of example 2, taking a sterile dry filter paper sheet, and dripping 20. Mu.L of the actual plant composition into each sheet. Inoculating test bacteria: taking 1mL of each test bacterial suspension, and uniformly coating the suspension on the surface of a nutrient agar culture medium; the antibacterial test sample is pasted, a bacteria-staining flat plate is pasted for each test, the filter paper sheets are 4 sheets per dish, 3 sheets are test sample sheets, and 1 sheet is a control sample sheet and is blank. The bacteria-free tweezers are used for taking a sample, the sample is attached to the surface of a flat plate, the flat plate is placed in a constant temperature incubator at 36+/-1 ℃, the result is observed after the culture is carried out for 18 hours, and the diameter of the bacteria inhibition ring is measured by a vernier caliper. The results are the average of 3 measurements.
Experimental results: the results are shown in Table 2 below, and when compositions 1-15 are compared with comparative examples 1-3, it is found that:
1) The antibacterial activity of the 3 raw materials after being compounded and combined is superior to that of a single plant;
2) The antibacterial activity corresponding to different proportions of the 3 plant compound combinations is different, and the preferred antibacterial activity is basil: chlorogenic acid: rhubarb= (1-3): 0.05-0.2): 1-3, more preferably basil: chlorogenic acid: rhubarb = 1:0.2:2 or 1:0.1:2 or 1:0.1:3.
TABLE 2
The test results show that the compositions of test examples 1-15 have good antibacterial effects compared with the blank group, but the compositions have different proportions under the same conditions, so that the antibacterial effects on various bacteria can be influenced, and the antibacterial effects of the compositions 6, 8 and 10 are relatively good.
Example 3 detection of cellular inflammation
(1) MTT cytotoxicity test
In 96well Multiplate (burning) according to 1X10 4 The density of cells/well was inoculated with 100. Mu.L each of DMEM medium containing 10% bovine serum and keratinocytes (HaCaT), and cultured for 24 hours and then replaced with serum-free medium. Compositions 6, 8 and 10 of the above examples were added to serum-free medium, respectively, and the mixture was cultured for 24 hours after the treatment. The medium was then removed, treated with 20. Mu.L of MTT solution and allowed to react at 37℃for 2 hours. Adding 200 mu L of isopropanol into cells from which MTT solution is removed, gently shaking for 30min, completely dissolving crystalline formazan, measuring absorbance at 570nm, and calculating cell survival rate according to the following formula。
The control group was tested without the addition of sample. Cytotoxicity related results are shown in table 3 below.
TABLE 3 Table 3
(2) Inflammatory factor detection
IL 1-alpha and IL-6 are cell inflammatory factors, and detection is carried out by ELISA method. Wherein HaCat cells without any treatment were used as a blank group and Lipopolysaccharide (LPS) -induced HaCat cells were used as a model group. After stimulation of HaCat cells with 1 μg/ml LPS, 1% of compositions 6, 8, 10 were added to 12 well cell plates at 100 μl per well, at 37℃with 5% CO 2 The incubator was incubated for 72h with 3 duplicate wells per experimental group. Cell culture supernatants were collected and evaluated for anti-inflammatory efficacy by ELISA assay kit, and the results averaged. The test was as follows, taking the content detected in the blank as a reference, i.e. 100%.
The specific results are shown in FIGS. 1 and 2, FIG. 1 is the effect of different compositions on IL 1-alpha secretion; FIG. 2 is a graph showing the effect of different compositions on IL-6 secretion. The results show that the compositions 6, 8 and 10 can well inhibit the production of IL 1-alpha and IL-6 inflammatory factors after LPS stimulation, have obvious anti-inflammatory effects, and the composition 10 has the best effect.
Example 4.
In a specific embodiment, the invention provides a formula process of a mask muscle base solution capable of effectively resisting inflammation and inhibiting bacteria, wherein the specific formula is shown in the following table 4:
TABLE 4 Table 4
The extracts of claim were added to the mask base solution formulations of Table 5 below, using a formulation commonly used in the art, in an amount of 1%, to prepare mask base solution examples 1-3.
TABLE 5
Composition 6 Composition 8 Composition 10
Sample 1 Sample 2 Sample 3
1. Safety test (human body skin patch test)
15 healthy subjects with no history of dermatological allergy between 20 and 50 years of age were selected, and the patch method: selecting qualified plaque testers, dripping about 15 mu L of sample 1-3 into the plaque testers in a closed plaque test mode, sticking the sample on the back of a subject by using a special adhesive tape, sticking 20 plaque testers to each subject, respectively sticking the myobase fluid sample of the sample 1-3, removing the testers after 24 hours, observing skin reactions after removing the testers for 0.5, 6, 12, 24 and 48 hours, and recording the results according to skin reaction grade standards in skin care product health Specification.
Test results: the results of the human skin patch test show that all subjects pass the patch test and observe skin reactions at 0.5, 6, 12, 24 and 48 hours, wherein adverse reactions such as skin erythema, pimple and blister appear in 0 cases, which indicates that the product of the invention is safe and has no stimulation.
2. Skin moisture loss and erythema repair test
(1) Number of test persons: 45 people in total; divided into 5 groups of 9 persons each.
(2) Test site: the skin of the tested person with weak forearm flexor side;
(3) Sterilizing the part to be tested with 70% alcohol, and measuring skin color ERYTHEMA value (ERYHEMA) and TEWL value after alcohol is completely evaporated; selecting 1x 1cm at the skin with weaker forearm flexor side of the tested person 2 Is a control group (blank), a model group (0.075% capsaicin stimulation), and the rest is a test group. Wherein the test group was protected with the above preferred compositions 6, 8, 10 (20. Mu.L, 10 mg/mL), and was further stimulated with 0.075% capsaicin, and after 30min, skin color erythema value and TEWL value were measured to evaluate repair status.
The specific results are shown in fig. 3 and 4, and fig. 3 is the effect of different compositions on transdermal water loss; FIG. 4 is a graph showing the effect of different compositions on erythema.
Experimental results: the plant extract compositions are effective in alleviating the increase in percutaneous water loss (TEWL) and ERYTHEMA (ERYTHEMA) caused by capsaicin stimulation.
What is not described in detail in the present specification is common general knowledge of a person skilled in the art.
As used throughout the specification and claims, the word "comprise" is an open-ended term, and thus should be interpreted to mean "include, but not limited to. By "substantially" is meant that within an acceptable error range, a person skilled in the art is able to solve the technical problem within a certain error range, substantially achieving the technical effect.
It should also be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a product or system that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such product or system. Without further limitation, an element defined by the phrase "comprising one … …" does not exclude the presence of other like elements in a commodity or system comprising such elements.
While the foregoing description illustrates and describes the preferred embodiments of the present application, it is to be understood that this application is not limited to the forms disclosed herein, but is not to be construed as an exclusive use of other embodiments, and is capable of many other combinations, modifications and environments, and adaptations within the scope of the inventive concept described herein, through the foregoing teachings or through the skill or knowledge of the relevant arts. And that modifications and variations which do not depart from the spirit and scope of the present invention are intended to be within the scope of the appended claims.

Claims (11)

1. The preparation method of the anti-inflammatory and antibacterial plant extract composition is characterized by comprising the following steps of:
1) Pulverizing radix et rhizoma Rhei, extracting with deionized water and cellulase, and filtering to obtain first primary filtrate;
2) Extracting fresh Luo Leqie segments with ethanol to obtain a second primary filtrate;
3) Mixing the first primary filtrate and the second primary filtrate, concentrating, diluting with water, sequentially extracting with petroleum ether, chloroform and ethyl acetate, mixing ethyl acetate layers, and concentrating to obtain fluid extract;
4) Dissolving the fluid extract with ethanol, adding chlorogenic acid powder for dissolving, concentrating to obtain concentrated solution, and ultrafiltering to obtain the anti-inflammatory and antibacterial plant extract composition;
wherein, the basil: chlorogenic acid: the mass ratio of the rhubarb raw materials is 1:0.2:2 or 1:0.1:2 or 1:0.1:3.
2. The method of manufacturing as claimed in claim 1, comprising the steps of:
1) Pulverizing radix et rhizoma Rhei; adding deionized water for extraction, wherein the mass ratio of the rheum officinale raw material to the deionized water is 1:20-1:50, and soaking for 1-3 hours; adding cellulase at 40-50deg.C, stirring for 20 min, heating to 40-60deg.C, microwave treating for 5-15 min, reflux extracting in ultrasonic water bath for 1-3 hr, cooling to 20-30deg.C, and filtering with 100-200 mesh gauze to obtain first primary filtrate;
2) Soaking fresh Luo Leqie with 1-2cm section in 3-5 times of 95% ethanol solution for 1 hr, extracting at low temperature by ultrasonic vibration, filtering, and mixing filtrates to obtain second primary filtrate;
3) Mixing the first primary filtrate with the second primary filtrate, concentrating under reduced pressure, adding distilled water with volume 1 time of the concentrated solution, and sequentially extracting with petroleum ether, chloroform and ethyl acetate; repeating for 2-3 times until the extract is colorless, mixing ethyl acetate layers, and concentrating under reduced pressure to obtain fluid extract;
4) Dissolving the fluid extract with 1-3 times of ethanol, adding chlorogenic acid powder according to mass ratio, stirring to dissolve completely, adding clarifier, standing, centrifuging for 30-60 min, and filtering with microfiltration membrane to obtain filtrate; concentrating under reduced pressure to recover ethanol to obtain concentrated solution; filtering with ultrafiltration membrane to obtain the plant extract composition with antiinflammatory and antibacterial effects;
wherein the basil: chlorogenic acid: the mass ratio of the rhubarb raw materials is 1:0.2:2 or 1:0.1:2 or 1:0.1:3.
3. The method of claim 2, wherein the clarifying agent in step 4) is a natural clarifying agent.
4. The method of claim 2, wherein the clarifying agent in step 4) is a juice clarifying agent.
5. The method according to claim 2, wherein the clarifying agent in the step 4) is one or more of chitosan, gelatin and egg white.
6. The method according to claim 2, wherein the microfiltration membrane in step 4) has a pore size of 0.1 to 1.0 μm; the aperture of the ultrafiltration membrane is 0.01-0.05 μm.
7. An anti-inflammatory and bacteriostatic plant extract composition, characterized in that it is prepared by the preparation method according to any one of claims 1 to 6;
wherein the basil: chlorogenic acid: the mass ratio of the rhubarb raw materials is 1:0.2:2 or 1:0.1:2 or 1:0.1:3.
8. Use of a composition according to claim 7 for the preparation of a cosmetic product having anti-inflammatory and bacteriostatic effects.
9. A cosmetic comprising the composition of claim 7 and a cosmetically acceptable adjuvant.
10. The cosmetic according to claim 9, wherein the composition comprises 0.5 to 5% by mass of the cosmetic.
11. The cosmetic product of claim 9, wherein the cosmetic product is an aqueous solution, an emulsion, a spray, a cream, a gel, or a mask.
CN202110771103.8A 2021-07-07 2021-07-07 Preparation and application of antibacterial and anti-inflammatory plant extract composition Active CN113712858B (en)

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