CN111973515B - Bacteriostatic restoration plant extraction composition, preparation method and application thereof - Google Patents
Bacteriostatic restoration plant extraction composition, preparation method and application thereof Download PDFInfo
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Abstract
The invention provides an antibacterial restoration plant extraction composition, a preparation method and application thereof. The plant extract composition comprises the following components in percentage by mass: fructus forsythiae: houttuynia cordata: isatis root (1-6.5): (1-6.5): 1-6.5). The plant extract composition is prepared by the processes of crushing, extracting, concentrating, formulating, sterilizing and the like, and can obviously inhibit common pathogenic bacteria. The raw materials of the invention are pure natural plants, the synergistic effect of each component is good, the invention has obvious bacteriostasis and repair effects, and the invention has mild property, easy absorption and no stimulation, particularly meets the requirements of people on safety, effectiveness and zero burden of cosmetics, and can be widely applied to skin care cosmetics.
Description
Technical Field
The invention belongs to the field of cosmetics, and particularly relates to a plant extract composition with antibacterial and repairing effects, and a preparation method and application thereof.
Background
The demand of the current market for bacteriostatic products is greatly increased, and bacteriostatic and disinfectant products become indispensable articles for daily use for people in this era. On the other hand, with the improvement of living standard and the enhancement of health consciousness, people pay more attention to the green safety and functionality of medicines and cosmetics, natural and healthy plant extract products are popular and advocated by more and more consumers, and the market is gradually expanded. Therefore, among today's various types of bacteriostatic products, bacteriostatic products derived from plant extracts are popular among consumers due to their advantages of greenness and safety. At the same time, however, plant extracts selected from natural plants and applied to the field of cosmetic applications often face problems of too high addition levels or too complicated extraction and purification processes.
Fructus forsythiae (Forsythia suspensa) is a common traditional Chinese medicine for clearing heat and removing toxicity, is a shrub of filial leaf of Forsythia of dicotyledonae, twisted flower order, Oleaceae and Forsythia, contains active ingredients such as forsythol, forsythin lipoid glycoside, oleanolic acid, vitamin P and the like, and has the functions of bacteriostasis, anti-inflammation, anti-allergy and the like through modern pharmacological research; houttuynia cordata, also called herba houttuyniae root, is a dry aerial part of a perennial herb Houttuynia cordata (Houttuynia cordiata Thunb.) of Saururaceae, has the effects of clearing away heat and toxic materials, relieving swelling and treating sore, and is found to have the effects of bacteriostasis, antivirus, improving the immunity of the organism, diuresis and the like by modern pharmacological research; radix Isatidis is Isatis Indigotica fortunei (Isatis indigotica fortunei) of Brassicaceae and has effects of clearing away heat and toxic materials, cooling blood, removing macula, relieving sore throat and relieving pain. Modern pharmacology finds that the traditional Chinese medicine composition has the effects of bacteriostasis, antivirus, detoxification and the like.
At present, the compound application of the three plants mainly focuses on the aspects of Chinese medicinal preparations and oral administration. For example, CN 102429995a discloses a preparation method of a compound houttuynia cordata mixture, which is an application of a stable Chinese medicinal preparation obtained by a decoction alcohol precipitation process, and solves the problem of precipitation generated in the long-term standing process of the existing compound houttuynia cordata mixture. CN 105168509A discloses a preparation process of a high-content compound houttuynia cordata mixture, which can effectively improve the content of drug-effect components such as houttuynine sodium bisulfite, quercetin and the like, remove components such as polysaccharide, protein, tanning and the like in medicinal materials, and improve the stability and safety of the medicine.
Extracts from forsythia fruit, isatis root or houttuynia cordata are reported in the bacteriostasis aspect, but have the problems of unsatisfactory bacteriostasis effect, overhigh effect concentration, risk of organic solvent residue and the like. Gaojie and the like (Gaojie and the like, experimental research on the in-vitro bacteriostatic action of 10 Chinese herbal medicines on clinically common pathogenic bacteria [ J ]. famous physicians, 2019(10):242) finds that fructus forsythiae has no obvious bacteriostatic effect or no bacteriostatic effect on Escherichia coli and staphylococcus aureus, and the diameters of bacteriostatic rings are respectively 8mm and 5mm (wherein, the diameter of the bacteriostatic ring is less than 6mm, the bacteriostatic activity is not existed, and the diameter of the bacteriostatic ring is more than 6mm, the bacteriostatic activity is existed).
Patent document CN 111035668A discloses the use of an instant isatis leaf extract in the treatment of psoriasis and psoriasis, and discloses that the isatis leaf extract powder (sample concentration 60mg/mL) has a significant bacteriostatic effect on staphylococcus aureus, escherichia coli and candida albicans (zone diameters of 11mm, 10mm and 13mm, respectively). The research of the aged gem and the like (the aged gem and the like, the analysis of main components of forsythia suspense seeds in different production places and the bacteriostasis research [ J ]. Shizhen national medicine, 2018,29(02):427 one 430) finds that the forsythia suspense seeds have different degrees of inhibition on escherichia coli, staphylococcus aureus and bacillus subtilis, but the sample concentration is as high as 200 mg/mL. If the product is applied to the field of cosmetics, problems of high cost, stability and the like exist, so that the application thereof is limited.
Zhengdong excess (Zhengdong excess, study on bacteriostatic effect of houttuynia cordata on staphylococcus aureus and escherichia coli [ J ] experimental science and technology, 2019,17(04): 103-. Wherein, the direct grinding method and the water extraction method have no obvious inhibition effect on the escherichia coli, and the organic solvent extraction method has obvious inhibition effect on the escherichia coli. However, when the organic solvent extraction process is applied to large-scale production, the risk of organic solvent residue exists, and the safety of the product is further influenced, so that the application range of the method is limited.
The application of the composition of fructus forsythiae, radix isatidis and herba houttuyniae in the antibacterial activity in the field of cosmetics is rarely mentioned in the existing research, so the research of the antibacterial activity and the addition amount of the composition finally obtained by compounding the fructus forsythiae, the radix isatidis and the herba houttuyniae is an entry point to carry out relevant tests. Therefore, the need in the art is to provide a plant extract composition with low dosage and strong bacteriostatic and repairing effects, and a preparation method, application and additive advantages thereof.
Disclosure of Invention
The invention aims to provide the bacteriostatic plant extract composition with low addition and high effect under a certain processing technology by taking the application of fructus forsythiae, herba houttuyniae and radix isatidis in the bacteriostatic property of the cosmetic field as a target, taking the compounding ratio of the fructus forsythiae, the herba houttuyniae and the radix isatidis as an entry point, and taking the bacteriostatic property and the addition amount of the composition.
The invention provides a bacteriostatic and repairing plant extraction composition, which consists of the following components in percentage by mass: fructus forsythiae: houttuynia cordata: isatis root (1-6.5): (1-6.5): 1-6.5).
The forsythia suspense fruit: houttuynia cordata: the mass percentage of the isatis root is (1-3) to (1-3).
The forsythia suspense fruit: houttuynia cordata: the mass percentage of the isatis root is 1:3: 2.
The forsythia suspense fruit: houttuynia cordata: the mass percentage of the isatis root is 1:2: 1.
The forsythia suspense fruit: houttuynia cordata: the mass percentage of the isatis root is 1:2: 2.
In a second aspect, the invention provides a bacteriostatic repair composition, which contains 0.2-5 wt% of the plant extract composition and an excipient. The excipient may be a liquid excipient or a solid excipient. Such as binders, fillers, disintegrants, lubricants in tablets; wine, vinegar, medicinal juice, etc. in the Chinese medicinal pill; base portion in semisolid formulations ointments, creams; liquid preparations such as preservatives, antioxidants, flavors, fragrances, cosolvents, emulsifiers, solubilizers, tonicity modifiers, colorants and the like in cosmetics can be referred to as excipients or as adjuvants. The weight percentage of the plant extract composition is 0.5-3%.
The bacteriostatic repairing composition has inhibitory effect on Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Propionibacterium acnes. More preferably, the bacteriostatic effect on staphylococcus aureus, escherichia coli and pseudomonas aeruginosa is better.
In a third aspect, the present invention provides the use of the plant extract composition described above in the preparation of a cosmetic.
The cosmetic is in the form of aqua, emulsion, spray, cream, gel or facial mask.
In a fourth aspect, the present invention provides a method for preparing the above plant extract composition, the method comprising the steps of:
(1) respectively crushing the plant raw material medicines, weighing according to the proportion, and uniformly mixing;
(2) adding deionized water for extraction at a mass ratio of 1:20-1:50, at 75-95 deg.C for 1-3h for 1-3 times;
(3) cooling the extracting solution obtained in the step (2) to 20-30 ℃, and filtering with 100-200-mesh gauze to obtain primary filtrate;
(4) adding a clarifying agent into the primary filtrate obtained in the step (3), and standing at a low temperature for 6-8 h;
(5) centrifuging the solution obtained in the step (4) for 20-60min at the rotation speed of 5000-;
(6) concentrating the secondary filtrate obtained in the step (5) under reduced pressure, adding polyol, and stirring or ultrasonically mixing uniformly;
(7) and (4) filtering the solution obtained in the step (6) by using an ultrafiltration membrane, and collecting filtrate to obtain the bacteriostatic and repairing plant extract composition.
Preferably, the clarifying agent in the step (4) is one or more of a ZTC101 natural clarifying agent, chitosan, a 101 fruit juice clarifying agent, gelatin and egg white;
the aperture of the microfiltration membrane in the step (5) is 0.1-1.0 μm;
concentrating the secondary filtrate obtained in the step (6) to 1/2-1/5 of the original weight, wherein the addition amount of the polyhydric alcohol is 1-10 times of that of the concentrated solution, and the polyhydric alcohol is one or more of 1, 3-butanediol, 1, 2-propylene glycol, dipropylene glycol and glycerol;
the aperture of the ultrafiltration membrane in the step (7) is 0.01-0.05 μm.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the invention adopts gauze-microfiltration-ultrafiltration step-by-step filtration, realizes the purposes of separation, purification and refining in a combined way, and has the advantages of simple and convenient operation, low energy consumption, short extraction time, better effect and more stable extracting solution compared with the traditional method.
(2) The invention provides a plant extraction composition with antibacterial and repairing effects, which is prepared from natural plants, utilizes the traditional medical characteristics of fructus forsythiae, herba houttuyniae and radix isatidis, and combines the existing new technology to realize a new idea in the field of cosmetics antibacterial. The three plants have synergistic effect after being combined, can achieve the advantages of good bacteriostatic effect, multiple bacteriostatic types and small addition amount, is a safe, non-irritant and easily-absorbable natural plant composition, and meets the requirements of people on active ingredients and effects of cosmetics.
Drawings
FIG. 1 is a graph of the bacteriostatic and reparative effects of compositions 8, 14, 15 and comparative examples 1-3;
FIG. 2 is a test chart of a patch on human skin (composition 8);
FIG. 3 relief effects of capsaicin-induced ERYTHEMA for compositions 8, 14, 15 and comparative examples 1-3.
FIG. 4 the alleviating effect of compositions 8, 14, 15 and comparative examples 1-3 on capsaicin-induced TEWL.
Detailed Description
Hereinafter, the present invention is described in more detail and specifically with reference to examples, but the following examples are not intended to limit the present invention.
In the present invention, all the equipment and materials are commercially available or commonly used in the art, and the methods in the following examples are conventional in the art unless otherwise specified.
Example 1 preparation of bacteriostatic plant extract composition
In the following examples and comparative examples of the present invention, bacteriostatic plant extract compositions were prepared using the following processes.
(1) Pulverizing fructus forsythiae, herba Houttuyniae, and radix Isatidis respectively, and mixing;
(2) adding deionized water for extraction, wherein the mass ratio of the raw material medicine to the deionized water is 1:30, the extraction temperature is 90 ℃, the extraction time is 1.5h, and the extraction times are 2 times;
(3) cooling the extracting solution obtained in the step (2) to 25 ℃, and filtering the extracting solution by using 200-mesh gauze to obtain primary filtrate;
(4) adding clarifying agent chitosan into the primary filtrate obtained in the step (3), and standing for 8 hours at low temperature;
(5) centrifuging the solution obtained in the step (4) for 40min at the rotating speed of 5000rpm, filtering with a microfiltration membrane with the pore diameter of 0.2 mu m, and collecting filtrate to obtain secondary filtrate;
(6) concentrating the secondary filtrate obtained in the step (5) under reduced pressure to 1/2 of the original weight, adding polyol 2 times the weight of the concentrated solution, and stirring or ultrasonically mixing uniformly;
(7) and (4) filtering the solution obtained in the step (6) by using an ultrafiltration membrane with the pore diameter of 0.05 mu m, and collecting filtrate, namely the plant extract composition with the antibacterial and repairing effects.
The addition amount of the polyhydric alcohol in the step (6) is 1-10 times of the concentrated solution, and the polyhydric alcohol is one or more of 1, 3-butanediol, 1, 2-propanediol, dipropylene glycol and glycerol. Compositions 1-15 and comparative examples 1-3, the above-mentioned method was used to prepare each of the extract compositions 1-15 according to the proportions in table 1, and the compositions obtained at the proportions were tested for bacteriostatic and reparative effects and safety according to the bacteriostatic test methods described below.
TABLE 1 compositions and comparative example formulations
Effect verification
In the composition and the comparative example of the invention, the method for testing the bacteriostasis and repair effect of the bacteriostatic plant extract composition and the results are shown in the subsequent examples.
Example 2 detection of bacteriostatic Effect
Selecting strains: staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, propionibacterium acnes; the preparation of the bacterial liquid selects pseudomonas aeruginosa, staphylococcus aureus, escherichia coli and propionibacterium acnes as the listed strains. Preparation: by usingDiluting with physiological saline to about 1.0x10 5 cfu/ml~1.0x10 6 cfu/ml bacterial suspension. The prepared bacterial suspension can be used within 2h, or can be stored at 2-8 ℃ for no more than 24 h.
And (3) detecting the antibacterial effect: the effective plant composition obtained in the step (7) of example 1 was taken, and sterile dried filter paper sheets were each dropped with 20. mu.L of a plant composition solution (10mg/mL) at an actually used concentration. Test bacterium inoculation: taking 1mL of each test bacterial suspension, and uniformly smearing the test bacterial suspension on the surface of a nutrient agar culture medium; the bacteriostatic test samples are attached, one bacterial staining flat plate is attached to each test, 4 filter paper sheets are attached to each dish, 3 filter paper sheets are test sample sheets, and 1 filter paper sheet is blank. A sample piece is attached to the surface of the flat plate by using a sterile forceps, the flat plate is placed in a constant temperature incubator at 36 +/-1 ℃, the result is observed after the flat plate is cultured for 18 hours, and the diameter of the antibacterial ring is measured by using a vernier caliper. The results are the average of 3 measurements.
The experimental results are as follows: the results are shown in Table 2 and FIG. 1, and when compositions 1-15 are compared with comparative examples 1-3, it is found that:
1) the bacteriostatic activity of the plant compound combination is superior to that of a single plant;
2) the antibacterial activities corresponding to different proportions of the plant compound combination are different, and the plant compound combination is preferably forsythia fruit: houttuynia cordata: isatis root (1-3): (1-3): 1-3), more preferably forsythia fruit: houttuynia cordata: isatis root is 1:3:2 or 1:2:1 or 1:2: 2.
TABLE 2 compositions 1-15 and comparative examples 1-3 bacteriostatic effect
Example 3 cytotoxicity assessment assay:
MTT assay is an experimental method widely used for detecting cytotoxicity or cell proliferation by measuring the number of living cells, and the detection principle is succinate dehydrogenase (succinate dehydrogenase) in mitochondria of living cellsrogenase or mitochondril dehydrogenase) makes the water-soluble yellow salt MTT (3- [4, 5-dimethylthiozole-2-yl)]2,5-diphenyl tetrazolium bromide) to a water-insoluble blue color and deposited in the cells, whereas dead cells do not have this function. The resulting crystalline formazan (formazan) is generally dissolved in dmso (dimethlysulfoxide) and then the absorbance is measured. The specific experimental method is as follows 1X10 in 96well multi plate (corning) 4 cells/well were seeded at a density of 100. mu.L each in DMEM medium containing 10% bovine serum and keratinocytes (HaCaT), and 24 hours later in culture were replaced with serum-free medium. Samples with different concentrations were added to serum-free medium for 24 hours after treatment. Thereafter, the medium was removed, treated with 20. mu.L of MTT solution, and allowed to react at 37 ℃ for 2 hours. The MTT solution was removed and 200. mu.L of isopropanol was added to the cells and shaken gently for 30min to dissolve the crystalline formazan completely, absorbance was measured at 570nm, and the cell viability was calculated according to the following formula.
The control group was tested without the addition of sample. The cytotoxicity results are shown in table 3.
TABLE 3 MTT test results for compositions 1-15 and comparative examples 1-3
Example 4 safety Patch test
Dripping 20 mu L of liquid to be detected into the spot tester, wherein a blank control hole is used as a blank control (pure water); the spot tester with the tested object is attached to the bent side of the forearm of the tested person, and the tested person is lightly pressed by the palm to be uniformly attached to the skin for 24 hours; the skin irritation and sensitization were observed as shown in table 2 after 30min, 24 hours, and 48 hours, respectively, after removal of the test substance patch tester, and the observation results were recorded.
TABLE 4 skin reaction grading Standard for skin Enclosed Patch test
The experimental results are as follows: the results of the human skin patch tests performed on compositions 8, 14, 15 are shown in table 5 and figure 2. The result of the skin patch test of the human body shows that 20 people have no adverse reaction to the skin.
TABLE 5 cosmetic human skin Patch test results
Example 5 skin Barrier repair test
(1) The number of tested persons: 20 persons;
(2) the tested part: the skin of the forearm flexor side of the tested person;
(3) sterilizing the part to be tested with 70% ethanol, and measuring skin color ERYTHEMA value (ERYTHEMA) and TEWL value after the ethanol is completely evaporated; selecting 1x 1cm on weak skin of forearm of human subject 2 The areas of (a) are control (blank), model (0.075% capsaicin challenge), and the rest are test groups. Wherein the test groups were protected with the preferred compositions 8, 14, 15 or comparative examples 1-3(20 μ L, 10mg/mL) as described above, and then were challenged with 0.075% capsaicin, and skin color erythema values and TEWL values were measured after 30min to evaluate the repair status.
The experimental results are as follows: the results are shown in fig. 3-4, and the respective plant extract compositions are effective in relieving the rise of percutaneous water loss (TEWL) and the increase of ERYTHEMA (ERYTHEMA) due to capsaicin stimulation, whereas the comparative examples have limited relieving effects on TEWL and ERYTHEMA.
Further, it should be understood that the above embodiments are only intended to illustrate the technical solutions of the present invention, and not to limit the scope of the present invention, and although the present invention has been described in detail with reference to the preferred embodiments, it should be understood by those skilled in the art that various modifications or equivalent substitutions may be made thereto without departing from the scope and spirit of the present invention.
Claims (11)
1. The bacteriostatic and repairing plant extraction composition is characterized by comprising the following components in percentage by mass: fructus forsythiae: houttuynia cordata: 1-6.5 of isatis root, 1-6.5 of isatis root and 1-6.5 of isatis root; the preparation method of the extraction composition comprises the following steps:
(1) respectively crushing the plant raw material medicines, weighing according to the proportion, and uniformly mixing;
(2) adding deionized water for extraction at a mass ratio of 1:20-1:50, at 75-95 deg.C for 1-3h for 1-3 times;
(3) cooling the extracting solution obtained in the step (2) to 20-30 ℃, and filtering with 100-200-mesh gauze to obtain primary filtrate;
(4) adding a clarifying agent into the primary filtrate obtained in the step (3), and standing at a low temperature for 6-8 h;
(5) centrifuging the solution obtained in the step (4) for 20-60min at the rotation speed of 5000-;
(6) concentrating the secondary filtrate obtained in the step (5) under reduced pressure, adding polyol, and stirring or ultrasonically mixing uniformly;
(7) and (4) filtering the solution obtained in the step (6) by using an ultrafiltration membrane, and collecting filtrate to obtain the bacteriostatic and repairing plant extract composition.
2. The plant extract composition of claim 1, wherein the ratio of forsythia suspense seed: houttuynia cordata: the mass percentage of the isatis root is (1-3) to (1-3).
3. The plant extract composition of claim 1, wherein the ratio of forsythia suspense seed: heartleaf houttuynia herb: the mass percentage of the isatis root is 1:3: 2.
4. The plant extract composition of claim 1, wherein the ratio of forsythia suspense seed: houttuynia cordata: the mass percentage of the isatis root is 1:2: 1.
5. The plant extract composition of claim 1, wherein the ratio of forsythia suspense seed: houttuynia cordata: the mass percentage of the isatis root is 1:2: 2.
6. The plant extract composition as claimed in any one of claims 1 to 5, wherein the clarifying agent in step (4) is one or more of ZTC101 natural clarifying agent, chitosan, 101 juice clarifying agent, gelatin, and egg white;
the aperture of the microfiltration membrane in the step (5) is 0.1-1.0 μm;
concentrating the secondary filtrate obtained in the step (6) to 1/2-1/5 of the original weight, wherein the addition amount of the polyhydric alcohol is 1-10 times of that of the concentrated solution, and the polyhydric alcohol is one or more of 1, 3-butanediol, 1, 2-propylene glycol, dipropylene glycol and glycerol;
in the step (7), the aperture of the ultrafiltration membrane is 0.01-0.05 μm.
7. A bacteriostatic repairing composition, which comprises 0.2-5 wt% of the plant extract composition of any one of claims 1-5 and an excipient.
8. The bacteriostatic repairing composition according to claim 7, wherein the weight percentage of the plant extract composition is 0.5-3%.
9. The bacteriostatic restoration composition according to claim 7, wherein the bacteriostatic restoration composition has inhibitory effect on Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis and Propionibacterium acnes.
10. Use of the plant extract composition according to any one of claims 1 to 5 for the preparation of a cosmetic product.
11. Use of a plant extract composition according to claim 10, wherein the cosmetic is a water, an emulsion, a spray, a cream, a gel or a mask.
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CN103494928A (en) * | 2013-09-29 | 2014-01-08 | 江西天施康中药股份有限公司 | Preparation method for compound houttuynia cordata preparation |
CN106063793A (en) * | 2015-04-23 | 2016-11-02 | 富力 | The application in preparation treatment antibacterial infection medicine of phillyrin, its derivant, the phillyrin/phillygenol compositions |
WO2018112941A1 (en) * | 2016-12-23 | 2018-06-28 | 王晓伟 | Multi-component composite nonwoven fabric |
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