CN117122657A - Chinese medicinal anti-dandruff composition, and preparation method and application thereof - Google Patents
Chinese medicinal anti-dandruff composition, and preparation method and application thereof Download PDFInfo
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- CN117122657A CN117122657A CN202311306971.4A CN202311306971A CN117122657A CN 117122657 A CN117122657 A CN 117122657A CN 202311306971 A CN202311306971 A CN 202311306971A CN 117122657 A CN117122657 A CN 117122657A
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Abstract
The invention relates to a traditional Chinese medicine anti-dandruff composition, which comprises the following components in parts by weight: 1.0 to 3.0 percent of giant knotweed extract, 1.5 to 2.5 percent of Baistep extract, 1.5 to 4.0 percent of pseudo-ginseng extract, 2.0 to 3.0 percent of rice ferment and the balance of process water. The composition has the effects of inhibiting staphylococcus epidermidis, reducing scalp inflammation and removing dandruff.
Description
Technical Field
The invention belongs to the technical field of traditional Chinese medicine anti-dandruff, and in particular relates to a traditional Chinese medicine anti-dandruff composition, and a preparation method and application thereof.
Background
Dandruff is produced by the continuous exfoliation of the horny layer of the scalp, a result of metabolism. However, when the dandruff is too much, the type of dandruff is changed from physiological dandruff to pathological. Studies have shown that 60% of Chinese adults suffer from dandruff-related problems, with 15% of dandruff-related symptoms being severe.
Currently, in the daily chemical market, the anti-dandruff product accounts for 35%. But 70% of the population is not satisfied with the anti-dandruff effect.
Simple treatments for dandruff are mainly performed using shampoos containing antifungal active ingredients (i.e. antidandruff agents), the use of which can act to effectively inhibit and reduce dandruff. Currently, the anti-dandruff agents developed in the market mainly comprise pyrithione (ZPT), ganbazole, pyridone ethanolamine salt (OCT), ketoconazole and the like, but the chemical reagent products have high toxicity and irritation, are easy to generate drug resistance after long-term use, and can bring unpredictable damage to the immune system of a human body.
In the aspect of scalp health management, the traditional Chinese medicine has natural advantages, and researches report that certain Chinese herbal medicines can effectively treat skin inflammation to play a role of removing dandruff, and in treating head seborrheic dermatitis, a good clinical effect is obtained, but the traditional Chinese medicine has the problem of only single anti-dandruff effect and is not ideal.
In general, although the incidence of dandruff is high, the mechanism of its occurrence is not yet clear. Recent views suggest that there is an important correlation between dandruff and scalp flora micro-ecology and stratum corneum barrier; not just the result of malassezia alone (the population of dandruff after use of shampoo products having anti-malassezia activity does not completely remove the dandruff, which suggests that the production of dandruff may involve a plurality of microorganisms, rather than malassezia alone. Therefore, other scalp microorganisms and new directions of antagonizing inflammatory reaction to protect scalp cuticle barrier are important to prepare efficient anti-dandruff products.
In view of the analysis, the scalp can be healthily nourished only by comprehensively, multi-angularly and scientifically and orderly conditioning and curing the scalp, so that the purpose of removing dandruff from the root is achieved. In the process, the combination compatibility of the traditional Chinese medicines has wide application prospect.
Disclosure of Invention
In order to overcome the problems in the background art, the invention provides a traditional Chinese medicine anti-dandruff composition, and a preparation method and application thereof.
In order to achieve the above purpose, the invention is realized by the following technical scheme:
the invention provides a traditional Chinese medicine anti-dandruff composition, which comprises the following components in parts by weight: 1.0 to 3.0 percent of giant knotweed extract, 1.5 to 2.5 percent of Baistep extract, 1.5 to 4.0 percent of pseudo-ginseng extract, 2.0 to 3.0 percent of rice ferment and the balance of process water.
In a second aspect, the present invention provides a process for the preparation of a composition as described above, comprising the steps of:
s1, preparation of an extract: the traditional Chinese medicine materials are prepared by the following steps: respectively placing rhizoma Polygoni Cuspidati, radix et rhizoma Rhei, notoginseng radix and rice yeast in percolating tank, adding 70% ethanol 5 times of the weight of the materials, soaking at room temperature for 48 hr, and percolating to obtain extractive solution; adding 60% ethanol with 5 times of the weight of the medicinal materials into a percolation tank respectively, soaking for 48 hours, percolating to obtain extract, mixing the two extracts, and uniformly mixing to obtain extract of different Chinese medicinal materials;
s2, concentrating an extracting solution: at the temperature: 72.+ -. 1 ℃ and vacuum degree: -0.05-0.06 mpa, rotational speed: 30+/-1 rpm, adding the uniformly mixed extracting solution into concentrating equipment, and concentrating to 30% of the initial mass to obtain concentrated solutions of all components;
s3, blending: stirring and heating the concentrated solutions of all the components to 80-85 ℃ at the rotating speed of 40+/-5 rpm, adding the mixed solution into a wide-mouth container according to a proportion, mixing, heating in a water bath, keeping the temperature at 80-85 ℃, preserving heat for 1h in an open way, and continuously stirring during heating and preserving heat to concentrate the concentrated solution to 50% of the mass during adding;
s4, stirring and mixing uniformly after heat preservation is finished, and quantifying the process water to 100% to obtain the traditional Chinese medicine anti-dandruff composition.
In a third aspect, the present invention provides the use of the above composition in an external preparation for the prevention and treatment of dandruff or in a scalp care product.
In a fourth aspect the invention provides the use of a composition as described above in a product for inhibiting staphylococcus epidermidis and reducing scalp inflammation.
The invention has the beneficial effects that: the invention has the effects of inhibiting staphylococcus epidermidis, reducing scalp inflammation and removing dandruff.
Drawings
FIG. 1 is a comparison of scalp condition before and after use of the antidandruff composition according to the invention, in example 5 of the present invention, specific case 1.
Figure 2 is a comparison of scalp condition before and after use of the antidandruff composition according to the invention in example 5 of the present invention.
FIG. 3 is a comparison of scalp condition before and after use of the antidandruff composition according to the invention, according to example 5 of the present invention, according to example 3.
Detailed Description
In order to make the objects, technical solutions and advantageous effects of the present invention more apparent, preferred embodiments of the present invention will be described in detail below to facilitate understanding by the skilled person.
Example one, preparation of Chinese herbal extracts and compositions:
s1, preparation of an extract: the traditional Chinese medicine materials are prepared by the following steps: respectively placing rhizoma Polygoni Cuspidati, radix et rhizoma Rhei, notoginseng radix and rice yeast in percolating tank, adding 70% ethanol 5 times of the weight of the materials, soaking at room temperature for 48 hr, and percolating to obtain extractive solution; adding 60% ethanol with 5 times of the weight of the medicinal materials into a percolation tank respectively, soaking for 48 hours, percolating to obtain extract, mixing the two extracts, and uniformly mixing to obtain extract of different Chinese medicinal materials;
s2, concentrating an extracting solution: at the temperature: 72.+ -. 1 ℃ and vacuum degree: -0.05-0.06 mpa, rotational speed: 30+/-1 rpm, adding the uniformly mixed extract into a concentrating device, concentrating to 30% of the initial mass, and obtaining concentrated solutions of all components to obtain the traditional Chinese medicine extract;
s3, blending: stirring and heating the concentrated solutions of all the components to 80-85 ℃ at the rotating speed of 40+/-5 rpm, adding the components into a wide-mouth container according to the corresponding weight fraction, mixing, heating in a water bath, keeping the temperature at 80-85 ℃, preserving heat for 1h in an open way, and continuously stirring in the heating and heat preserving processes to concentrate the concentrated solution to 50% of the mass during the adding;
s4, stirring and mixing uniformly after heat preservation is finished, and quantifying the process water to 100% to obtain the traditional Chinese medicine anti-dandruff composition.
Example two, screening of the Chinese medicinal composition of the invention for antagonizing Staphylococcus epidermidis Activity
1. Experimental materials and reagents
1.1 samples were prepared with physiological saline.
1.2 test strains: staphylococcus epidermidis ATCC 12228.
1.3 Medium: TSB solid medium: peptone 10g/L, naCl 10g/L, K 2 HPO 4 2.5 g/L, yeast extract 3g/L, agar 15g/L, distilled water to dissolve, regulating pH to 7.2+ -0.2, and autoclaving at 115 deg.C for 20min; TSB broth: peptone 10g/L, naCl 10g/L, K 2 HPO 4 2.5 g/L, yeast extract 3g/L, distilled water to dissolve, adjusting pH to 7.2+ -0.2, and autoclaving at 115 deg.C for 20min.
1.4 consumable: culture dish, erlenmeyer flask, oxford cup, pipette, 96 well culture plate, polypropylene tube, test tube, and coating rod.
Method
2.1 preparation of bacterial suspension and inoculation:
coating 100 μl of the staphylococcus epidermidis preserved in glycerol on nutrient agar solid medium, culturing at 35deg.C for 1d, re-suspending with physiological saline to obtain bacterial suspension, counting by plate culture method, and adjusting test bacterial concentration to 10 5 ～10 6 CFU/mL。
2.2 determination of the antibacterial action of the samples
The culture dish was poured with TSB agar solid medium, and after solidification, 100. Mu.L of test bacteria was uniformly spread on the medium. 2 sterile oxford cups were placed equidistantly on the medium: the inner diameter (6.0+/-01) mm, the outer diameter (8.0+/-0.1) mm and the height (10.0+/-0.1) mm, 200 mu L of samples with different concentrations (0.5-6%) to be tested are respectively injected into the cup, and physiological saline is used as a negative control. After culturing in an incubator at 35 ℃ for 2d, the diameter of the inhibition zone of each medicine was measured by a ruler, and the average value was obtained.
2.3 determination of minimum inhibitory concentration (MIC value)
Preparing a liquid medicine:
and diluting the sample to be tested into experimental concentration by using physiological saline.
Preparation of TSB liquid culture medium dilution bacterium from bacterium liquidSuspension to a final bacterial concentration of l0 5 CFU/mL。
Culturing and judging the results, wherein 100 mu L of bacterial liquid and 100 mu L of sample (the mass percentage of active ingredients in the sample is 0.5%, 1%, 2%, 4%, 6%, 8%, 10% and 12%) are added into the micro-holes, meanwhile, a negative control without bacteria addition and a normal growth control without liquid medicine addition are arranged, and 3 parallel medicines are used for each medicine, and the average value is obtained. After incubation in a wet box at 30 ℃ for 48 hours, the results are observed and the data are read by a direct method. The result is judged that the premise is that the growth control is good, the blank control is sterile and clear in growth, and other holes are inhibited along with the increase of the concentration gradient of the medicine.
Results
The results are shown in table 1 below:
as can be seen from the results in Table 1, the extracts of Polygonum cuspidatum and Stemonae radix have relatively obvious activities of antagonizing Staphylococcus epidermidis; whereas the notoginseng extract and the rice ferment extract show substantially no antagonistic activity against staphylococcus epidermidis.
Embodiment III A traditional Chinese medicine formula unilateral antagonizing inflammation experiment
1. Method of
The experimental method comprises the following steps: human epidermal keratinocytes were stimulated with 0.01% sebum for 24h, then RNA was collected using Shanghai-associated bioengineering Co., ltd. Kit Ultra Pure Total RNA Fast Extraction Kit, cDNA was synthesized using reverse transcription kit nvitrogen SuperScript Kits, and finally real-time fluorescent quantitative PCR was performed using FastStart Essential DNAGreen Master to test the gene expression amount of inflammatory factor mRNA.
The mRNA expression level of the target gene is calculated by a 2-delta Ct method by taking the internal reference gene actin as a standard and taking delta Ct which is the lowest value of the expression level of the target gene in all experimental data as a contrast.
Results
The results are shown in Table 2 below:
as can be seen from table 2, the extracts of polygonum cuspidatum, stemona root and notoginseng all showed different degrees of activity compared to the positive control oleic acid; wherein, the notoginseng extract has obvious inhibition effect on the mRNA gene expression of the keratinocyte inflammatory factor.
Fourth embodiment, orthogonal combination experiment of the invention
Experimental materials and reagents: same as in example 1
2. The method comprises the following steps:
2.1 composition combinatorial optimization: design using a 5-factor 4 horizontal orthogonal table (with a set of more than one as the error bars)
2.2 determination of the antibacterial Rate of the composition
Washing the test bacteria 24h slant culture with PBS to obtain bacterial suspension (required concentration is that 100 μl is dropped into control 5mL sample solution, and the number of recovered bacteria is 5×10) 4 CFU/mL). The sample solution (5 mL) and the control sample were taken and passed through a 4-tube. Taking the bacterial suspension, respectively dripping 100 mu L of the bacterial suspension into each tested combined sample solution and each control sample uniformly, putting into a test tube containing 5mL of PBS, fully mixing uniformly, properly diluting, and taking 10 parts of the bacterial suspension -4 ～10 -5 The dilution is carried out by respectively sucking 0.5mL, placing in 2 plates, pouring 15mL of nutrient agar medium (bacteria) cooled to 40-45 ℃ into the plates, rotating the plates to make the plates fully uniform, overturning the plates after agar solidification, and culturing for 48h at 30+/-2 ℃. Viable bacterial colony counts were made. The test was repeated 2 times.
Calculating the bacteriostasis rate: x= (Nc-Ns)/nc×100%. Wherein: x is the antibacterial rate; NC is the average colony number of the control sample, CFU/dish; NS is the average colony count of the test sample, CFU/dish.
2.3 Measurement of the expression level of the mRNA gene of the inflammatory factor of the composition horny layer cell: the method is the same as in example 2
3. Results
The results are shown in tables 3-5 below:
it can be seen from tables 3 to 4 that, with respect to the antibacterial effect against Staphylococcus epidermidis, polygonum cuspidatum has a significant effect on the antibacterial result (p < 0.05). From the extreme difference and extreme value analysis, the concentration of giant knotweed is 1.5%, the concentration of stemona is 1-1.5%, the concentration of pseudo-ginseng extract is 1% and the concentration of rice fermentation extract is more than 2%, when combined compatibility is carried out, the antagonistic activity of staphylococcus epidermidis is strongest.
As can be seen from tables 3 to 5, the extract of Polygonum cuspidatum, panax notoginseng and fermentation showed a significant effect (p < 0.05) on inhibiting the expression of keratinocyte inflammatory factor. From the extremely poor and extreme value analysis, the concentration of giant knotweed is 0.5-1.0%, the concentration of stemona is 1.5-2.0%, the concentration of pseudo-ginseng extract is 1.5-2.0% and the concentration of rice fermentation extract is more than 1-2%, and the effect of inhibiting the expression of keratinocyte inflammatory factor is best when the combination compatibility is carried out.
In combination with the above analysis, we finally selected the following combinations for further validation experiments.
As can be seen from Table 6 above, the four combinations of the new combinations 1 to 4 obtained all showed good effects of antagonizing Staphylococcus epidermidis and inhibiting the expression of inflammatory factors in keratinocytes.
From the orthogonal results, the radix stemonae extract has no obvious influence on bacteriostasis and anti-inflammatory. In view of this, activity studies were performed without the addition of stemona extract. However, the results show (see Table 6 for new compositions A-J): the antibacterial rate of the composition is reduced under the condition of no stemona root addition; and the expression of inflammatory factors is synchronously improved. But the addition amount of the stemona exceeds 2%, the comprehensive effect is not obviously improved.
Showing that Stemona has synergistic effect on inhibiting bacteria and relieving inflammatory reaction with other components in the composition.
From the condition of adding giant knotweed, the antibacterial rate is greatly reduced under the condition of no giant knotweed; the expression of inflammatory factor gene is also obviously improved, which indicates that the antibacterial effect and anti-inflammatory response may have important connection. However, if the addition amount of the stemona root exceeds 2%, the comprehensive effect is not obviously improved (see the novel compositions A-J in Table 6).
From the condition of adding the pseudo-ginseng extract, the antibacterial rate is greatly reduced under the condition of no addition; the expression of inflammatory factor gene is also obviously improved, which again shows that the antibacterial substance and the anti-inflammatory substance have synergistic effect. But the addition amount of the pseudo-ginseng reaches 4 percent, so that the comprehensive effect is not obviously improved. After more than 4% the combined system became unstable and a precipitate appeared (see Table 6 for new compositions A-J).
From the addition of the rice fermentation extract, the whole combined system is unstable without addition. The brown yellow precipitate is easy to separate out. After exceeding 3%, the comprehensive effect is not improved obviously. After more than 6%, the odor of the combined system was not coordinated (see Table 6 for new compositions A-J).
By combining the above results, the various components are cooperated with each other, and the addition amount is limited in a specific range.
Example five, combined cytotoxicity and eye irritation test of the invention
1. Experimental materials and reagents: the 6 combinations in example four (Table 6) were taken.
Method
2.1 neutral Red cell toxicity test method and test results
Neutral red is a weak cationic dye that is extremely susceptible to penetrating the cell membrane and aggregating in the cell lysosomes in a non-ionic diffusion manner. If the change of the cell surface or the change of the sensitivity of the lysosome membrane causes irreversible changes such as increased lysosome fragility, the ability of the cell to absorb and link neutral red is reduced. This distinguishes normal, injured, dead cells. The principle of detection of the cytotoxicity neutral red absorption method is that after a test object contacts 3T3 cells, the cell activity is observed, and the IC of the test object is calculated 50 (the concentration of the test substance that decreases the cell activity by 50%). LD is then calculated by predictive model for acute oral toxicity 50 (half lethal dose). The acute oral toxicity grading of the test object is obtained by GHS chemical acute toxicity grading。
2.2 Eye irritation test (CAMVA test) method and test result
Chorioallantoic membrane vascular test (Chorioallantoic membrane vascular assay, CAMVA) procedure the potential ocular irritation was assessed by observing the effect of the test on blood vessels, taking 14-day-old chick embryos, the test directly contacting small areas of the CAM, and observing changes in the CAM blood vessels, such as bleeding or congestion (telangiectasia), and vessel disappearance (ghosting), after 30 minutes exposure, such that the concentration of the test at which 50% of fertilized eggs develop these lesions was considered as the toxic endpoint. The CAMVA test requires the determination of the average integral of positive response in 10 chick embryos tested. It is generally believed that the test subjects do not respond to stimulation when the average integral of the CAMVA test is < 1.0.
Results
The results are shown in tables 7 and 8 below:
the test results in table 7 show that the safety of the anti-dandruff compositions is effectively ensured, the toxicity level of the anti-dandruff compositions is fifth grade, and the toxicity level is obviously lower than that of various common cosmetic additives (generally, the toxicity level is fourth grade). These anti-dandruff compositions are therefore not subject to an increase in their toxicity.
Table 8 shows that the average score of CAMVA test of the anti-dandruff composition is less than 1.0, thus the anti-dandruff composition does not cause potential irritation or allergic response to the human body.
Example 5: application experiment of volunteers of the invention
Specific case 1: su somewhere, man, 28 years old. Dandruff is very severe. The effect is poor by using commercial anti-dandruff shampoo. After the traditional Chinese medicine composition provided by the invention is used for washing hair, after the traditional Chinese medicine composition is used for 3 times, the dandruff condition is greatly relieved, the shampoo is not washed for one week, and the dandruff is less (the comparison between the front and the rear of the anti-dandruff composition is shown in figure 1).
Specific case 2: forest somebody, male, 41 years old. Has intractable scalp, and scalp itching. After the traditional Chinese medicine composition provided by the invention is used for washing hair, after the traditional Chinese medicine composition is used for 3 times a week, the dandruff condition is controlled, and after the traditional Chinese medicine composition is used for 3 months, the dandruff condition is greatly relieved (the comparison photo of the anti-dandruff composition before and after the anti-dandruff composition is used is shown in figure 2).
Specific case 3: tang somewhere, woman, 43 years old. Has more obvious dandruff. The shampoo should be used for washing hair every day, and the effect of using the commercially available anti-dandruff shampoo is not obvious. The traditional Chinese medicine composition provided by the invention is used for washing hair, and after the traditional Chinese medicine composition is used for 1 week, the dandruff condition is relieved. (see FIG. 3 for photographs of the anti-dandruff composition of the invention before and after use).
Finally, it is noted that the above-mentioned preferred embodiments illustrate rather than limit the invention, and that, although the invention has been described in detail with reference to the above-mentioned preferred embodiments, it will be understood by those skilled in the art that various changes in form and details may be made therein without departing from the scope of the invention as defined by the appended claims.
Claims (4)
1. A traditional Chinese medicine anti-dandruff composition is characterized by comprising the following components in parts by weight: 1.0 to 3.0 percent of giant knotweed extract, 1.5 to 2.5 percent of Baistep extract, 1.5 to 4.0 percent of pseudo-ginseng extract, 2.0 to 3.0 percent of rice ferment and the balance of process water.
2. A method of preparing a composition according to claim 1, comprising the steps of:
s1, preparation of an extract: the traditional Chinese medicine materials are prepared by the following steps: respectively placing rhizoma Polygoni Cuspidati, radix et rhizoma Rhei, notoginseng radix and rice yeast in percolating tank, adding 70% ethanol 5 times of the weight of the materials, soaking at room temperature for 48 hr, and percolating to obtain extractive solution; adding 60% ethanol with 5 times of the weight of the medicinal materials into a percolation tank respectively, soaking for 48 hours, percolating to obtain extract, mixing the two extracts, and uniformly mixing to obtain extract of different Chinese medicinal materials;
s2, concentrating an extracting solution: at the temperature: 72.+ -. 1 ℃ and vacuum degree: -0.05-0.06 mpa, rotational speed: 30+/-1 rpm, adding the uniformly mixed extracting solution into a concentrating device, and concentrating to 30% of the initial mass to obtain concentrated solutions of all components;
s3, blending: stirring and heating the concentrated solutions of all the components to 80-85 ℃ at the rotating speed of 40+/-5 rpm, adding the mixed solution into a wide-mouth container proportionally, mixing, heating in a water bath, keeping the temperature at 80-85 ℃, preserving heat for 1h in an open way, and continuously stirring during the heating and heat preserving processes to concentrate the concentrated solution to 50% of the mass during the adding;
s4, stirring and mixing uniformly after heat preservation is finished, and quantifying the process water to 100% to obtain the traditional Chinese medicine anti-dandruff composition.
3. Use of a composition according to claim 1 in an external preparation for the prevention and treatment of dandruff or in a scalp care product.
4. Use of a composition according to claim 1 in a product for inhibiting staphylococcus epidermidis, reducing scalp inflammation.
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