CN112089652A

CN112089652A - Compound traditional Chinese medicine bacteriostatic composition and preparation method and application thereof

Info

Publication number: CN112089652A
Application number: CN202010843114.8A
Authority: CN
Inventors: 宋丽雅; 何聪芬; 杨雪晨; 崔树梅; 贾焱; 成志伟; 董坤; 刘蕾
Original assignee: Beijing Technology and Business University
Current assignee: Beijing Technology and Business University
Priority date: 2020-08-20
Filing date: 2020-08-20
Publication date: 2020-12-18

Abstract

The invention discloses a compound traditional Chinese medicine bacteriostatic composition and a preparation method and application thereof. The compound traditional Chinese medicine bacteriostatic composition consists of a schisandra chinensis extracting solution, a myrobalan extracting solution, a raw sanguisorba extracting solution and water. Preferably, the volume ratio of the schisandra chinensis extracting solution, the myrobalan extracting solution, the raw sanguisorba extracting solution and water is 2:3:1: 3. The compound traditional Chinese medicine bacteriostatic composition is prepared by mixing the schisandra chinensis extracting solution, the myrobalan extracting solution and the raw sanguisorba extracting solution, the bacteriostasis of the traditional Chinese medicine bacteriostatic composition is increased by compounding the three traditional Chinese medicine extracting solutions, the use amount of a single traditional Chinese medicine is reduced, the growth of microorganisms can be effectively inhibited by adding the traditional Chinese medicine bacteriostatic composition into cosmetics, and the traditional Chinese medicine bacteriostatic composition has the characteristics of wide bacteriostatic range and less adverse reaction. Therefore, the compound traditional Chinese medicine bacteriostatic composition has a good application prospect.

Description

Compound traditional Chinese medicine bacteriostatic composition and preparation method and application thereof

Technical Field

The invention relates to a compound traditional Chinese medicine composition with an antibacterial effect, and a preparation method and application thereof, and belongs to the technical field of traditional Chinese medicines.

Background

Currently, as preservatives used in cosmetics in many cases, parabens, formaldehyde-releasing agents, a mixture of methylchloroisothiazolinone and methylisothiazolinone (CMIT/MIT), Methylisothiazolinone (MIT), iodopropynyl butylcarbamate (IPBC), and the like are used. Although the chemical preservatives have a certain inhibiting effect on microorganisms, the preservatives also have certain potential side effects and potential harm to human health, so that natural preservatives which are convenient to develop and use and have better safety are generally regarded by countries all over the world.

In China, with the vigorous national advance of the development of the traditional Chinese medicine industry, traditional Chinese medicines are widely used in foods and cosmetics. Natural bacteriostatic agents (also called natural bacteriostatic agents) of plant origin are preferred by people because of their advantages of natural non-toxicity, high safety and the like. In recent years, people continuously discover new bacteriostatic plants with higher application value through bacteriostatic test research and multidisciplinary comprehensive research, so that bacteriostatic plant resources are continuously enriched and enriched. The application of the natural plant component extracting solution as the bacteriostatic agent meets the pursuit of natural and non-chemical raw materials of consumers and has other effects at the same time.

In view of the above, the invention provides a compound traditional Chinese medicine bacteriostatic composition prepared by compounding a schisandra chinensis extracting solution, a myrobalam extracting solution and a raw sanguisorba extracting solution, and the bacteriostatic composition has the characteristics of wide bacteriostatic range and less adverse reaction.

Disclosure of Invention

The invention aims to solve the technical problem of providing a bacteriostatic composition developed from traditional Chinese medicines, which has the characteristics of high safety, less adverse reaction, strong bacteriostatic effect and unique function and effect.

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

the invention obtains seven traditional Chinese medicines, namely schisandra chinensis, polygonum cuspidatum, myrobalan, raw sanguisorba, vine tea, suberect spatholobus stem and fructus psoraleae, from a plurality of traditional Chinese medicines with bacteriostatic action through preliminary screening. The antibacterial activity of the seven traditional Chinese medicine extracts is further researched, the traditional Chinese medicine extracts with strong antibacterial activity are compounded and formulated, the antibacterial activity of the formulation is measured, and the seven traditional Chinese medicine extracts are added into cosmetics for corrosion prevention challenge. The results show that the Chinese magnoliavine fruit/myrobalan/raw sanguisorba officinalis extract compounded traditional Chinese medicine bacteriostatic agent has synergistic and additive bacteriostatic effects, and meanwhile, the compounded formula has the characteristics of low minimum bacteriostatic concentration and wide bacteriostatic spectrum.

Therefore, on the basis of the research, the invention provides a compound traditional Chinese medicine bacteriostatic composition which is composed of a schisandra chinensis extracting solution, a myrobalan extracting solution, a raw sanguisorba extracting solution and water.

Wherein fructus Schisandrae is dried mature fruit of Schisandra chinensis (Turcz.) Baill of Magnoliaceae, and has effects of astringing, arresting discharge, invigorating qi, promoting fluid production, invigorating kidney, and calming heart. Can be used for treating chronic cough, asthma, nocturnal emission, enuresis, frequent micturition, chronic diarrhea, spontaneous perspiration, night sweat, thirst due to body fluid consumption, internal heat, diabetes, palpitation, and insomnia. The schisandra contains various chemical components, and literature reports mainly include lignans, polysaccharides, volatile oil, organic acids and the like, wherein the content of the lignans is the highest and can reach 8%. The fructus Schisandrae chinensis extractive solution has antiinflammatory, free radical scavenging, lipid peroxidation resisting, and immunity enhancing effects. At present, schisandra chinensis extract is applied to cosmetics as an effective component. Researches show that the schisandra extract has high heat stability and antibacterial activity on bacteria such as escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and the like.

Myrobalan is a commonly used medicinal material of multiple nationalities, is widely applied to Mongolian medicines, traditional Chinese medicines and Tibetan medicines, and is also widely applied to traditional medicines of southeast Asia countries such as India and Iran. Fructus Chebulae contains various active ingredients such as tannin, phenolic acid, triterpene, polysaccharide, and volatile oil. Modern pharmacological studies show that the myrobalan has the effects of resisting atherosclerosis, relieving diarrhea, protecting liver and benefiting gallbladder, resisting bacteria, resisting mutagenesis and the like. In addition, the myrobalan has certain bacteriostatic action on pseudomonas aeruginosa, streptococcus agalactiae, escherichia coli and the like.

Raw Sanguisorba officinalis is derived from dried root of Sanguisorba officinalis L. or Sanguisorba longata L. var. longifolia (Bert.) Yu et Li, which is a Rosaceae plant. It is slightly cold in nature, bitter, sour and astringent in flavor, and enters liver and large intestine meridians. Has effects of cooling blood, stopping bleeding, removing toxic substance, and healing sore. Researches show that sanguisorba contains abundant nutrients and physiological active ingredients, mainly tannin, saponin and flavonoid, particularly abundant tannin. The radix Sangusorbae extractive solution has certain antibacterial effect on typhoid bacillus, dysentery bacillus, Streptococcus mutans, viscous actinomyces, and Lactobacillus acidophilus.

Preferably, the schisandra chinensis extracting solution is prepared by the following method: air drying and crushing Chinese magnoliavine fruit serving as a traditional Chinese medicine raw material, sieving with a 100-mesh sieve, mixing with distilled water according to a material-liquid ratio of 1:10, standing for 30 minutes after water bath, pouring liquid into a centrifuge tube, centrifuging at a high speed, and performing rotary evaporation and concentration on supernatant to obtain the Chinese magnoliavine fruit extracting solution.

Preferably, the myrobalan extract is prepared by the following method: air-drying and crushing traditional Chinese medicine raw material myrobalan, sieving with a 100-mesh sieve, mixing with distilled water according to a material-liquid ratio of 1:10, standing for 30 minutes after water bath, pouring liquid into a centrifuge tube, centrifuging at high speed, taking supernatant, and performing rotary evaporation and concentration to obtain the myrobalan extract.

Preferably, the sanguisorba officinalis extracting solution is prepared by the following method: air drying and crushing raw sanguisorba officinalis which is a traditional Chinese medicine raw material, sieving by a 100-mesh sieve, mixing with distilled water according to a material-liquid ratio of 1:10, standing for 30 minutes after water bath, pouring liquid into a centrifuge tube, centrifuging at a high speed, taking supernate, and carrying out rotary evaporation and concentration to obtain the raw sanguisorba officinalis extracting solution.

Preferably, the high-speed centrifugation is performed for 5min at 8000 r/min.

Preferably, the schisandra chinensis extract, the myrobalan extract and the raw sanguisorba officinalis extract are concentrated according to the same volume percentage.

Preferably, the volume ratio of the schisandra chinensis extracting solution, the myrobalan extracting solution, the raw sanguisorba extracting solution and water is 2:3:1: 3.

Furthermore, the invention also provides application of the compound traditional Chinese medicine bacteriostatic composition in inhibiting bacterial growth and proliferation.

Preferably, the compound traditional Chinese medicine bacteriostatic composition has the effect of inhibiting growth and proliferation of escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and candida albicans.

Compared with the traditional bacteriostatic agent, the bacteriostatic agent has the advantages that:

the traditional Chinese medicine bacteriostatic composition is prepared by mixing traditional Chinese medicine extract, is a traditional Chinese medicine compound natural bacteriostatic agent, has the characteristics of wide bacteriostatic range and less adverse reaction, can inhibit the growth of undetected bacteria in cosmetics, and has the functions of inflammation resistance and oxidation resistance due to multiple effective components of traditional Chinese medicines. Therefore, the compound traditional Chinese medicine bacteriostatic composition can be used in cosmetics and has a good application prospect.

Detailed Description

The present invention is further illustrated by the following specific examples, but it should be understood that these examples and experimental examples are intended only for the purpose of more detailed description, and are not intended to limit the present invention in any way. It will be apparent to those skilled in the art that the materials and methods of operation used in the present invention are well known in the art, unless otherwise specified.

Example 1

1. Materials and methods

1.1 Experimental materials and instruments

1.1.1 Experimental strains

Escherichia coli (Escherichia coli), Staphylococcus aureus (Staphylococcus aureus), Candida albicans (Candida albicans), Pseudomonas aeruginosa (Pseudomonas aeruginosa)

1.1.2 subjects

Fructus Schisandrae chinensis, rhizoma Polygoni Cuspidati, fructus Chebulae, radix Sangusorbae, Ampelopsis grossedentata, caulis Spatholobi, and fructus Psoraleae.

1.1.3 laboratory instruments and apparatus

TABLE 1 Experimental instruments and apparatus

1.2 Experimental methods

1.2.1 preparation of Chinese medicine extract

Air drying and pulverizing Chinese medicinal materials (fructus Schisandrae chinensis, rhizoma Polygoni Cuspidati, fructus Chebulae, radix Sangusorbae, folium Ampelopsis Grossdentata, caulis Spatholobi or fructus Psoraleae), sieving with 100 mesh sieve, extracting with water to obtain 50g Chinese medicinal powder, mixing with 500ml distilled water (material-liquid ratio of 1:10), water bathing at 50 deg.C above, and standing for 30 min. Pouring the liquid into a centrifuge tube, centrifuging at high speed (v 8000r/min, t 5min), taking the supernatant, and concentrating by rotary evaporation to 30% of the original volume to obtain seven Chinese medicinal extractive solutions.

1.2.2 Chinese medicinal extractive solution and its formula antibacterial activity

(1) Oxford cup bacteriostasis test

The method comprises the steps of inhibiting the growth of microorganisms in a culture medium to form an inhibition zone by utilizing the diffusion of an object to be detected in the culture medium, visually comparing the inhibition capacity of the object to be detected according to the inhibition zone, and measuring the inhibition activity of seven traditional Chinese medicine extracting solutions on four bacteria by taking staphylococcus aureus, escherichia coli, candida albicans and pseudomonas aeruginosa as experimental strains.

The experimental steps are as follows: preparing culture medium, sterilizing all the materials and solution (121 deg.C, 20min), sterilizing solid culture medium, cooling to about 50 deg.C, and pouring into flat plate. Inoculating the single colony into a liquid culture medium, and placing the liquid culture medium into a shaking table for culturing bacteria for 24 hours and fungi for 48 hours for later use. Diluting part of the Chinese medicinal extractive solution to different concentrations (concentration after rotary evaporation)&Dilute concentration). 200 μ L of the bacterial suspension was centrifuged, and the supernatant was decanted. Resuspending the solid pellet and measuring the OD (bacterial OD)₆₂₅0.09 fungal OD₆₂₅0.7), the solution concentration was 10⁸CFU/mL. The solution was diluted 100-fold with physiological saline (i.e., 10 ×)⁶CFU/mL). A100. mu.L plate was coated with the solution. Three Oxford cups (240. mu.L each) were placed per medium and the herbal extract was added. Placing the mixture into a microorganism incubator (bacteria 37 ℃, fungi 28 ℃) to shake and culture for 24 hours (fungi is more than 48 hours) and observing the bacteriostasis (whether a bacteriostasis zone exists and the size of the bacteriostasis zone).

(2) Determination of Minimum Inhibitory Concentration (MIC)

And adding a series of traditional Chinese medicine extracting solutions with different concentrations into bacterial liquid with the same concentration and volume for culturing by using a sesquidilution method, wherein the minimum concentration of the liquid medicine for inhibiting the bacterial growth is the minimum bacteriostatic concentration of the tested medicine.

The experimental steps are as follows: preparing a liquid culture medium and a solid culture medium, sterilizing all tools (Ep tubes, gun heads, inoculating rings and the like) and the culture medium, pouring a flat plate after the solid culture medium is cooled, inoculating 5 microorganisms into the liquid culture medium, putting the liquid culture medium into a constant-temperature shaking table for shake culture, streaking bacteria for 24 hours and fungi for 48 hours on the solid culture medium, and putting the solid culture medium into an incubator for continuous culture. After the same time, the single colony on the solid culture medium is inoculated with normal saline to determine the concentration, and 10 is obtained⁶CFU/mL, diluting the extractive solution for 10 times to obtain 11 medicinal liquids with different concentrations, and mixing 500 μ L of the extractive solutions with different concentrations and 500 μ LAnd mixing the L bacterial liquids, putting the mixture into an incubator for culture, and observing the result after 24h/28 h.

(3) Experiment of corrosion prevention challenge

Adding a certain amount of microorganisms into the cosmetics and periodically detecting the growth condition of the microorganisms so as to judge the preservative property of the cosmetics. The experimental steps are as follows: the emulsion was prepared in a 100mL system.

TABLE 2 emulsion formulation

The method is described in United states Pharmacopeia, 21 st edition, and bacteria are mixed after culturing to a final concentration of 5 × 10⁶CFU/mL, preparation of mixed fungi 3X 10⁵CFU/mL, weighing the sample, adding one mixed strain per 30g, fully mixing, and placing at room temperature. The test was performed every 0, 7, 14, 21, 28 days. On the test day, 0.3g of sample was taken, added to a sterilized Ep tube containing glass beads, 2.7mL of sterilized normal saline was added, and demulsification was performed by shaking. Then diluting twice with normal saline 10 times to obtain gradient diluted sample, coating and counting the bacteria content in the sample with flat plate, culturing the bacteria at 37 deg.C for 24h, and culturing the fungi at 28 deg.C for 48 h.

The samples contained bacteria at day 28>10³CFU/g (mL). The sample fails to pass a microorganism experiment, which shows that the sample cannot effectively inhibit microorganisms and is easy to be polluted. The samples contained bacteria at 10 days 28²-10³CFU/g (mL), the sample is qualified by a microorganism experiment, and the preservative system is feasible when the protein or other animal and plant components in the product are not high, the production environment meets the sanitary standard, and the package is not easy to cause secondary pollution.

The sample contains bacteria at 10-10 days 28²CFU/g (mL). The sample shows that the antiseptic system has strong inhibition effect on microorganisms through microorganism experiments, and is not easily polluted during production, storage and use of productsAnd (6) dyeing.

2. Results and analysis

2.1 Single herb bacteriostatic activity of Chinese herb extract

An oxford cup method is utilized to carry out bacteriostasis experiments on a plurality of microorganisms, DMSO is selected to dilute MTI to the same concentration to be used as a positive control, and water is used as a negative control. The mean and standard deviation of the diameters of the three oxford rings on each dish were calculated.

TABLE 3 bacteriostatic activity (D/mm) of different herbal extracts

As shown in the table 3, the extract of Chinese magnoliavine fruit with stronger antibacterial activity to escherichia coli has the diameter of an inhibition zone of 14.16 +/-1.4 mm and belongs to moderate sensitivity (D is more than 10mm and less than 15 mm); the strong bacteriostatic activity to staphylococcus aureus is that raw sanguisorba extract (D is 28.95 +/-2.32 mm), caulis spatholobi extract (D is 25.85 +/-1.55 mm), schisandra extract (D is 23.55 +/-0.39 mm), vine tea extract (D is 22.49 +/-0.77 mm) and myrobalan extract (D is 20.56 +/-1.36 mm) are all extremely sensitive (D is more than 20 mm); the antibacterial activity to pseudomonas aeruginosa is stronger than that of schisandra extract (D is 18.29 +/-0.99 mm) and myrobalan extract (D is 18.17 +/-1.06 mm), and the extract is highly sensitive (D is more than 15mm and less than 20 mm); the most powerful bacteriostatic activity against candida albicans is myrobalan (D is 22.41 +/-1.06 mm), and the myrobalan is extremely sensitive (D is more than 20 mm). In conclusion, of the seven traditional Chinese medicine extracting solutions, the schisandra extracting solution has better bacteriostatic effect on escherichia coli and pseudomonas aeruginosa, the myrobalan extracting solution has better bacteriostatic effect on candida albicans and pseudomonas aeruginosa, and the sanguisorba officinalis extracting solution has better bacteriostatic effect on staphylococcus aureus, so that the three extracting solutions are selected to carry out pairwise compound bacteriostatic activity test.

2.2 bacteriostatic activity of two compound traditional Chinese medicine extracts

The sanguisorba officinalis extract and the schisandra chinensis extract, the sanguisorba officinalis extract and the myrobalan extract, the schisandra chinensis extract and the myrobalan extract are compounded in equal volume respectively, and the sanguisorba officinalis extract, the schisandra chinensis extract and the myrobalan extract are mixed with water in equal volume respectively to serve as a control group. DMSO was chosen to dilute the MTI to the same concentration as the positive control, water as the negative control.

TABLE 4 antibacterial activity of two drugs (D/mm)

As can be seen from the above table 4, the three compound compositions have the highest antibacterial activity on escherichia coli, namely the compound of the schisandra chinensis and the myrobalan extract (9.5 +/-0.53 mm) is slightly improved in antibacterial activity compared with the single extract; the highest bacteriostatic activity to staphylococcus aureus is that the extract of the myrobalan and the raw sanguisorba is compounded (28.07 +/-1.88 mm), and the bacteriostatic activity is slightly improved compared with that of a single extract; the highest bacteriostatic activity on pseudomonas aeruginosa is that the compound of the schisandra chinensis and the myrobalan extract (24.31 +/-1.02 mm) and the compound of the schisandra chinensis and the raw sanguisorba extract (22.19 +/-0.37 mm) are slightly improved compared with the single extract; the best bacteriostatic activity to candida albicans is that the myrobalan and raw sanguisorba officinalis extract is compounded (27.68 +/-1.39 mm), and the bacteriostatic activity is slightly improved compared with that of a single extract. Experimental results show that the antibacterial activity of the two extracting solutions after compounding is improved, and mutual inhibition is avoided. In order to further improve the bacteriostatic activity of the formula and expand the bacteriostatic spectrum of the formula, the schisandra chinensis, the raw sanguisorba officinalis and the myrobalan extracting solution are compounded.

2.3 bacteriostatic activity of three-medicine compound of traditional Chinese medicine extract

2.3.1 Quadrature compounding

In order to obtain the optimal ratio of the extracts of schisandra, raw sanguisorba and myrobalan, the three-factor four-level orthogonal compounding is adopted, the three-factor four-level extract of schisandra, raw sanguisorba and myrobalan is adopted, and the four levels are four different volumes (table 5).

TABLE 5 Combined Quadrature Table

As can be seen from the orthogonal table, there are 16 possible concentration combinations for each of the three combinations.

2.3.2 results of orthogonal experiments

TABLE 6 inhibition zone (D/mm) of compound formula of fructus Schisandrae chinensis, fructus Chebulae and radix Sangusorbae extract on Escherichia coli

As shown in Table 6, in the experiment of inhibiting bacteria of Escherichia coli by the combination of the extracts of fructus Schisandrae chinensis, fructus Chebulae and radix Sangusorbae, R is_B＞R_C＞R_ATherefore, the largest factor influencing the bacteriostatic activity of the formula is B, then C and finally A, according to the K values of the three substances, the optimal formula compounded by the schisandra chinensis, the myrobalan and the raw sanguisorba extract is A4C1A4, and the formula with better bacteriostatic effect on escherichia coli is the formula of the schisandra chinensis and the myrobalan extract in consideration of water used for constant volume, and the final volume ratio of the schisandra chinensis extract, the myrobalan extract and the water is 3:3: 3.

TABLE 7 inhibition zone (D/mm) of compound formula of fructus Schisandrae chinensis, fructus Chebulae and radix Sangusorbae extract on Staphylococcus aureus

As shown in Table 7, in the bacteriostatic experiment of the compound formula of the extracts of the schisandra chinensis, the myrobalan and the raw sanguisorba officinalis on staphylococcus aureus, R is_A＞R_B＞R_CTherefore, the largest factor influencing the bacteriostatic activity of the formula is A, then B and finally C, and according to the K values of the three substances, the optimal formula compounded by the schisandra chinensis, the raw sanguisorba and the myrobalan extracting solution is A4B4C2, namely the final volume ratio of the schisandra chinensis extracting solution, the myrobalan extracting solution, the raw sanguisorba extracting solution and water is 3:3:1: 2.

TABLE 8 antibacterial zone (D/mm) of compound formula of fructus Schisandrae chinensis, fructus Chebulae and radix Sangusorbae extract on Pseudomonas aeruginosa

As shown in Table 8, in the bacteriostasis experiment of the combination of the extracts of the schisandra chinensis, the myrobalan and the raw sanguisorba officinalis on pseudomonas aeruginosa, R is_B＞R_A＞R_CTherefore, the largest factor influencing the bacteriostatic activity of the formula is B, then A and finally C, and according to the K values of the three substances, the optimal formula compounded by the schisandra chinensis, the raw sanguisorba and the myrobalan extracting solution is B4A3C2, namely the final volume ratio of the schisandra chinensis extracting solution, the myrobalan extracting solution, the raw sanguisorba extracting solution and water is 2:3:1: 3.

TABLE 9 antibacterial zone (D/mm) of compound formula of fructus Schisandrae chinensis, fructus Chebulae and radix Sangusorbae extract on Candida albicans

As shown in Table 9, in the Candida albicans inhibition experiment of the combination of the extracts of fructus Schisandrae chinensis, fructus Chebulae and radix Sangusorbae, R_A＞R_B＞R_CTherefore, the largest factor influencing the bacteriostatic activity of the formula is A, then C and finally B, and according to the K values of the three substances, the optimal formula compounded by the fructus schizandrae, the myrobalan and the raw sanguisorba extract is A1B4C4, namely the volume ratio of the myrobalan extract to the raw sanguisorba extract to water is 1:1: 1.

In conclusion, the schisandra chinensis, the myrobalan and the raw sanguisorba can be prepared into four formulas with different concentrations, namely a formula I: compounding the schisandra chinensis extracting solution, the myrobalan extracting solution and water according to the volume ratio of 1:1: 1; and the formula II comprises: compounding the schisandra chinensis extracting solution, the myrobalan extracting solution, the raw sanguisorba officinalis extracting solution and water according to the volume ratio of 3:3:1: 2; compounding a schisandra chinensis extracting solution, a myrobalan extracting solution, a raw sanguisorba officinalis extracting solution and water according to a volume ratio of 2:3:1: 3; and fourthly, compounding the myrobalan extract, the raw sanguisorba extract and water according to the volume ratio of 1:1: 1.

2.3.3 determination of bacteriostatic activity of two compound compositions

The antimicrobial activity of the four formulations on four bacteria was determined by oxford cup method, and the experimental results are shown in table 10.

TABLE 10 bacteriostatic Activity of four formulations (D/mm)

The results in the table 10 show that the third formula has high sensitivity (D is more than 15mm and less than 20mm) to staphylococcus aureus and candida albicans, and has low sensitivity (D is more than 10mm and less than 15mm) to escherichia coli and pseudomonas aeruginosa; the formula has moderate sensitivity (D is more than 11mm and less than 15mm) to staphylococcus aureus and candida albicans, but has low sensitivity (D is less than 10mm) to escherichia coli and pseudomonas aeruginosa; but the second formula has no sensitivity to candida albicans and the fourth formula has no sensitivity to escherichia coli. From the consideration of antibacterial spectrum of the formula after compounding, a formula I and a formula III are screened out for application research. In order to confirm the antibacterial activity of the formula and determine the dosing concentration after the cosmetics are prepared, the Minimum Inhibitory Concentration (MIC) values of the first formula, the third formula and the single extracting solution are further tested, and the interaction effect of each component in the compound formula is judged.

2.3.4 determination of the Minimum Inhibitory Concentration (MIC)

In order to determine the antibacterial activity before and after the compounding of the extracting solutions of schisandra chinensis, myrobalan and raw sanguisorba officinalis, a formula I is as follows: compounding the schisandra chinensis extracting solution, the myrobalan extracting solution and water according to the volume ratio of 1:1: 1; the formula III is as follows: compounding the schisandra chinensis extracting solution, the myrobalan extracting solution, the raw sanguisorba officinalis extracting solution and water according to the volume ratio of 2:3:1: 3; the stock solutions of fructus Schisandrae chinensis extractive solution, fructus Chebulae extractive solution and radix Sangusorbae extractive solution are collectively marked as C₀. The minimum inhibitory concentrations for the three individual drugs and the two formulations are shown in table 11.

As can be seen from Table 11, the MICs for a pair of E.coli, Staphylococcus aureus, Pseudomonas aeruginosa and Candida albicans in the formulation are 1/16C respectively₀，1/16C₀，1/16C₀And 1/8C₀The formula shows that the composition has better bacteriostatic action on escherichia coli, staphylococcus aureus and pseudomonas aeruginosa. Three pairs of colibacillus, staphylococcus aureus and pseudomonas aeruginosaAnd the MIC of Candida albicans is 1/8C respectively₀，1/16C₀，1/16C₀And<1/16C₀the third formula has stronger bacteriostatic action on staphylococcus aureus, pseudomonas aeruginosa and candida albicans. Compared with the two formulas, the compound product of the three formulas has better effect on bacteria and fungi. Therefore, the third formula is applied to the cosmetics, and a preservation challenge experiment is carried out.

TABLE 11 minimum inhibitory concentrations of the combination formulas on four bacteria

2.4 Corrosion protection challenge

2.4.1 emulsion preparation

Preparing emulsion according to the finally obtained compound formula III (the schisandra extract, the myrobalan extract, the raw sanguisorba extract and water are compounded according to the volume ratio of 2:3:1: 3) and the measured MIC according to a formula table, and respectively adding a compound formula of 4MIC and 1 MIC.

2.4.2 Corrosion protection challenges

As shown in Table 12, the inhibition ability of the third pair of bacteria is good in 28-day preservation challenge (the number of colonies on day 28 is less than 10), and the third pair of bacteria has strong bacteriostatic activity and is not easy to be polluted in the production, storage and use processes of the product through a microorganism challenge experiment.

TABLE 12 microbial (bacteria) preservation challenge test results Table (colony number)

3 conclusion and prospect

The Chinese medicinal bacteriostatic composition compounded by the extracts of the schisandra chinensis, the myrobalan and the raw sanguisorba has stronger synergistic and additive bacteriostatic effects and has the characteristics of low minimum bacteriostatic concentration and wider bacteriostatic spectrum. The schisandra chinensis, myrobalan and raw sanguisorba officinalis extract are compounded, so that the antibacterial activity of the compound is increased, the use amount of single traditional Chinese medicine is reduced, and the compound can effectively inhibit the growth of microorganisms when added into cosmetics. The invention has guiding significance for the research of the traditional Chinese medicine bacteriostatic agent.

Claims

1. A compound traditional Chinese medicine bacteriostatic composition is characterized by comprising a schisandra chinensis extracting solution, a myrobalan extracting solution, a raw sanguisorba officinalis extracting solution and water.

2. The compound traditional Chinese medicine bacteriostatic composition according to claim 1, wherein the schisandra chinensis extracting solution is prepared by the following method: air drying and crushing Chinese magnoliavine fruit serving as a traditional Chinese medicine raw material, sieving with a 100-mesh sieve, mixing with distilled water according to a material-liquid ratio of 1:10, standing for 30 minutes after water bath, pouring liquid into a centrifuge tube, centrifuging at a high speed, and performing rotary evaporation and concentration on supernatant to obtain the Chinese magnoliavine fruit extracting solution.

3. The compound traditional Chinese medicine bacteriostatic composition according to claim 1, wherein the myrobalan extract is prepared by the following method: air-drying and crushing traditional Chinese medicine raw material myrobalan, sieving with a 100-mesh sieve, mixing with distilled water according to a material-liquid ratio of 1:10, standing for 30 minutes after water bath, pouring liquid into a centrifuge tube, centrifuging at high speed, taking supernatant, and performing rotary evaporation and concentration to obtain the myrobalan extract.

4. The compound traditional Chinese medicine bacteriostatic composition as claimed in claim 1, wherein the sanguisorba officinalis extracting solution is prepared by the following method: air drying and crushing raw sanguisorba officinalis which is a traditional Chinese medicine raw material, sieving by a 100-mesh sieve, mixing with distilled water according to a material-liquid ratio of 1:10, standing for 30 minutes after water bath, pouring liquid into a centrifuge tube, centrifuging at a high speed, taking supernate, and carrying out rotary evaporation and concentration to obtain the raw sanguisorba officinalis extracting solution.

5. The compound traditional Chinese medicine bacteriostatic composition according to any one of claims 2 to 4, wherein the high-speed centrifugation is performed at 8000r/min for 5 min.

6. The compound traditional Chinese medicine bacteriostatic composition according to any one of claims 2 to 4, wherein the schisandra chinensis extracting solution, the myrobalan extracting solution and the raw sanguisorba extracting solution are concentrated according to the same volume percentage.

7. The compound traditional Chinese medicine bacteriostatic composition according to claim 1, wherein the volume ratio of the schisandra chinensis extracting solution, the myrobalan extracting solution, the raw sanguisorba extracting solution and water is 2:3:1: 3.

8. The use of the compound traditional Chinese medicine bacteriostatic composition of any one of claims 1-7 in inhibiting the growth and proliferation of bacteria.

9. The use of claim 8, wherein the compound traditional Chinese medicine bacteriostatic composition has the effect of inhibiting the growth and proliferation of escherichia coli, staphylococcus aureus, pseudomonas aeruginosa and candida albicans.