阅读说明：本技术 一种植物抑菌组合物及其应用 (Plant bacteriostatic composition and application thereof ) 是由 琚建伟 张守兵 于 2021-01-08 设计创作，主要内容包括：本发明公开了一种植物抑菌组合物及其应用,属于抗菌技术领域,所述植物抑菌组合物主要由以下植物提取物组成：藏红花提取物、厚朴提取物、红豆杉提取物和胡萝卜籽提取物。藏红花提取物具有抑制单核细胞增生李斯特菌和肠炎沙门氏菌的生长的作用；厚朴提取物具有抑制金黄色葡萄球菌生长的作用；红豆杉提取物具有具有抗炎、抗氧化、抗过敏、抗菌、抗病毒等作用；胡萝卜籽提取物具有抑制真菌的作用；植物抑菌组合物,具有良好的抑细菌和真菌的作用,同时具有温和安全不刺激的特点,可用于护肤品的绿色防腐剂。(The invention discloses a plant bacteriostatic composition and application thereof, belonging to the technical field of antibiosis, wherein the plant bacteriostatic composition mainly comprises the following plant extracts: saffron extract, magnolia bark extract, taxus chinensis extract and carrot seed extract. The stigma croci Sativi extract has effects of inhibiting growth of Listeria monocytogenes and Salmonella enteritidis; cortex Magnolia officinalis extract has effect in inhibiting the growth of Staphylococcus aureus; the Taxus chinensis extract has antiinflammatory, antioxidant, antiallergic, antibacterial, and antiviral effects; the carrot seed extract has antifungal effect; the plant bacteriostatic composition has good effects of inhibiting bacteria and fungi, has the characteristics of mildness, safety and no irritation, and can be used as a green preservative of skin care products.)

1. A plant bacteriostatic composition is characterized by mainly comprising the following plant extracts: saffron extract, magnolia bark extract, taxus chinensis extract and carrot seed extract.

2. The plant bacteriostatic composition according to claim 1, wherein the mass ratio of the saffron extract, the magnolia bark extract, the taxus chinensis extract and the carrot seed extract is 3-5:2-4:0.5-2: 1-3.

3. The plant bacteriostatic composition according to claim 1, wherein the extract site of saffron extract is flower, the extract site of magnolia bark extract is bark, the extract site of taxus chinensis extract is leaf, and the extract site of carrot seed extract is seed.

4. The plant bacteriostatic composition according to claim 1, wherein the preparation method of the plant extract comprises the following steps:

pulverizing the plant extract to obtain powder;

adding water and ethanol into the powder, uniformly stirring and ultrasonically oscillating to obtain a mixture;

refluxing the mixture under reduced pressure, and filtering to obtain extractive solution;

concentrating and drying the extracting solution to obtain plant extract solid;

dissolving with methyl propylene glycol and water, standing at low temperature, filtering, and collecting filtrate to obtain plant extract.

5. The plant bacteriostatic composition according to claim 1, wherein the detection method of the plant extract comprises the following steps:

detecting stigma croci Sativi extract with crocin-I as calibration;

detecting Magnolia officinalis extract with magnolol as calibration;

detecting the taxus chinensis extract by taking quercetin as a calibration substance;

the saffron extract was tested with carotol as the calibrator.

6. Use of a plant bacteriostatic composition according to any one of claims 1 to 5, wherein the plant bacteriostatic composition is used as a preservative for skin care products.

7. Use according to claim 6, wherein the phytosanitary composition is used to inhibit the growth and reproduction of one or a combination of the following microorganisms: staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger and Candida albicans.

8. The use of claim 6, wherein the skin care product comprises 0.8-5.0% by mass of the plant bacteriostatic composition.

9. The use of claim 8, wherein the skin care product further comprises 0.01-0.2 wt% of a chelating agent, 0.50-10.00 wt% of a polyhydric alcohol, 0.05-1.0 wt% of a thickener, 0.50-5.0 wt% of an emulsifier, 5.0-35.0 wt% of oil, 0.1-10.0 wt% of a skin conditioner, and 39.00-88.00 wt% of deionized water.

10. The use according to claim 9, wherein the skin care product is prepared by a process comprising:

adding a chelating agent, polyhydric alcohol and a thickening agent into deionized water to obtain a water phase;

fully and uniformly stirring the water phase and heating to 75-90 ℃;

mixing an emulsifier and grease to obtain an oil phase;

fully stirring the oil phase and heating to 75-90 ℃;

mixing the heated oil phase and the heated water phase, keeping the temperature and stirring for 20-30 minutes, and cooling;

adding skin conditioner and plant bacteriostatic composition during cooling to 40-50 deg.C;

and continuously stirring and cooling to obtain the skin care product.

Technical Field

The invention relates to the technical field of antibiosis, and particularly relates to a plant bacteriostatic composition and application thereof.

Background

The skin care product is very easily polluted by microorganisms in the natural environment and human bodies, in order to prevent the property and performance of the product from changing, a bacteriostatic agent is usually required to be added to prevent the product from deteriorating due to the proliferation of the microorganisms, the substance for inhibiting the growth of the microorganisms is usually called as a preservative, the preservative is usually a bacteriostatic agent but not a bactericide, the preservative has a relatively mild action mode and mainly plays the aim of inhibiting the growth and the proliferation of the microorganisms to gradually die, and the bactericide is usually easy to cause skin irritation and sensitivity due to excessively strong penetrating power to cell membranes and is rarely used for the purpose of preventing corrosion. The skin care product can be used with dozens of preservatives, mainly including formaldehyde-releasing agents, parabens, isothiazolinones, phenols, organic acids, halides, and the like. In recent years, the concept of 'no preservation' is created, and some products with bacteriostatic function outside the list of quasi-preservatives for cosmetics are produced, mainly comprising diol with high relative molecular mass, polyol monoester with high purity, organic metal ion chelating agent and the like, and the novel bacteriostatic agents greatly enrich bacteriostatic components in skin care products.

At present, although the commonly used preservatives are safe and effective within the limited range, consumers still have various misunderstandings on the addition of the preservatives, and the emerging bacteriostatic agent with the concept of 'no preservative' is usually derived from chemical synthesis and has some use limitations, such as the defects that glycols with high relative molecular mass are often smelled, have large addition amount, and are hot when being applied to the skin.

Disclosure of Invention

Aiming at the technical problems in the prior art, the invention provides a plant bacteriostatic composition and application thereof, wherein the plant bacteriostatic composition has a good bacteriostatic effect, is mild, safe and non-irritant, and is a green preservative for skin care products.

The invention discloses a plant bacteriostatic composition, which mainly comprises the following plant extracts: saffron extract, magnolia bark extract, taxus chinensis extract and carrot seed extract.

Preferably, the mass ratio of the saffron extract to the magnolia bark extract to the taxus chinensis extract to the carrot seed extract is 3-5:2-4:0.5-2: 1-3.

Preferably, the mass ratio of the saffron extract to the magnolia bark extract to the taxus chinensis extract to the carrot seed extract is 4:3:1: 2.

Preferably, the extract part of the saffron extract is flowers, the extract part of the magnolia bark extract is peels, the extract part of the taxus chinensis extract is leaves, and the extract part of the carrot seed extract is seeds.

Preferably, the preparation method of the plant extract comprises:

pulverizing the plant extract to obtain powder;

adding water and ethanol into the powder, uniformly stirring and ultrasonically oscillating to obtain a mixture;

refluxing the mixture under reduced pressure, and filtering to obtain extractive solution;

concentrating and drying the extracting solution to obtain plant extract solid;

dissolving with methyl propylene glycol and water, standing at low temperature, and filtering to obtain plant extract.

Preferably, the method for detecting the plant extract comprises the following steps: detecting stigma croci Sativi extract with crocin-I as calibration; detecting Magnolia officinalis extract with magnolol as calibration; detecting the taxus chinensis extract by taking quercetin as a calibration substance; the saffron extract was tested with carotol as the calibrator.

Preferably, the plant bacteriostatic composition is applied to a preservative of a skin care product.

Preferably, the plant bacteriostatic composition is used for inhibiting the growth and reproduction of one or a combination of the following microorganisms: staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger and Candida albicans.

Preferably, the skin care product contains 0.8-5.0% of plant bacteriostatic composition by mass percent.

Preferably, the skin care product also comprises 0.01-0.2 wt% of chelating agent, 0.50-10.00 wt% of polyalcohol, 0.05-1.0 wt% of thickening agent, 0.50-5.0 wt% of emulsifier, 5.0-35.0 wt% of grease, 0.1-10.0 wt% of skin conditioner and 39.00-88.00 wt% of deionized water.

Preferably, the preparation method of the skin care product comprises the following steps: adding a chelating agent, polyhydric alcohol and a thickening agent into deionized water to obtain a water phase; fully and uniformly stirring the water phase and heating to 75-90 ℃; mixing an emulsifier and grease to obtain an oil phase; fully stirring the oil phase and heating to 75-90 ℃; mixing the heated oil phase and the heated water phase, keeping the temperature and stirring for 20-30 minutes, and cooling; adding skin conditioner and plant bacteriostatic composition during cooling to 40-50 deg.C; and continuously stirring and cooling to obtain the skin care product.

Compared with the prior art, the invention has the beneficial effects that: the stigma croci Sativi extract has effects of inhibiting growth of Listeria monocytogenes and Salmonella enteritidis; cortex Magnolia officinalis extract has effect in inhibiting the growth of Staphylococcus aureus; the Taxus chinensis extract has antiinflammatory, antioxidant, antiallergic, antibacterial, and antiviral effects; the carrot seed extract has antifungal effect; the plant bacteriostatic composition has good effects of inhibiting bacteria and fungi, has the characteristics of mildness, safety and no irritation, and can be used as a green preservative of skin care products.

Drawings

FIG. 1 is a flow chart of a method for preparing a plant extract;

FIG. 2 is a HPLC peak chart of crocin-I;

figure 3 is an HPLC peak profile of magnolol.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.

The invention is described in further detail below with reference to the attached drawing figures:

a plant bacteriostatic composition mainly comprises the following plant extracts: saffron extract, magnolia bark extract, taxus chinensis extract and carrot seed extract.

Wherein the saffron extract has the effect of inhibiting the growth of Listeria monocytogenes (Listeria monocytogenes) and Salmonella enteritidis (Salmonella enterica); cortex Magnolia officinalis extract has effect in inhibiting the growth of Staphylococcus aureus; the Taxus chinensis extract has antiinflammatory, antioxidant, antiallergic, antibacterial, and antiviral effects; the carrot seed extract has antifungal effect; experiments prove that the plant bacteriostatic composition has good functions of inhibiting bacteria and fungi, has the characteristics of being mild, safe and non-irritating, and can be used as a green preservative of a skin care product.

In the specific implementation, the extraction part of the saffron extract is flowers, the extraction part of the magnolia officinalis extract is peels, the extraction part of the taxus chinensis extract is leaves, and the extraction part of the carrot seed extract is seeds.

As shown in fig. 1, the preparation method of the plant extract comprises:

step 101: pulverizing the plant extract to obtain powder. The powder can be pulverized to more than 100 mesh.

Step 102: adding water and ethanol into the powder, stirring uniformly and carrying out ultrasonic oscillation to obtain a mixture. The present invention extracts the effective components of the plant extract part by the mixed solution of water and ethanol, but is not limited thereto.

Step 103: refluxing the mixture at 70-80 deg.C under reduced pressure for 6-10 hr, and filtering to obtain extractive solution.

Step 104: concentrating and drying the extract to obtain plant extract solid. Concentrated and dried to remove the solvent.

Step 105: dissolving plant extract solid with methyl propylene glycol and deionized water, standing at 2-8 deg.C, filtering, and collecting filtrate to obtain plant extract. The filtrate was used as a plant extract.

The preparation method of the plant extract is suitable for saffron extract, magnolia bark extract, taxus chinensis extract and carrot seed extract.

The detection method of the plant extract comprises the following steps: detecting stigma croci Sativi extract with crocin-I as calibration material, in a specific embodiment, its content threshold is more than or equal to 0.5%; detecting cortex Magnolia officinalis extract with magnolol as calibration substance, wherein the content threshold is not less than 0.5%; detecting the extract of the taxus chinensis by using quercetin as a calibration substance, wherein the content threshold value of the extract is more than or equal to 0.1 percent; the content threshold value of the saffron extract is more than or equal to 0.1 percent by using the daucosterol as a calibration substance for detecting the saffron extract. The extraction quality of the plant extract is managed by detecting the plant extract through one main component, a content threshold value can be set according to the extraction process and the condition of an extraction part, and the content threshold value is used for judging the extraction quality of the plant extract.

Wherein the crocin-I, magnolol, quercetin and carotene can be detected by High Performance Liquid Chromatography (HPLC). For example: FIG. 2 is a HPLC peak pattern of crocin-I, the peak time of crocin-I is 18.5 minutes; figure 3 is an HPLC peak profile of magnolol with a peak time of 9.2 minutes. The content of the calibration material can be detected by the proportion of the peak area of the calibration material; quercetin and carotenol can be detected by the same or similar HPLC method.

The mass ratio of the saffron extract to the magnolia bark extract to the taxus chinensis extract to the carrot seed extract is 3-5:2-4:0.5-2:1-3, preferably 4:3:1: 2.

Example 1

Mixing 3g stigma croci Sativi extract, 2g cortex Magnolia officinalis extract, 2g Taxus chinensis extract, and 1g radix Dauci Sativae seed extract to obtain composition A.

Example 2

And uniformly mixing 4g of saffron extract, 3g of mangnolia officinalis extract, 1g of taxus chinensis extract and 2g of carrot seed extract to obtain the composition B.

Example 3

Mixing 5g stigma croci Sativi extract, 4g cortex Magnolia officinalis extract, 0.5g Taxus chinensis extract, and 3g radix Dauci Sativae seed extract to obtain composition C.

Example 4

Mixing 3.5g stigma croci Sativi extract, 3g cortex Magnolia officinalis extract, 1.5g Taxus chinensis extract, and 2.5g radix Dauci Sativae seed extract to obtain composition D.

Staphylococcus aureus inhibition of the compositions obtained in examples 1 to 4, respectivelyPreparing an experiment: 9.6*10⁶cfu/g of Staphylococcus aureus suspension to which 1% of the composition was added. Wherein the composition B has high 14-day rejection rate, and the number of detected bacteria is less than 10 cfu/g.

Challenge test:

taking the composition B to carry out a 28-day microbial challenge test, wherein the test method is a method for evaluating the preservative effect by referring to the microbial challenge test on the United states Pharmacopeia: after the bacteria are cultured for 14 days, the number of the bacteria is reduced to be below 0.1 percent of the original number, and the number of the bacteria is not increased to pass the test in the subsequent test; the fungus remained below the original level after 14 days of culture and no further increase in subsequent tests was a pass test.

The specific scoring criteria were:

initial inoculum size of bacteria 10⁶cfu/g～10⁷cfu/g, mold 10⁵cfu/g～10⁶cfu/g; (28 th day), the sample contains bacteria or fungi > 10³cfu/g, the sample can not pass a corrosion prevention and bacteriostasis challenge test, which shows that the sample can not effectively play a role in inhibiting the growth of microorganisms, and the product is easily polluted by the microorganisms during production, storage and use; ② on day 28, the sample contains bacteria or mold 10²cfu/g～10³cfu/g, the sample conditionally passes a challenge test, namely when the protein or other animal and plant material components in the product are not particularly high, the produced sanitary environment meets the requirement, and the packaging material is not easy to generate secondary pollution, the bacteriostatic combination meets the requirement and can be used, otherwise, the bacteriostatic combination cannot be used; and thirdly, on the 28 th day, bacteria or mould contained in the sample cannot be detected, which shows that the sample has outstanding antiseptic and bacteriostatic ability and has stronger inhibition effect on microorganisms, and the product is not easily polluted by the microorganisms during production, storage and use through challenge tests.

The specific detection method comprises the following steps:

respectively culturing Staphylococcus aureus (Staphylococcus aureus, ATCC 6538), escherichia coli (ATCC 8739) and pseudomonas aeruginosa (pseudomonas aeruginosa, ATCC 9027) at 36 +/-1 ℃ for 36-48 hours;

respectively picking appropriate amount of bacterial colony, mixing with sterile normal saline, and making into final productMixing the bacterial suspensions to a total concentration of about 1.0X 10⁸cfu/ml；

Aspergillus niger (ATCC 16404) at 27 + -1 deg.C for 120 hours, Candida albicans (ATCC 10231) at 27 + -1 deg.C for 24 hours;

respectively picking appropriate amount of Aspergillus niger and Candida albicans colonies, and mixing with sterile normal saline to obtain mixed fungus suspension with total concentration of 1.0 × 10⁷cfu/ml；

Taking skin care products, setting 6 groups of samples in total, wherein the adding amounts of the composition B are respectively 0.3%, 0.5%, 0.8%, 1.2%, 1.5% and 2.0%, and 6 samples with concentration gradients are formed; the samples are added respectively to the solution with the final bacterial content of 1.0 multiplied by 10⁶～5.0×10⁶CFU/g mixed bacterial suspension, final bacteria content 1.0X 10⁵～5.0×10⁵CFU/g mixed fungus suspension, and fully mixing uniformly, and culturing 6 groups of samples at 36 +/-1 ℃;

samples were taken on day 0, day 3, day 7, day 14, day 21 and day 28 of inoculation for bacterial load analysis. The test data are shown in the following table:

as shown in the above table:

(1) on day 28, the number of bacteria and fungi in each sample was less than 10³cfu/g, which shows that different concentrations of the plant bacteriostatic composition have different degrees of killing and inhibiting effects on microorganisms, and 3#, 4#, 5# and 6# samples pass challenge tests according to the rating standard, so that the plant bacteriostatic composition is not easily polluted by microorganisms during production, storage and use;

(2)3#, 4#, 5# and 6# samples, with the increase of the concentration of the plant bacteriostatic composition, the total number of bacterial colonies is not detected (less than 10cfu/g) at 14 days, the fungus only contains 3# samples with lower data volume, which shows that the bacteriostatic effect is obvious after 14 days, the good effect of inhibiting microorganisms can be achieved when the concentration of the composition B exceeds 0.8%, and the product is not easily polluted by microorganisms during production, storage and use;

(3)4#, 5#, 6# sample, from 14 days on, no bacterium and mould were detected, showed excellent bacteriostatic and preservative functions.

Thus, the above experiments demonstrate that: the 3#, 4#, 5# and 6# samples pass the challenge tests, the higher the addition amount of the plant bacteriostatic composition is, the better the preservative effect is, the concentration of the composition B is more than 0.8%, the preservative effect is excellent, and the product is not easily polluted by microorganisms during production, storage and use. The plant bacteriostatic composition of the invention can be used for inhibiting the growth and reproduction of the following microorganisms or the combination thereof: staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Aspergillus niger and Candida albicans.

The skin care product contains: 0.8 to 2.0 percent of plant bacteriostatic composition, 0.01 to 0.2 percent of chelating agent, 0.50 to 10.00 percent of polyol, 0.05 to 1.0 percent of thickening agent, 0.50 to 5.0 percent of emulsifier, 5.0 to 35.0 percent of grease, 0.1 to 10.0 percent of skin conditioner and 39.00 to 88.00 percent of deionized water.

The preparation method of the skin care product comprises the following steps:

adding a chelating agent, polyhydric alcohol and a thickening agent into deionized water to obtain a water phase;

fully and uniformly stirring the water phase and heating to 75-90 ℃;

mixing an emulsifier and grease to obtain an oil phase;

fully stirring the oil phase and heating to 75-90 ℃;

mixing the heated oil phase and the heated water phase, keeping the temperature and stirring for 20-30 minutes, and cooling;

adding skin conditioner and plant bacteriostatic composition during cooling to 40-50 deg.C;

and continuously stirring and cooling to obtain the skin care product.

However, the application range of the plant bacteriostatic composition is not limited to the above range, and the plant bacteriostatic composition can also be used for other types of skin care products.

Safety test

The safety test, the cosmetic eye irritation/corrosiveness test for chick chorioallantoic membrane (HET-CAM test), was performed using the industry standard SN/T2329-2009.

Dissolving the composition B in normal saline to prepare a test sample with the weight of 5 percent, wherein chicken embryos adopt a Lehang strain, the age of the chicken embryos is 9 days, and the test conditions are as follows: 37.5 ℃, relative humidity: 50% -70%, action time: 180s, and simultaneously using physiological saline as a negative control and 1% SLS as a positive control, according to an end point evaluation method, the detection result is ES-6, and the result is evaluated: no/light irritability. I.e. composition B is mild, safe and non-irritating at use levels below 5% wt. Therefore, the plant bacteriostatic composition with the mass percent of 0.8-5.0% is a reasonable use dose.

The above is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.