CN107224408B

CN107224408B - Plant-derived broad-spectrum antibacterial agent and preparation method and application thereof

Info

Publication number: CN107224408B
Application number: CN201610169716.3A
Authority: CN
Inventors: 翟春涛; 贺改英; 蔡艳; 崔水兴
Original assignee: Laibo Cosmetics Technology Shanghai Co ltd; Shanghai Shengwei Biological Technology Co ltd
Current assignee: LAIBO COSMETICS TECHNOLOGY (SHANGHAI) Co.,Ltd.; Shanghai Zhina Biotechnology Co.,Ltd.
Priority date: 2016-03-23
Filing date: 2016-03-23
Publication date: 2020-06-02
Anticipated expiration: 2036-03-23
Also published as: CN107224408A

Abstract

The invention discloses a botanical broad-spectrum antibacterial agent and a preparation method and application thereof, wherein the raw materials comprise a material A, water, lipase and a pH regulator, and the material A comprises, by mass, 100 parts of litsea cubeba oil, 0.1-5 parts of patchouli oil, 0.1-5 parts of citronella oil, 0.1-5 parts of hydrolyzed ginsenoside and 80-99.6 parts of caprylic/capric triglyceride; the preparation method comprises the following steps: 1) uniformly mixing all the components in the material A to obtain a grease mixture; 2) uniformly mixing water and lipase, adjusting the pH value to 7-10, uniformly mixing the mixture with the grease mixture, and stirring for enzymolysis at 40-50 ℃; 3) performing ultrasonic treatment on the enzymolysis liquid, then performing oil-water separation, and taking an oil phase; 4) aging the oil phase, and filtering the supernatant to obtain filtrate. The plant-derived broad-spectrum antibacterial agent provided by the invention has the advantages of broad-spectrum antibacterial, mildness, good solubility, economy, small dosage, stability and suitability for industrial production.

Description

Plant-derived broad-spectrum antibacterial agent and preparation method and application thereof

Technical Field

The invention relates to a botanical broad-spectrum antibacterial agent, a preparation method and application thereof.

Background

Preservatives are substances which prevent the growth of microorganisms. In cosmetics, preservatives serve to protect the product from microbial contamination. Most of the preservatives commonly used at present are synthetic preservatives, such as hydantoin (DMDMH), cason series, bronopol, iodopropynyl butyl carbamate (IPBC), parabens, triclosan, and the like. These preservatives all have certain drawbacks: for example, DMDMH is easy to release methanol in the using process, and has potential safety hazard; the cason series may generate certain irritation to certain skin types in the use process due to the chlorine-containing component; bronopol is prone to produce carcinogenic nitrosamines; IPBC may lead to excessive iodine intake by the user; the parabens are easy to accumulate in adipose tissues; triclosan has been shown to have negative environmental effects; furthermore, the paraben preservatives have been shown in specific test systems to mimic the ability of the female hormone estrogen, with negative effects on human safety and physiological systems. Therefore, the market needs to present a natural preservative with low toxicity, high efficiency and no pollution.

In recent years, under the hot tide impact of returning to nature and enjoying green and health, the addition of purely natural preservatives into cosmetics becomes a hot spot advocated and developed at home and abroad, and the extraction and separation of natural preservatives which have strong antibacterial property, broad range, safety and no toxicity from natural products becomes an important direction for future development.

Most of the natural preservatives used at present are crude products, which have darker color, stronger smell and narrower antibacterial spectrum. Meanwhile, some traditional Chinese medicines are not as good as single medicines in antibacterial effect after combined use, for example, the antibacterial effect of the mixture of astragalus and scutellaria on escherichia coli, pseudomonas aeruginosa and salmonella is weaker than that of the single medicine after the mixture is mixed and matched. Therefore, the development of an antibacterial agent with natural raw materials and broad-spectrum antibacterial property is a technical problem to be solved in the field.

Disclosure of Invention

The invention aims to overcome the defect of narrow antibacterial spectrum of a natural preservative in the prior art, and provides a botanical broad-spectrum antibacterial agent and a preparation method and application thereof. The plant-derived broad-spectrum antibacterial agent provided by the invention has the advantages of broad-spectrum antibacterial property, mildness, good solubility, economy, small dosage, stability and suitability for industrial production, can effectively inhibit the growth of microorganisms in cosmetics, prolongs the shelf life of the cosmetics, is beneficial to ensuring the safety of products, and prevents possible infection caused by using products polluted by microorganisms by consumers. The preparation method provided by the invention overcomes the problems of heavy smell, less retention of effective components, more impurities, narrow antibacterial spectrum, large dosage, strong irritation and the like of the traditional Chinese medicine extract prepared by the traditional preparation process.

The invention adopts the following technical scheme to solve the technical problems:

the invention provides a preparation method of a botanical broad-spectrum antibacterial agent, which comprises the following raw materials of a material A, water, lipase and a pH regulator, wherein the material A comprises the following components in parts by mass: 0.1-5 parts of litsea cubeba oil, 0.1-5 parts of patchouli oil, 0.1-5 parts of citronella oil, 0.1-5 parts of hydrolyzed ginsenoside and 80-99.6 parts of caprylic capric triglyceride; the preparation method comprises the following steps:

(1) uniformly mixing all the components in the material A to obtain a grease mixture;

(2) uniformly mixing water and lipase, adjusting the pH value to 7-10, uniformly mixing the mixture with the grease mixture, performing enzymolysis at 40-50 ℃, and stirring in the enzymolysis process to obtain an enzymolysis liquid;

(3) carrying out ultrasonic treatment on the enzymolysis liquid, then carrying out oil-water separation, and taking an oil phase;

(4) and (3) aging the oil phase, and filtering the supernatant to obtain a filtrate.

In the invention, the material A preferably comprises the following components in parts by mass as 100 parts: 0.2-1 part of litsea cubeba oil, 0.2-1 part of patchouli oil, 0.2-1 part of citronella oil, 0.2-1 part of hydrolyzed ginsenoside and 96-99.2 parts of caprylic capric triglyceride.

Wherein said litsea cubeba oil has the meaning conventional in the art, is derived from the fruit of the plant Lauraceae, Zanthoxylum piperitum, preferably from the Gerano optical perfumery.

Wherein said patchouli oil has the meaning conventional in the art and is derived from aerial parts of the labiatae plant patchouli, preferably available from trillion perfume oil limited, ge' an.

Wherein said citronella oil has the meaning conventionally used in the art, and is derived from the whole grass of the citronella plant citronella grass of the family Gramineae, preferably from the trillion perfume oil Co.

Wherein the hydrolyzed ginsenosides have a meaning conventionally used in the art, and are derived from a hydrolysate of ginsenosides in the leaves of roots and stems of Panax ginseng C.A. Meyer, Araliaceae, preferably from Cissampel extract Biotech Co.

Wherein the caprylic capric triglyceride has the conventional meaning in the field, is derived from the fruit of oil palm of the family Palmae, is a refined derivative of palm oil, and is preferably purchased from Shanghai Saifu chemical development Co.

In the invention, the water is a conventional substance in the field, preferably deionized water, and the dosage of the deionized water is the conventional dosage in the field, preferably 1/10-1/5 of the dosage of the material A.

In the present invention, the pH regulator is a substance conventional in the art, and preferably one or more of sodium hydroxide, sodium carbonate, sodium bicarbonate, hydrochloric acid, and citric acid.

In step (2), the lipase is a substance conventional in the art, preferably purchased from banksia jenno bio-enzyme limited; the dosage of the lipase is the enzyme activity required for adjusting the enzyme activity in the solution to the conventional enzymolysis reaction, preferably the dosage of the lipase required for adjusting the enzyme activity to 10-500 u/g, and more preferably the dosage of the lipase required for adjusting the enzyme activity to 50-100 u/g.

In the step (2), the enzymolysis time is the conventional enzymolysis time in the field, and is generally more than 1 hour, preferably 1 to 4 hours.

In the step (2), the stirring is performed by a conventional method in the art, and the stirring speed is preferably 100 to 300rpm, more preferably 200 rpm.

In the step (3), the ultrasound is performed by a conventional method in the field, and the frequency of the ultrasound is preferably 25000 to 35000Hz, and more preferably 30000 Hz; the time of the ultrasonic treatment is preferably 2 to 10 minutes.

In the step (3), the oil-water separation can be realized by adopting conventional operations in the field, and preferably, standing is adopted. The standing time is conventional in the field, and is preferably 0.5-4 h.

In the step (4), the aging is performed by the conventional operation in the field, and the aging temperature is preferably-6 ℃ to-2 ℃, and more preferably-4 ℃; the aging time is preferably 8 to 14 hours, and more preferably 12 hours.

The invention also provides the plant-derived broad-spectrum antibacterial agent prepared by the preparation method.

In the present invention, the plant-derived broad-spectrum antibacterial agent has broad-spectrum antibacterial performance, and the species of bacteria that can be resisted preferably include one or more of staphylococcus aureus, escherichia coli, pseudomonas aeruginosa, candida albicans and aspergillus niger.

The invention also provides application of the plant-derived broad-spectrum antibacterial agent as a preservative in daily cosmetics and/or skin external medicines.

On the basis of the common knowledge in the field, the above preferred conditions can be combined randomly to obtain the preferred embodiments of the invention.

The reagents and starting materials used in the present invention are commercially available.

The positive progress effects of the invention are as follows:

(1) the invention leads the extract to have broad-spectrum antibacterial action by applying the compatibility and the synergistic action among plants, and introduces the hydrolyzed ginsenoside into the raw materials, thus greatly improving the safety of the antibacterial agent;

(2) the preparation method integrates the modern biological enzymolysis technology in the preparation process of the traditional Chinese medicine preparation, obviously reduces the irritation, greatly improves the antibacterial activity of the plant antibacterial agent, and improves the influence of bad smell on the application of the antibacterial agent; part of impurities are removed through aging, so that small molecular substances are better reserved, and the cosmetic has important contribution to meeting the use requirement and aesthetic feeling of cosmetics;

(3) the whole process of the invention takes vegetable oil and extract as raw materials, and the preparation is carried out under the condition of low temperature, thus being green and natural, energy-saving and environment-friendly;

(4) the antibacterial agent prepared by the invention can be mutually dissolved with the dipropylene glycol in any proportion, and can be diluted by the dipropylene glycol for convenient weighing due to low use amount.

Detailed Description

The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions.

In the following examples, the litsea cubeba oil used was purchased from the Jian national photo perfumery, patchouli oil was purchased from the trillion perfume oil Co.Ltd, Geranium oil was purchased from the trillion perfume oil Co.Ltd, hydrolyzed ginsenosides were purchased from the Sian extractive Biotech Co.Ltd, caprylic capric triglyceride was purchased from the Shanghai Saifu chemical development Co.Ltd, and lipase was purchased from the Jiuno bioenzyme Co.Ltd, Zaozhuang.

Example 1 (antibacterial agent)

The preparation method of the plant-derived broad-spectrum antibacterial agent in the embodiment is as follows:

(1) mixing 0.1g Litsea cubeba oil, 0.1g patchouli oil, 0.1g citronella oil, 0.1g hydrolyzed ginsenoside and 99.6g caprylic capric triglyceride to obtain oil mixture;

(2) adding 10g of deionized water into lipase until the enzyme activity is 50u/g, adjusting the pH value to 8 by using sodium bicarbonate, uniformly mixing the lipase and the grease mixture in the step (1), carrying out enzymolysis for 2h at 40 ℃, stirring in the enzymolysis process, wherein the stirring speed is 100 rpm;

(3) stopping stirring, performing ultrasonic treatment for 2 minutes under the condition of 30000Hz, standing for 30 minutes for oil-water separation, and taking an oil phase;

(4) and (3) aging the oil phase at-4 ℃ for 12 hours, taking the supernatant and filtering to obtain the plant-derived broad-spectrum antibacterial agent.

Example 2 (antibacterial agent)

(1) mixing 1g of pungent litse fruit oil, 1g of patchouli oil, 1g of citronella oil, 1g of hydrolyzed ginsenoside and 96g of caprylic capric triglyceride to obtain a grease mixture;

(2) adding 10g of deionized water into lipase until the enzyme activity is 100u/g, adjusting the pH value to 9 by using sodium carbonate, uniformly mixing the mixture with the grease mixture in the step (1), carrying out enzymolysis for 4h at 45 ℃, stirring in the enzymolysis process, wherein the stirring speed is 200 rpm;

(3) stopping stirring, performing ultrasonic treatment for 5 minutes under the condition of 30000Hz, standing for 1 hour for oil-water separation, and taking an oil phase;

Example 3 (antibacterial agent)

(1) fully and uniformly mixing 5g of litsea cubeba oil, 5g of patchouli oil, 5g of citronella oil, 5g of hydrolyzed ginsenoside and 80g of caprylic capric triglyceride to obtain a grease mixture;

(2) adding 20g of deionized water into lipase until the enzyme activity is 10u/g, adjusting the pH value to 7 by using citric acid, uniformly mixing the lipase and the grease mixture in the step (1), carrying out enzymolysis for 4h at 50 ℃, stirring in the enzymolysis process, wherein the stirring speed is 300 rpm;

(3) stopping stirring, performing ultrasonic treatment for 10 minutes under 25000Hz, standing for 4 hours for oil-water separation, and taking an oil phase;

(4) and (3) aging the oil phase at-6 ℃ for 8 hours, taking the supernatant and filtering to obtain the plant-derived broad-spectrum antibacterial agent.

Example 4 (antibacterial agent)

(1) mixing 0.2g of pungent litse fruit oil, 0.2g of patchouli oil, 0.2g of citronella oil, 0.2g of hydrolyzed ginsenoside and 99.2g of caprylic/capric triglyceride to obtain a grease mixture;

(2) taking 10g of deionized water, adding lipase until the enzyme activity is 500u/g, adjusting the pH value to 10 by using sodium hydroxide, uniformly mixing with the grease mixture in the step (1), carrying out enzymolysis for 4h at 45 ℃, stirring during the enzymolysis process, wherein the stirring speed is 200 rpm;

(3) stopping stirring, performing ultrasonic treatment at 35000Hz for 5min, standing for 1 hr for oil-water separation, and collecting oil phase;

(4) aging the oil phase at-2 deg.C for 14 hr, collecting supernatant, and filtering to obtain the plant-derived broad-spectrum antibacterial agent.

Comparative example 1 (comparative example of antibacterial agent without biological enzymolysis technique)

The preparation of the antibacterial agent in this comparative example was as follows:

(1) mixing 1g of pungent litse fruit oil, 1g of patchouli oil, 1g of citronella oil, 1g of hydrolyzed ginsenoside and 96g of caprylic capric triglyceride;

(2) aging at-10 deg.C for 12 hr, and filtering to obtain the plant-derived antiseptic.

Comparative example 2 (comparative example of antibacterial agent without hydrolysis of ginsenoside in raw Material)

(1) mixing 1g of pungent litse fruit oil, 1g of patchouli oil, 1g of citronella oil and 97g of caprylic capric triglyceride;

(2) adding 10g of deionized water into lipase until the enzyme activity is 100u/g, adjusting the pH value to 9, mixing with the grease in the step (1), and stirring for 4 hours at the temperature of 45 ℃;

(3) stopping stirring, performing ultrasonic treatment for 5 minutes at 30000Hz, standing for 1h, performing oil-water separation, and collecting oil phase;

(4) aging at-10 deg.C for 12 hr, and filtering to obtain the plant-derived antiseptic.

Effect example 1 (antibacterial test of antibacterial agent)

The bacteriostatic test comprises the following steps:

(1) test strains

Staphylococcus aureus (ATCC 8739), Escherichia coli (ATCC 6538), Pseudomonas aeruginosa (ATCC9027), Candida albicans (ATCC 10231), Aspergillus niger (ATCC 16404).

(2) Culture medium

TSB solid medium: peptone 10g/L, NaCl 10g/L, K₂HPO₄2.5g/L, yeast extract 3g/L, agar 15g/L, dissolved in distilled water, and adjusted pH to 7.2Performing autoclaving at 115 ℃ for 20min +/-0.2; TSB liquid medium: peptone 10g/L, NaCl 10g/L, K₂HPO₄2.5g/L yeast extract 3g/L, dissolving in distilled water, adjusting pH to 7.2 + -0.2, and autoclaving at 115 deg.C for 20 min;

SDB solid medium: peptone 10g/L, glucose 20g/L, K₂HPO₄3g/L, 5g/L yeast extract, 15g/L agar, dissolving with distilled water, and autoclaving at 115 deg.C for 20 min; SDB liquid medium: peptone 10g/L, glucose 20g/L, K₂HPO₄3g/L, 5g/L yeast extract, dissolving in distilled water, and autoclaving at 115 deg.C for 20 min.

(3) Bacterial suspension preparation and inoculation

Coating 100 μ L of glycerol-preserved Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa on TSB solid culture medium, culturing at 37 deg.C for 2d, resuspending with normal saline to obtain bacterial suspension, counting by plate culture method, and adjusting the concentration of test bacteria to 10⁵～10⁶CFU/mL。

Spreading Candida albicans and Aspergillus niger 100 μ L on SDB solid culture medium, culturing at 30 deg.C for 2d, re-suspending with normal saline to obtain bacterial suspension, counting by plate culture method, and adjusting test bacteria concentration to 10⁵～10⁶。

(4) Determination of sample bacteriostasis

Cup and dish method

TSB or SDB solid medium was poured into the petri dish, and after solidification, 100. mu.L of test bacteria was spread evenly on the medium. 2 sterile Oxford cups (inner diameter (6.0 +/-0.1) mm, outer diameter (8.0 +/-0.1) mm and height (10.0 +/-0.1) mm are equidistantly placed on a culture medium, 200 mu L of samples to be tested are respectively injected into the cups, and physiological saline or physiological saline containing a certain amount of DMSO is used as a control. After culturing in a thermostat at 37 ℃ or 30 ℃ for 2 days, the diameters of the inhibition zones of various medicines are measured by a ruler, and the average value is taken.

Microplate method

Adding 100 μ L of sample to be tested (initial concentration), adding 1.0 × 10 concentration of test bacteria⁵CFU/mL is 2 XTSB or 2 XSDB liquid culture medium 100 mu L, and the mixture is mixed evenly, namely the test concentration of the sample to be tested is initial50 wt% of the concentration. After culturing in an incubator at 30 ℃ or 37 ℃ for 2d, aseptic growth is observed. The premise of result judgment is that the growth control is good, and the blank control aseptically grows clearly.

(5) Determination of minimum inhibitory concentration (MIC value)

① the determination method of the concentration of the lowest inhibitory staphylococcus aureus (MIC), the concentration of the lowest inhibitory escherichia coli (MIC), the concentration of the lowest inhibitory pseudomonas aeruginosa (MIC) and the concentration of the lowest inhibitory candida albicans (MIC) is as follows:

preparing a liquid medicine: the sample to be tested is diluted with normal saline to the experimental concentration.

Preparing bacterial liquid: the bacterial suspension was diluted with 2 XSSB or 2 XSSB liquid medium to a final bacterial concentration of l0⁵CFU/mL。

Culturing and judging results: adding 100 μ L of bacteria solution and 100 μ L of liquid medicine into the micropores, setting negative control without bacteria and normal growth control without liquid medicine, making 3 parallels for each medicine, and taking average value. And (3) incubating the mixture in a wet box at 37 ℃ or 30 ℃, observing the result after 48 hours, and reading the data by adopting a direct method. The premise of result judgment is that the growth contrast is good, the blank contrast aseptically grows clearly, and the growth of bacteria in other holes is inhibited along with the increase of the drug concentration gradient.

measurement of Minimum Inhibitory Concentration (MIC) of Aspergillus niger

Pouring SDB solid culture medium into a culture dish by cup-dish method, and adding Aspergillus niger (10) after coagulation⁵CFU/ml) 100. mu.L, and was uniformly applied. 3 sterile Oxford cups (inner diameter (6.0 + -0.1) mm, outer diameter (8.0 + -0.1) mm, height (10.0 + -0.1) mm) were placed equidistantly on the medium, and 200. mu.L of samples (diluted with physiological saline) of different concentrations were injected into the cups, respectively. A normal growth control and a saline control were also set without the test sample. The premise of the judgment of the result is that the growth control and the normal saline control grow well, and the growth of other bacteria is inhibited along with the increase of the concentration gradient of the medicament.

Compared with single vegetable oil (vegetable oil is diluted into 3% solution by caprylic capric triglyceride) and known caprylic capric triglyceride with certain antiseptic effect possibly generated in the hydrolysis process, the bacteriostatic result of the product is shown in table 1:

TABLE 1 comparison of MIC values for examples with vegetable oils alone

As can be seen from Table 1, within 100. mu.L/mL, litsea cubeba oil has no antibacterial effect on Escherichia coli and Aspergillus niger, and has a general antibacterial effect on Candida albicans and Pseudomonas aeruginosa; the patchouli has no antibacterial effect on pseudomonas aeruginosa within 100 mu L/mL, and has common antibacterial effect on escherichia coli and staphylococcus aureus; within 100 mu L/mL, the citronella oil has no antibacterial effect on escherichia coli and pseudomonas aeruginosa, the antibacterial effect on staphylococcus aureus, candida albicans and aspergillus niger is general, the antibacterial effect on pseudomonas aeruginosa and aspergillus niger is general by glyceryl monocaprylate, and the antibacterial effect on pseudomonas aeruginosa and candida albicans is general by glyceryl monocaprate.

The bacteriostatic results of the inventive example and the comparative example are shown in table 2:

TABLE 2 MIC value comparison of examples and comparative examples

As is clear from Table 2, the inhibitory effect of comparative example 1 on each bacterium is general, and the inhibitory effect of comparative example 2 is equivalent to that of example 2.

It can be seen from this that: firstly, the reasonable compatibility of plants can achieve the broad-spectrum antibacterial effect; secondly, each bacterium is seen independently, the antibacterial effect after compatibility is superior to that of a single plant, the result that 1+1 is greater than 2 is achieved, and the synergistic effect on the antibacterial effect among the plants is reflected; thirdly, the comprehensive product after enzymolysis is superior to the possible bacteriostatic effect of a single component; and fourthly, the antibacterial effect can be greatly improved after enzymolysis.

The formula of the invention has obvious bacteriostatic effect, broad-spectrum antibacterial effect, better bacteriostatic effect than that of a single medicine, and overcomes the limitation of the single medicine to certain strains. Meanwhile, the proportion of the three traditional Chinese medicines can inhibit the growth of bacteria to the maximum extent. The product is suitable for being used as a plant preservative, the process scheme keeps the antibacterial effect, the color of the product is greatly weakened, the product is stable, and the product is suitable for being added into cosmetics to be used as a preservative.

To further illustrate the technical content, the objects and effects achieved by the present invention, the present invention will be further illustrated in the following examples, in which the applicant expressly adds collagen and glucose which are susceptible to the growth of microorganisms to effectively explain the effect of the plant preservative in the present process, and the examples are not intended to limit the present invention in any way. All materials were in mass percent concentration.

Example 5 (use of the antibacterial agent prepared in example 1 of the present invention as a preservative in a cream-type product)

The cream product of the embodiment can be used as moisturizing cream, repairing cream and the like, and the raw material formula is as follows:

the process for preparing the cream product of this example is as follows:

(1) adding glycerol into a water phase kettle, adding sodium hyaluronate and sodium polyacrylate, stirring at 300rpm, adding water and disodium ethylene diamine tetraacetate into the kettle, heating to 85 deg.C, and dissolving to clarify to obtain water phase.

(2) Placing caprylic acid silicic acid triglyceride, behenyl alcohol, VE acetic ester, emulgadeSurro (trade name is emulgadeSurro, specific substances are sucrose polystearate and hydrogenated polyisobutene) and phytosterol into an oil phase kettle, heating to 85 deg.C, and stirring at 300rpm to obtain oil phase.

(3) Mixing the oil phase (2) and the water phase (1) in an emulsifying kettle, homogenizing (speed 500rpm) for 5min, stirring and cooling to 45 ℃.

(4) Adding the antibacterial agent obtained in the embodiment 1, collagen, glucose and 1, 2-octanediol into the mixture (3), stirring at 500rpm, and cooling to room temperature.

Example 6 (use of the antibacterial agent prepared in example 2 of the present invention as a preservative in a cream-type product)

the process for preparing the cream product of this example is as follows:

(2) Placing caprylic acid triglyceride silicate, behenyl alcohol, VE acetate, Emulgade Sucro and phytosterol into an oil phase kettle, heating to 85 deg.C, and stirring at 300rpm to obtain oil phase.

(4) Adding the natural antibacterial agent obtained in the embodiment 2, collagen, glucose and 1, 2-octanediol into the mixture (3), stirring at 500rpm, and cooling to room temperature.

Example 7 (use of the antibacterial agent prepared in example 1 of the present invention as a preservative in aqueous products)

The water product of the embodiment can be used as skin lotion, moisturizing lotion, essence lotion and the like, and the formula of the raw materials is as follows:

the preparation process of the water-based product of this example is as follows:

(1) adding glycerol and butanediol into a water phase kettle, adding sodium hyaluronate, stirring at 300rpm, adding water and disodium ethylene diamine tetraacetate into the kettle, heating to 85 deg.C, and dissolving to clarify to obtain water phase.

(2) Taking the natural antibacterial agent obtained in the embodiment 1 of the invention, PEG-40 hydrogenated castor oil, 1, 2-octanediol and 1, 2-pentanediol, placing into an oil phase kettle, uniformly stirring, and heating to 85 ℃ to obtain an oil phase.

(3) Adding the oil phase (2) into the water phase (1), rapidly stirring (at 300rpm) for 5min, and cooling to 45 ℃.

(4) Adding collagen and glucose into the mixture obtained in the step (3), stirring at the speed of 300rpm, and cooling to room temperature.

Comparative example 3 (use of the antibacterial agent prepared in comparative example 1 of the present invention as a preservative in a cream-type product)

The cream product of the comparative example can be used as moisturizing cream, repairing cream and the like, and the formula of the raw materials is as follows:

the preparation process of the cream product of this comparative example is as follows:

(4) The natural antibacterial agent obtained in comparative example 1, collagen, glucose and 1, 2-octanediol were added to (3), and the mixture was stirred at 500rpm and cooled to room temperature.

Comparative example 4 (use of the antibacterial agent prepared in comparative example 2 of the present invention as a preservative in a cream-type product)

(4) The natural antibacterial agent obtained in comparative example 2, collagen, glucose and 1, 2-octanediol were added to (3), and the mixture was stirred at 500rpm and cooled to room temperature.

COMPARATIVE EXAMPLE 5 (Water-based products conventional in the art)

The water product of the comparative example can be used as skin lotion, moisturizing lotion, essence lotion and the like, and the formula of the raw materials is as follows:

the preparation process of the water-based product of this comparative example is as follows:

(4) Adding the tricaprin, collagen, glucose and 1, 2-octanediol into the mixture (3), stirring at 500rpm, and cooling to room temperature.

Effect example 2 (Corrosion prevention challenge test of examples 5 to 7 and comparative examples 3 to 5)

(1) Mode and number of preservative challenge inoculations:

mixed bacteria are adopted for inoculation. Inoculating Staphylococcus aureus, Escherichia coli and Pseudomonas aeruginosa into the sample to be tested until the inoculation quantity is 10⁵～10⁶CFU/g or 10⁵～10⁶CFU/ml; the inoculation amount of Candida albicans and Aspergillus niger was 10⁴CFU/g or 10⁴CFU/ml, three replicates each, were blanked with saline.

(2) Corrosion protection challenge separation assay

The inoculated sample is separated and detected at a specific time: 0 hour, 7 days, 14 days, 21 days and 28 days. After sampling, 100. mu.L of the suspension was diluted with physiological saline and plated on TSB and SDB medium plates, and colonies were counted after culturing at 28 ℃. + -. 1 ℃ or 36 ℃. + -. 1 ℃, as required for the experiment.

(3) Criteria (according to the society for cosmetics and fragrance perfumes (CTFA) standard)

The requirement was that on day 7, bacteria were reduced by 99.9%, aspergillus niger and candida albicans by 90% each, and continued to decline over 28 days, passing the preservative challenge test.

The preservative effect was considered excellent if the average of the number of microorganisms in any one of the three replicates inoculated with the mixed strain fell to below 100CFU/g (CFU/ml) at day 7 and all fell to 0CFU/g (CFU/ml) or < 10CFU/g (CFU/ml) within 28 days.

TABLE 30 hours preservative challenge results

Table 448 hours preservation challenge results

TABLE 57 day preservative challenge results

Table 614 day preservative challenge results

TABLE 721 day preservative challenge results

Table 828 days preservative challenge results

TABLE 9 summary of preservative challenge results

Effect example 3 (test for human skin Patch in examples 5 to 7 and comparative example 4)

(1) The spot test method comprises the following steps: selecting a qualified spot tester, applying about 0.020 g-0.025 g of a test object into the spot tester by a closed spot test method, applying an external special adhesive tape to the curved side of the forearm of a subject, removing the test object after 24 hours, observing skin reaction 30min after the spot tester of the test object is removed, and recording the result according to the skin reaction grouping standard in the cosmetic hygiene Specification (2007 edition).

(2) And (4) evaluating the results: the results are reported in Table 10 (grouping criteria for adverse skin reactions), and are shown in Table 11.

TABLE 10 grouping criteria for adverse skin reactions

TABLE 11 test results of the sample of the example on the patch on human skin

And (4) conclusion: the results of the human skin patch test of each group show that: in 3 of the examples, 30 subjects showed all negative reactions on the skin, while in comparative example 4, 7 subjects showed suspicious reactions and only faint erythema (score scale: grade 0). According to the judgment standard of the skin patch test of human body in the cosmetic hygiene code (2007 edition), 3 groups of examples have no adverse skin reaction on human body, while the comparative example 4 is suspicious reaction, which shows the significance brought by adding hydrolyzed ginsenoside into the raw materials of the invention, namely the safety of the preservative can be greatly improved.

Claims

1. The preparation method of the plant-derived broad-spectrum antibacterial agent is characterized in that raw materials of the plant-derived broad-spectrum antibacterial agent comprise a material A, water, lipase and a pH regulator, wherein the material A comprises the following components in parts by mass of 100: 0.1-5 parts of litsea cubeba oil, 0.1-5 parts of patchouli oil, 0.1-5 parts of citronella oil, 0.1-5 parts of hydrolyzed ginsenoside and 80-99.6 parts of caprylic capric triglyceride; the preparation method comprises the following steps:

2. The preparation method of claim 1, wherein the material A comprises the following components in parts by mass of 100 parts: 0.2-1 part of litsea cubeba oil, 0.2-1 part of patchouli oil, 0.2-1 part of citronella oil, 0.2-1 part of hydrolyzed ginsenoside and 96-99.2 parts of caprylic capric triglyceride.

3. The preparation method according to claim 1, wherein the water is deionized water, and the amount of the deionized water is 1/10 to 1/5 of the amount of the material A; the pH regulator is one or more of sodium hydroxide, sodium carbonate, sodium bicarbonate, hydrochloric acid and citric acid.

4. The method according to claim 1, wherein in the step (2), the amount of the lipase is an amount of the lipase required for adjusting the enzymatic activity of the system after the water and the lipase are mixed to 10 to 500 u/g; the enzymolysis time is more than 1 hour; the stirring speed is 100-300 rpm.

5. The method according to claim 4, wherein in the step (2), the lipase is used in an amount required for adjusting the enzyme activity to 50 to 100 u/g; the enzymolysis time is 1-4 hours; the stirring rate was 200 rpm.

6. The preparation method according to claim 1, wherein in the step (3), the frequency of the ultrasound is 25000 to 35000Hz, and the time of the ultrasound is 2 to 10 minutes; the oil-water separation operation is standing; in the step (4), the aging temperature is-6 ℃ to-2 ℃, and the aging time is 8-14 hours.

7. The method according to claim 6, wherein in the step (3), the frequency of the ultrasound is 30000 Hz; the standing time is 0.5-4 h; in the step (4), the aging temperature is-4 ℃, and the aging time is 12 hours.

8. A broad-spectrum plant-derived antibacterial agent prepared by the preparation method according to any one of claims 1 to 7.

9. Use of the plant-derived broad-spectrum antibacterial agent of claim 8 as a preservative in cosmetics for daily use and/or pharmaceuticals for external use on skin.