CN109966223B - Natural plant-derived preservative and preparation method and application thereof - Google Patents

Abstract

The invention belongs to the field of natural preservatives, and discloses a natural plant-derived preservative as well as a preparation method and application thereof. The natural plant source preservative comprises the following components in percentage by mass: 10-25% of stemona sessilifolia extract, 10-25% of eleuthium japonicum extract, 10-25% of litsea cubeba extract, 10-25% of murraya jasminorage extract and 30-60% of solvent. The natural plant-derived antiseptic is prepared by supercritical CO extraction from 4 kinds of plants including folium Et cacumen Murrayae, radix Stemonae, herba Alii Fistulosi of Iridaceae, and fructus Litseae₂The plant composite extract obtained by extraction has a synergistic effect, widens the antibacterial spectrum of a single plant, has a wider antibacterial spectrum, and can simultaneously and effectively inhibit microorganisms such as escherichia coli, pseudomonas aeruginosa, staphylococcus aureus, bacillus subtilis, candida albicans, aspergillus niger and the like, so that the plant composite extract can be applied to the field of cosmetics.

Description

Natural plant-derived preservative and preparation method and application thereof

Technical Field

The invention belongs to the field of natural preservatives, and particularly relates to a natural plant-derived preservative as well as a preparation method and application thereof.

Background

The appeal of contemporary consumers to cosmetics is "safe, effective, mild". In order to meet the demand, a large amount of natural and organic nutrients and moisture are required to be added into the cosmetics, and the high-nutrient additives create more convenient conditions for the propagation of microorganisms, and can cause the cosmetics to be rotten and deteriorated. Therefore, it is necessary to add a certain amount of preservative to cosmetics to inhibit the metabolism of bacteria, molds and yeasts and to influence their growth and reproduction.

With the advent of the consumer appeal of "natural, organic, preservative free", traditional chemically synthesized preservatives that have certain toxicities due to structure and properties have been gradually replaced by safer preservatives. For example, the imidazolidinyl urea series, the hydantoin derivatives dmdmdmh (formaldehyde donor), isothiazolinone, parabens (paraben), bronopol, quaternary ammonium salt-15, kaempon-CG, triclosan and the like, have good antiseptic effect and low cost, but have large skin irritation, and some of them are even carcinogenic and allergenic, and thus have gradually become replaced by new natural preservatives in cosmetics and personal care products. Most of the reported natural preservatives are derived from natural organisms, including animal resources, plant resources, microorganisms and the like. For example, ZL 201010564574.3 discloses a natural cosmetic antibacterial ingredient containing carboxymethyl chitosan quaternary ammonium salt as a main ingredient; ZL200810138204.6 discloses a natural preservative of recombinant crocodile antibacterial peptide, ZL 201410315332.9, ZL 201410314113.9 and ZL 201410314114.3 respectively disclose a series of natural preservatives of Chinese alligator antibacterial peptide Alligatorin; CN201810153934.7 discloses a microbial preservative of streptomyces albus fermentation; another large class of preservatives is derived from the compounding of natural plant antibacterial components, such as compound plant extracts related to patents CN 201810634473.5, CN201810634645.9, CN201410850206.3, CN201810634435.X, CN201711442019.1 and the like. Because different kinds of plant extracts have respective antibacterial spectrum and application range, and a single component cannot achieve the comprehensive anticorrosion effect, the synergistic effect of multiple natural antibacterial components is often needed to achieve the broad-spectrum anticorrosion capability.

At present, natural preservatives really applied to cosmetics are not many, and the main reason is that the preservative capability is limited, or the bacteriostatic effect on a single strain is obvious, but the broad-spectrum bacteriostatic effect cannot be achieved. Therefore, how to screen proper plant antiseptic functional components and how to match different antibacterial components to achieve synergistic antibacterial effect by adopting any extraction process is the key to develop a novel natural antiseptic.

Disclosure of Invention

In order to overcome the problem of great skin irritation caused by the chemically synthesized preservative in the prior art, the invention aims to provide a natural plant-derived preservative.

The invention also aims to provide a preparation method of the natural plant-derived preservative.

The invention further aims to provide application of the natural plant-derived preservative in cosmetics.

The purpose of the invention is realized by the following scheme:

a natural plant source preservative is composed of the following components in percentage by mass:

preferably, the natural plant-derived preservative comprises the following components in percentage by mass:

more preferably, the natural plant-derived preservative comprises the following components in percentage by mass:

the solvent is polyhydric alcohol, preferably at least one of 1, 3-propylene glycol, 1, 2-propylene glycol, butanediol, glycerol and dipropylene glycol; and the polyols can be obtained by biological extraction or fermentation.

The radix Stemonae extract, herba Alii Fistulosi extract of Iridaceae, fructus Litseae extract, and folium Et cacumen Murrayae extract are processed by supercritical CO extraction₂Is prepared by an extraction technology;

the stemona sessilifolia extract is prepared by the following method: drying the stemona sessilifolia leaves by using a blast drier at 40-60 ℃, crushing, sieving with a 40-80 mesh sieve to obtain stemona sessilifolia leaf powder, and adding supercritical CO into the powder₂Extracting kettle of extractor for CO₂Performing supercritical extraction to obtain green oily radix Stemonae extract, and refrigerating at 4 deg.C for use.

The extract of the red onion of the Iridaceae family is prepared by the following method: drying eleutherine onion in Iridaceae at 40-60 deg.C by air-blast dryer, pulverizing, sieving with 40-80 mesh sieve to obtain eleutherine onion powder, and adding supercritical CO into the powder₂Extracting kettle of extractor for CO₂Performing supercritical extraction to obtain red brown oily extract of red onion of Iridaceae, and refrigerating at 4 deg.C.

The litsea cubeba extract is prepared by the following method: drying litsea cubeba fruits at 40-60 ℃ by using a blast drier, crushing, sieving with a 40-80 mesh sieve to obtain litsea cubeba fruit powder, and adding supercritical CO into the powder₂Extracting kettle of extractor for CO₂Performing supercritical extraction to obtain brown oily fructus Litseae extract, and refrigerating at 4 deg.C.

The murraya jasminorage extract is prepared by the following steps: drying the Murraya paniculata flowers at 40-60 ℃ by using a blast dryer, crushing, sieving with a 40-80 mesh sieve to obtain Murraya paniculata flower powder, and adding supercritical CO into the powder₂Extracting kettle of extractor for CO₂Performing supercritical extraction to obtain brown yellow oily folium Et cacumen Murrayae extract, and refrigerating at 4 deg.C.

The supercritical CO₂The extraction conditions were: with CO₂Is supercritical fluid, the extraction pressure is 20-50 MPa, the extraction temperature is 20-60 ℃, and CO is added₂The flow rate is 10-200 g/min, the extraction time is 1-5 h, and the oily extract in the separation kettle is collected.

Preferably, the supercritical CO is₂The radix Stemonae extract is extracted under the conditions of extraction pressure of 25MPa, extraction temperature of 52 deg.C, and CO₂The flow rate was 100g/min and the extraction time was 1.0 h.

Preferably, the supercritical CO is₂The conditions for extracting the red onion extract comprise extraction pressure of 35MPa, extraction temperature of 35 deg.C, and CO₂The flow rate was 60g/min and the extraction time was 2.0 h.

Preferably, the supercritical CO is₂The conditions for extracting the litsea cubeba extract comprise that the extraction pressure is 25MPa, the extraction temperature is 50 ℃, and CO is added₂The flow rate was 120g/min and the extraction time was 1.0 h.

Preferably, the supercritical CO is₂The extraction conditions of the Murraya koenigii extract comprise extraction pressure of 31MPa, extraction temperature of 25 deg.C, and CO₂The flow rate was 20g/min and the extraction time was 1.5 h.

The preparation method of the natural plant source preservative comprises the following steps:

mixing radix asparagi officinalis extract, herba Alii Fistulosi extract of Iridaceae, fructus Litseae extract, and folium Et cacumen Murrayae extract according to formula to obtain composite plant extract, dissolving the composite plant extract in solvent, stirring, and membrane filtering to obtain the natural plant-derived antiseptic.

In the preparation method, the stirring is performed to fully and uniformly mix the raw materials, and the conventional stirring speed in the field can be realized, so that the stirring speed and the stirring time are not required to be limited. Preferably, the stirring speed is 500-1500 r/min, and the stirring time is 20-60 min.

The membrane in the membrane filtration is a microfiltration membrane with the aperture of 0.8-1.5 mu m.

The natural plant-derived antiseptic is prepared by supercritical CO extraction from 4 kinds of plants including folium Et cacumen Murrayae, radix Stemonae, herba Alii Fistulosi of Iridaceae, and fructus Litseae₂The plant composite extract obtained by extraction has a synergistic effect, widens the antibacterial spectrum of a single plant, has a wider antibacterial spectrum, and can simultaneously and effectively inhibit microorganisms such as escherichia coli, pseudomonas aeruginosa, staphylococcus aureus, bacillus subtilis, candida albicans, aspergillus niger and the like, so that the plant composite extract can be applied to the field of cosmetics.

Compared with the prior art, the invention has the following advantages and beneficial effects:

(1) the natural preservative prepared by the invention provides a preservative compounded by four plant extracts, has a good bacteriostatic effect and a wide antibacterial spectrum, and can effectively inhibit microorganisms such as escherichia coli, pseudomonas aeruginosa, staphylococcus aureus, bacillus subtilis, candida albicans and the like.

(2) The invention adopts supercritical fluid extraction technology to extract bacteriostatic active ingredients from stemona sessilifolia, litsea cubeba, murraya paniculata and eleutherine, and the obtained extract has strong bacteriostatic and antiseptic capabilities.

(3) The natural preservative does not contain conventional chemical synthetic substances, and has the characteristics of low skin irritation, safety and mildness.

Detailed Description

The present invention will be described in further detail with reference to examples, but the embodiments of the present invention are not limited thereto.

The reagents used in the examples are commercially available without specific reference.

Examples

(1) Preparation of radix stemonae extract

Drying the cleaned sessile stemona leaves at 60 ℃ by using a blast drier, crushing by using a mechanical crusher, and then sieving by using a 60-mesh sieve to obtain sessile stemona leaf powder. Accurately weighing 10.0kg radix Stemonae powder, and adding supercritical CO₂Extracting kettle of extractor for CO₂And (4) performing supercritical extraction. Supercritical CO₂Extracting radix Stemonae with CO at extraction temperature of 52 deg.C under extraction pressure of 25MPa to obtain antibacterial component₂The flow rate was 100g/min, the extraction time was 1.0h, 632g of green oil-like extract in the separation vessel was collected, and the extraction yield was 6.32%. The sample was placed in a 4 ℃ freezer for use.

(2) Preparation method of extract of Allium fistulosum Bunge of Iridaceae

Drying the cleaned red onion at 50 ℃ by using a blast drier, crushing by using a mechanical crusher, and sieving by using a 50-mesh sieve to obtain red onion powder. Accurately weighing 10.0kg of scallion powder, and adding supercritical CO₂Extracting kettle of extractor for CO₂And (4) performing supercritical extraction. Supercritical CO₂Extracting antibacterial components from herba Alii Fistulosi by 35MPa at 35 deg.C with CO₂The flow rate is 60g/min, the extraction time is 2.0h, 786g of the reddish brown oily extract in the separation kettle is collected, and the extraction rate is 7.86%. The sample was placed in a 4 ℃ freezer for use.

(3) Preparation of litsea cubeba extract

Drying the cleaned litsea cubeba fruits at 55 ℃ by using a forced air dryer, crushing by using a mechanical crusher, and sieving by using a 40-mesh sieve to obtain litsea cubeba powder. Accurately weighing 10.0kg of Litsea cubeba powder, and adding supercritical CO₂Extracting kettle of extractor for CO₂And (4) performing supercritical extraction. Supercritical CO₂Method for extracting litsea cubeba to inhibit bacteriaThe extraction process comprises the steps of extracting at the temperature of 50 ℃ under the pressure of 25MPa and CO₂The flow rate was 120g/min, the extraction time was 1.0h, 3130g of the yellowish brown oily extract from the separation vessel was collected, and the extraction rate was 31.30%. The sample was placed in a 4 ℃ freezer for use.

(4) Preparation of Murraya koenigii extract

Drying the cleaned Murraya paniculata flowers at 55 ℃ by using a blast drier, crushing by using a mechanical crusher, and sieving by using a 50-mesh sieve to obtain Murraya paniculata powder. Accurately weighing 10.0kg of folium Et cacumen Murrayae powder, and adding supercritical CO₂Extracting kettle of extractor for CO₂And (4) performing supercritical extraction. Supercritical CO₂Extracting folium Et cacumen Murrayae with CO at 25 deg.C and 31MPa to obtain antibacterial component₂The flow rate is 20g/min, the extraction time is 1.5h, 510g of brown yellow oily extract in the separation kettle is collected, and the extraction rate is 5.10%. The sample was placed in a 4 ℃ freezer for use.

The results of qualitative analysis of the chemical components contained in the four plant extracts are shown in table 1:

TABLE 1 qualitative analysis of chemical components of four plant extracts

Note: "+" is a positive reaction; "-" is a negative reaction; s is an extract of radix stemonae sessilifoliae; e is extract of Allium fistulosum of Iridaceae; l is fructus Litseae extract; m is folium Et cacumen Murrayae extract.

The results of qualitative analysis of chemical components of the 4 plant extracts are shown in table 1, and therefore, the 4 plant extracts contain flavonoids, phenols, tannins and organic acids. In addition, S (radix Stemonae extract) also contains alkaloid and saponin; e (extract of herba Alii Fistulosi of Iridaceae) also contains anthraquinones and steroids; l (Litsea cubeba extract) also contains saponins; m (Murraya koenigii extract) also contains alkaloids.

(5) Preparation of natural plant source preservative

Mixing the obtained radix Stemonae extract, herba Alii Fistulosi extract of Iridaceae, fructus Litseae extract, and folium Et cacumen Murrayae extract at a certain ratio, dissolving the obtained compound plant extract in a certain amount of butanediol solvent, stirring at 800r/min for 30min, and filtering with 0.8 μm microfiltration membrane to obtain natural antiseptic. The compounding ratios of the respective components in examples 1 to 4 and comparative examples 1 to 5 are shown in Table 2.

TABLE 2 composition ratio of different natural plant-derived preservatives

The bacteriostatic activity of the botanical preservatives of examples 1-4 and comparative examples 1-5 was evaluated by measuring the zone of inhibition by agar plate punching. Escherichia coli (Escherichia coli) ATCC8739, Pseudomonas aeruginosa (Pseudomonas aeruginosa) ATCC9027, Staphylococcus aureus (Staphylococcus aureus) ATCC 6538; the yeasts Candida albicans (Candida albicans) ATCC10231 and the mould Aspergillus niger (Aspergillus niger) ATCC16404 were the test strains, and the samples were tested for bacteriostatic activity. Diluting the above strains to a concentration of about 10 with sterile physiological saline⁶CFU/ml bacterial suspension. Preparing a bacterial culture medium: lecithin tween 80-nutrient agar medium, mycoderm medium: potato dextrose agar medium, which are all media conventionally used in the art, but the present invention is not limited to such media. Sterilizing the culture medium at 120 deg.C for 15min, cooling to 50 deg.C, pouring into a plate with diameter of 90mm, shaking, and condensing; respectively adding 100 μ l of the above bacterial suspension into corresponding bacteria culture medium or mold culture medium (such as bacterial suspension is cultured on bacteria culture medium), and uniformly coating with a coating rod for use. And (3) punching the prepared bacterium-carrying flat plate by using a sterile puncher with the diameter of 6mm under the aseptic condition, picking out the agar block by using sterilized tweezers, adding 50 mu l of the products of the examples 1-4 and the comparative examples 1-5 of the invention into the hole, culturing in a constant-temperature incubator, observing the bacteriostatic effect after the culture is finished, and measuring the diameter of a bacteriostatic ring (the diameter unit of the bacteriostatic ring is mm). Three replicates were performed and averaged, with the results shown in table 3 below:

TABLE 3 results of zone of inhibition experiments for examples 1 to 4 and comparative examples 1 to 5

As can be seen from Table 3, the compounded plant-derived preservative has broad antibacterial spectrum under the same concentration, and the antibacterial effect of the examples 1-3 is obviously better than that of the comparative examples 1-4 which are single plant extracts of radix stemonae sessilifoliae extract, iridaceae red onion extract, litsea cubeba extract and murraya jasminorage extract, and the comparative example 5 which is butanediol solvent. In addition, the bacteriostatic effect of the embodiment 1 is better than that of the embodiments 2 and 3, which shows that the preservative effect of the four plant extracts with uniform content is better than that of the preservative with more content of one or more components and non-uniform components; the bacteriostatic effect of the examples 1-3 is obviously better than that of the example 4, which shows that the less the content of the solvent, the more the effective components of the plant extract are, and the better the bacteriostatic effect of the compound plant-derived preservative is.

Application examples

Application in cosmetics, cosmetics A-E shown in Table 4 were prepared.

TABLE 4 cream formulations of different preservatives

The preparation process of the cosmetics A-E comprises the following steps:

(1) mixing the components of the phase A, heating to 75-85 ℃, and uniformly stirring for later use;

(2) mixing the phase B components, heating to 75-85 ℃ until the components are melted, and uniformly stirring for later use;

(3) adding the phase B into the phase A, and homogenizing at 10000-15000 rpm for 3-5 min;

(4) cooling to 50-60 ℃, adding the component C, and uniformly stirring for later use;

(5) cooling to 40-45 ℃, adding the butyl component, and uniformly stirring;

(6) the pH (10% aqueous solution) of the formulations A to E is controlled to 5.5 to 7.0.

The 10% aqueous solution in the step (6) means that the formula products A-E are dissolved in the aqueous solution, and the mass fraction of the formula products A-E is 10%.

And (3) testing the efficacy: according to the united states pharmacopeia (21 st edition) microbial preservative efficacy test, microbial species: the bacteria are mixed strains of Escherichia coli (ATCC8739), Pseudomonas aeruginosa (ATCC9027) and Staphylococcus aureus (ATCC 6538); the yeast and mold are mixed strains of Candida albicans (ATCC10231) and Aspergillus niger (ATCC 16404). The detection method comprises weighing 30g of each test sample, adding mixed bacteria or mixed mold suspension to obtain final bacteria content of 5.0 × 10/g⁶3.0X 10 of a bacterium⁵And (3) mold. Mixing, and culturing at 28 deg.C. Samples were taken for analysis on days 0, 7, 14, 21 and 28 of inoculation: accurately weighing 3g of sample in a sterilized conical flask, adding 27ml of sterilized normal saline, and fully and uniformly oscillating, wherein the suspension is a diluent of 1: 10; then sequentially diluting the mixture by 10 times with sterilized normal saline. The bacteria content of the test sample was counted by plate pour method, the bacteria culture was carried out at 37 ℃ for 3 days, and the mold culture was carried out at 28 ℃ for 5 days. The cosmetics A-E are subjected to a microbial corrosion prevention challenge experiment, and the judgment standard is as follows: after the sample is inoculated with a certain amount of mixed strains, the amount of viable bacteria is reduced to not more than 0.1% of the initial concentration (less than or equal to 0.1% of the initial concentration) on the 14 th day, and the amount of viable bacteria is continuously reduced from 14 days to 28 days. Compliance with the standard is preservative efficacy and non-compliance is preservative inefficiency. The test results are shown in table 5 below.

TABLE 5 cosmetic 28-day preservative challenge test results for different formulations

As can be seen from table 5, the natural preservative prepared in example 1 has good performance, and the preservative effect of the natural plant-derived preservative with 1.2% of the addition amount is equivalent to that of 1% of euxyl PE 9010 (0.9% phenoxyethanol and 0.1% ethylhexyl glycerin), and is obviously better than that of the preservative containing "no addition" of polyols. Therefore, the natural preservative prepared in example 1 can replace part of synthetic preservatives, so that the formula has milder and safer effects.

The influence of different pH values on the preservative ability was investigated: using cosmetic E as a sample, cosmetic E adjusted to pH shown in table 6 using sodium hydroxide, lactic acid, or citric acid was named cosmetic F when pH was 5.0, cosmetic E when pH was 5.5 was named cosmetic G, cosmetic E when pH was 6.0 was named cosmetic H, cosmetic E when pH was 7.0 was named cosmetic I, and cosmetic E when pH was 8.0 was named cosmetic J.

TABLE 6 pH of different cosmetics

	Cosmetic F	Cosmetic G	Cosmetic H	Cosmetic I	Cosmetic J
						pH	5.0	5.5	6.0	7.0	8.0

The preservative challenge results for cosmetics F-J are shown in table 7:

table 7 results of 28 day preservative challenge experiments for different pH cosmetics

As can be seen from Table 7, when the natural plant-derived preservative prepared in example 1 of the present invention is used, the preservative effect can be maintained well in the range of pH 5.0-8.0, and the cosmetic formulations F-J can pass the preservative challenge experiment.

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims (4)

1. A natural plant source preservative is characterized by comprising the following components in percentage by mass:

the stemona sessilifolia extract is prepared by the following method: drying the stemona sessilifolia leaves by using a blast drier at 40-60 ℃, crushing, sieving with a 40-80 mesh sieve to obtain stemona sessilifolia leaf powder, and adding supercritical CO into the powder₂Extracting kettle of extractor for CO₂Performing supercritical extraction to obtain green oily radix Stemonae extract, and refrigerating at 4 deg.C;

the extract of the eleutherine onion is prepared by the following steps: drying Allium fistulosum Bunge of Iridaceae at 40-60 deg.C by using air-blast dryer, pulverizing, sieving with 40-80 mesh sieve to obtain Allium fistulosum Bunge powderThen adding the powder into supercritical CO₂Extracting kettle of extractor for CO₂Performing supercritical extraction to obtain red brown oily extract of red onion of Iridaceae, and refrigerating at 4 deg.C;

the litsea cubeba extract is prepared by the following method: drying litsea cubeba fruits at 40-60 ℃ by using a blast drier, crushing, sieving with a 40-80 mesh sieve to obtain litsea cubeba fruit powder, and adding supercritical CO into the powder₂Extracting kettle of extractor for CO₂Performing supercritical extraction to obtain brown oily fructus Litseae extract, and refrigerating at 4 deg.C;

the murraya jasminorage extract is prepared by the following steps: drying the Murraya paniculata flowers at 40-60 ℃ by using a blast dryer, crushing, sieving with a 40-80 mesh sieve to obtain Murraya paniculata flower powder, and adding supercritical CO into the powder₂Extracting kettle of extractor for CO₂Performing supercritical extraction to obtain brown yellow oily folium Et cacumen Murrayae extract, and refrigerating at 4 deg.C;

the supercritical CO₂The radix Stemonae extract is extracted under the conditions of extraction pressure of 25MPa, extraction temperature of 52 deg.C, and CO₂The flow rate is 100g/min, and the extraction time is 1.0 h;

the supercritical CO₂The conditions for extracting the red onion extract comprise extraction pressure of 35MPa, extraction temperature of 35 deg.C, and CO₂The flow rate is 60g/min, and the extraction time is 2.0 h;

the supercritical CO₂The conditions for extracting the litsea cubeba extract comprise that the extraction pressure is 25MPa, the extraction temperature is 50 ℃, and CO is added₂The flow rate is 120g/min, and the extraction time is 1.0 h;

the supercritical CO₂The extraction conditions of the Murraya koenigii extract comprise extraction pressure of 31MPa, extraction temperature of 25 deg.C, and CO₂The flow rate is 20g/min, and the extraction time is 1.5 h;

the solvent is butanediol.

2. A method for preparing the natural plant-derived preservative according to claim 1, characterized by comprising the steps of:

mixing radix asparagi officinalis extract, herba Alii Fistulosi extract of Iridaceae, fructus Litseae extract, and folium Et cacumen Murrayae extract according to formula to obtain composite plant extract, dissolving the composite plant extract in solvent, stirring, and membrane filtering to obtain the natural plant-derived antiseptic.

3. The method for producing a natural plant-derived preservative according to claim 2, characterized in that:

the membrane in the membrane filtration is a microfiltration membrane with the aperture of 0.8-1.5 mu m.

4. Use of the natural plant-derived preservative according to claim 1 in the cosmetic field.