CN115154404A

CN115154404A - Plant composition with tightening and anti-aging functions and preparation method and application thereof

Info

Publication number: CN115154404A
Application number: CN202211081510.7A
Authority: CN
Inventors: 黄坤洋; 徐健; 曹畅达; 王利芬; 刘新琳; 陶宏兵; 邓小锋; 谢小保
Original assignee: Long Yue Industrial Guangzhou Co ltd; Guangdong Demay New Materials Technology Co ltd
Current assignee: Long Yue Industrial Guangzhou Co ltd; Guangdong Demay New Materials Technology Co ltd
Priority date: 2022-09-06
Filing date: 2022-09-06
Publication date: 2022-10-11
Anticipated expiration: 2042-09-06
Also published as: CN115154404B

Abstract

The invention discloses a plant composition with tightening and anti-aging functions as well as a preparation method and application thereof, wherein the preparation method comprises the following steps: respectively pretreating fresh plants, sieving to obtain plant powder, and measuring microorganism, heavy metal and pesticide residue; respectively adding the qualified plant powder into the extracting solution for pre-extraction, then adding a green eutectic solvent for extraction, and filtering to obtain filtrate and filter residue; carrying out ultrafiltration on the filtrate to obtain a plant extract A; mixing the filter residue with water, adding Bacillus bifidus, fermenting, adding lysozyme, and collecting supernatant to obtain fermentation filtrate B; mixing the plant extract A corresponding to each plant with the fermentation filtrate B, adding antiseptic to obtain plant extract, mixing the plant extracts, and adding solvent and antiseptic to obtain plant composition with high active ingredient content, tightening and antiaging effects, skin moisture content increasing, collagen ColI expression promoting, and free radical scavenging effects.

Description

Plant composition with tightening and anti-aging functions and preparation method and application thereof

Technical Field

The invention relates to the technical field of cosmetics, in particular to a preparation method of a plant composition with tightening and anti-aging functions, the plant composition with tightening and anti-aging functions prepared by the preparation method and application of the plant composition in cosmetics.

Background

The skin is the organ with the largest contact area between the human body and the outside, maintains the balance in the body, and resists the interference of various external factors, such as ultraviolet rays, active oxygen, heavy metals and the like. The influence of these external factors directly leads to irreversible aging of the skin. Aging of the face inevitably becomes painful in many people's mind as people age. The aging degree of facial skin, subcutaneous tissue and muscle is gradually developed, and mainly shows that the papillary layer of dermis is thinned, elastic fibers are broken, collagen fibers are denatured, hyaluronic acid in the interlayer is reduced, the subcutaneous tissue is atrophied, the structure of deep soft tissue is loosened, the skin is dry and loses the water property, age pigment is aged and the like. In order to reduce aging caused by external factors, the product with the effects of tightening and resisting aging can be smeared on the epidermis of a human body, and the skin is effectively protected.

At present, most plant extract for tightening skin is extracted directly by a solvent, although the extraction method is simple to operate, the extraction efficiency is low, the extract contains simple and trace plant active ingredients, and most of the extract is micromolecule and polysaccharide compounds which can be directly dissolved in the solvent. For these reasons, the extract is suitable for cosmetics, and can achieve good effects of tightening skin and resisting aging mostly only with high addition amount, and the high addition amount greatly improves the cost of the cosmetics.

Disclosure of Invention

One of the purposes of the invention is to provide a preparation method of a plant composition with the functions of tightening and resisting aging and the plant composition prepared by the preparation method, and the plant composition which has high active ingredient content, tightening and resisting aging, and has the functions of improving the moisture content of skin, promoting the expression of collagen ColI, promoting the removal of free radicals and improving cell aging can be prepared by the preparation method.

The invention also aims to provide application of the plant composition with the functions of tightening and resisting aging in cosmetics.

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

a preparation method of a plant composition with tightening and anti-aging effects is provided, which comprises the following steps:

s10, respectively pretreating fresh peach trees, rhizobia, arabic trees, buddleja officinalis, water lily and rosemary herbs, sieving to obtain plant powder, and determining microorganisms, heavy metals and pesticide residues in the plant powder;

s20, respectively adding the qualified plant powder into the extracting solution for pre-extraction, then adding a green eutectic solvent for extraction, and filtering to obtain filtrate and filter residue;

s30, carrying out ultrafiltration treatment on the filtrate to obtain a plant extract A; mixing the filter residue with water, adding Bacillus bifidus, fermenting, adding lysozyme, breaking strain, and collecting supernatant to obtain fermentation filtrate B containing strain lysate;

s40, mixing plant extract A corresponding to each plant with fermentation filtrate B, adding a preservative to respectively prepare peach resin extract, rhizobia gum extract, arabic gum extract, pale butterflybush flower extract, water lily extract and rosemary extract, mixing peach resin extract, rhizobia gum extract, arabic gum extract, pale butterflybush flower extract, water lily extract and rosemary extract, and adding a solvent to prepare the plant composition.

According to the invention, the extract of the butterflybush flower and the rosemary herb enters the stratum corneum basal layer to protect the formed cells and reduce the damage of endogenous free radicals; the water lily and rhizobium glue extracting solution constructs a dimensional and stable structure, has stimulation effect on the generation of collagen and elastin which form protein fibers, and is combined with antioxidant active molecules of basal layer buddleja officinalis and rosemary herb extracting solution to enhance the skin activity; finally, a large amount of biological polysaccharide is provided by the peach resin and the Arabic gum extracting solution, the structure contains a large amount of hydroxyl, the biological polysaccharide can be combined with water in a hydrogen bond mode, and meanwhile, the biological polysaccharide has good film forming property, can form a 'water locking film' on the surface of skin to prevent the skin water loss, and achieves the effect of lasting moisture preservation. The invention constructs triple biological membranes, and forms a semi-permeable closed hydrated biological membrane inside and outside the skin so as to make the skin fine and smooth.

Among them, the green eutectic solvent EDSs is a green harmless extraction solvent formed by two or more solid or liquid substances through hydrogen bond interaction, and is named because its melting point is significantly lower than that of each single component. Compared with the traditional solvent, the extraction rate of polar and nonpolar metabolites is higher by adopting the green eutectic solvent EDSs, and the extraction rate of most of main phenolic compounds from the EDSs is between 75% and 97%.

Wherein, the bifidobacterium is used as a physiological beneficial bacterium and has a plurality of important physiological functions of biological barrier, nutrition, anti-tumor, immunity enhancement, gastrointestinal tract function improvement, aging resistance and the like for human health. The plant extract obtained by fermenting the bifidobacterium can effectively improve the utilization rate of plants.

Peach resin extract is a plant extract that is more commonly used for tightening. Under normal conditions, peach trees do not actively secrete peach resin, and a certain amount of peach resin is not secreted until the peach trees protect the peach trees and accelerate wound healing after the surfaces of the barks of the peach trees are cracked by external force. In this respect, it is understood that there will be a wound in the area where the peach resin exists, and the wound will naturally be infected by mold or other pathogenic bacteria, so that the peach resin is also inevitably contaminated with toxins or pathogenic bacteria. Certainly, pollutants such as heavy metals floating in the air can be contaminated on the peach resin, and the heavy metal indexes are not monitored in the existing extraction technology so as to ensure the safety of the peach resin.

The invention samples and inspects the plant powder, ensures the safety of the raw material and ensures that the raw material does not contain pollutants such as heavy metal and the like. In the extraction process, an eutectic solvent EDSs is selected as an extracting agent, so that the extraction efficiency can be effectively improved, meanwhile, filtered filter residues are subjected to bifidobacterium fermentation, the residual active ingredients are recycled, and the obtained fermentation filtrate is mixed with an extracting solution to prepare a plant fermentation extracting solution (peach resin extracting solution, rhizobium gum extracting solution, gum arabic extracting solution, butterflybush flower extracting solution, water lily extracting solution and rosemary herb extracting solution). The invention selects specific plants as raw materials, and combines the traditional extraction method, the deep eutectic solvent extraction, the fermentation extraction and the lysozyme strain crushing to realize the synergistic interaction, thereby obtaining the plant composition with high active ingredient content and excellent tightening and anti-aging effects.

Further, step S10 specifically includes: drying fresh plants (drying by hot air at 30-60 ℃), respectively crushing at-35-0 ℃, and then sieving by a 200-400-mesh sieve to obtain plant powder. As is known, the traditional Chinese medicine generates a large amount of heat in the crushing process to heat the material, so that high-temperature oxidation, deterioration and loss and segregation of effective components of heat-sensitive substances and bioactive substances in the material can occur, and the performance index of the heat-sensitive material is reduced or most of the heat-sensitive material is ineffective. In order to solve the problem of crushing, the invention adopts a JCWF-25C ultra-micro crusher of Jinjunchen mechanical equipment Limited company, utilizes an ultra-low temperature crushing technology, namely, adopts a cooling method while crushing, and configures a refrigerator with the temperature of being as low as-35 ℃, so that the softened or sticky medicine after temperature rise can be easily crushed, and the invention is beneficial to retaining the biological active ingredients and various nutritional ingredients which are not high temperature resistant, thereby improving the efficacy. By adopting an ultra-low temperature crushing technology, the average particle size of the materials is generally below 10um, the uniformity is improved, the color is consistent, and the powder is fine and smooth. By adopting an ultra-low temperature crushing technology, the purpose of ultrafine crushing of materials with high fiber, heat sensitivity, aromaticity, volatility and viscosity can be achieved.

The microorganisms in the plant powder are measured according to the fifth chapter 'microorganism inspection method' of the technical Specification for safety of cosmetics 2015, and the measurement indexes are as follows: the total number of colonies is less than or equal to 500CFU/g, the total number of mould and microzyme is less than or equal to 100CFU/g, and heat-resistant coliform group bacteria, staphylococcus aureus and pseudomonas aeruginosa can not be detected.

According to the general rule of 1 'physicochemical inspection method' of the technical Specification for safety of cosmetics 2015, chapter 4, the heavy metals in the plant powder are measured, and the measurement indexes are as follows: mercury <1 mg/kg, lead <10 mg/kg, arsenic <2 mg/kg, cadmium <5 mg/kg.

The pesticide residue in the plant powder is tested by adopting an LC-MS/MS instrument, the test index is that the pesticide residue cannot be detected, and the specific test method comprises the following steps:

1. conditions of liquid chromatography

Acquity UPLC HSS T3 chromatography column (1.8 μm,2.1 mm × 100 mm); the flow rate is 0.4 mL/min; the sample size was 1. Mu.L (all samples with acetonitrile as solvent were 0.5. Mu.L); the column temperature is 40 ℃; temperature of the sample chamber: 4. DEG C; mobile phase: a is a salt solution (5 mmol/L of amine acetate + 0.05% formic acid solution), and B is methanol; gradient of mobile phase: b10-10% for 0-1 min; for 2-3 min, B25% -25%; 5-6.5 min, B45-60%; 7.5-8.5 min, B60% -75%; for 9-10 min, B80-90%; for 11-12 min, B90-10%; 12-14 min, B10-10%.

2. Conditions of Mass Spectrometry

Electrospray ionization (ESI): SI +/ESI-ionization mode; capillary voltage: 3.5 kV; temperature of drying gas: 450 ℃; flow rate of drying gas: 900 L/h; taper hole air flow rate: 150 L/h; atomization pressure: 7 Bar; collision airflow rate: 0.15 mL/min.

Further, the plant composition comprises the following components in parts by weight:

5-15 parts of peach resin extract;

2-5 parts of rhizobium glue extracting solution;

3-15 parts of Arabic gum extracting solution;

5-15 parts of butterflybush flower extracting solution;

5-18 parts of water lily extracting solution;

5-15 parts of rosemary herb extract;

20-70 parts of butanediol;

0.2-0.4 part of preservative.

10 parts of peach resin extracting solution;

3 parts of rhizobium glue extracting solution;

8 parts of Arabic gum extracting solution;

10 parts of butterflybush flower extracting solution;

15 parts of water lily extracting solution;

8 parts of rosemary herb extract;

45.7 parts of butanediol;

0.3 part of preservative.

Further, step S20 specifically includes: and (2) measuring the weight ratio of the qualified plant powder to the green eutectic solvent, namely soaking the qualified plant powder in purified water according to the weight ratio of 1 (1 to 10), pre-extracting for 1 hour at the temperature of 20 to 45 ℃, rotating at the speed of 50 to 100r/min, adding the green eutectic solvent with the water proportion of 20% to continuously extract, wherein the weight ratio of the plant powder to the green eutectic solvent is 1 (1 to 10), and filtering to obtain filtrate and filter residue after extraction.

Further, the green eutectic solvent consists of HBA and HBD in a molar ratio of 1;

as one scheme of the invention, the HBA is lactic acid, and the HBD is any one of sodium acetate, ammonium acetate, glycine and DL-malic acid.

In another embodiment of the present invention, the HBA is choline chloride, and the HBD is any one of ethylene glycol, glycerol, oxalic acid, urea, D-glucose, sucrose, lactic acid, and acetamide.

The preparation method of the green eutectic solvent comprises the following steps: HBA and HBD are stirred uniformly at 80 ℃ according to the mol ratio, and then 20% of water is added and mixed to prepare the green eutectic solvent.

Further, step S30 specifically includes the following steps:

s30a, carrying out ultrafiltration treatment on the filtrate to obtain a plant extract A;

s30b, mixing the filter residue with water according to the volume ratio of 1 to 10, and uniformly stirring at the speed of 100-400r/min to prepare a mixed solution;

s30c, transferring the mixed solution into a fermentation tank, adding a cultured mixed bacterial solution of bifidobacterium consisting of bifidobacterium longum, bifidobacterium animalis and bifidobacterium globosum, wherein the concentration of the mixed bacterial solution is 2 x 10 ⁷ Fermenting for 7 to 14 hours at the temperature of 20 to 45 ℃ at the speed of 210r/min by using CUF/ml to prepare fermentation liquor;

s30d, adding lysozyme with the mildew activity of 20000U/mg into the fermentation liquor to crush the strains, wherein the using amount of the lysozyme is 0.1 to 0.3 percent of the fermentation liquor;

s30e, centrifuging the fermentation liquor, and taking supernatant to obtain fermentation filtrate B containing the strain lysate.

Further, in step S40, the preservative is selected from one or more of chlorphenesin, methylparaben, phenoxyethanol, carvone, and hexylene glycol; the solvent is one or more selected from butanediol, propylene glycol, and 1, 3-propylene glycol.

Further preferably, the plant composition with tightening and anti-aging is prepared by the following method:

drying fresh peach tree, rhizobium, arabic tree, butterflybush flower, water lily and rosemary herb, then crushing at low temperature by using a JCWF-25C ultrafine crusher at the temperature of-35 ℃, sieving by using a 300-mesh sieve after crushing, and measuring the microbial, heavy metal and pesticide residues of the prepared plant powder. Soaking qualified plant powder in purified water according to a weight ratio of 1:5, pre-extracting at 35 ℃ for 1 hour at a rotation speed of 60r/min, adding EDSs (choline chloride: glycerol =1: 2) with a water proportion of 20%, continuously extracting for 1 hour, and filtering after extraction is finished to obtain a filtrate A and a filter residue B, wherein the addition of the EDSs is in a weight ratio of 1;

filtering the filtrate A with ultrafilter, controlling the cut-off molecular weight of the filtering membrane at 5000D, and ultrafiltering to obtain plant extractive solution A of various plants. The residue B was mixed with water (v: v = 1) and stirred uniformly at 200r/min in a stirring tank, and the mixture was transferred to a fermentation tank and added with a mixed bacterial solution of cultured bifidobacteria (2 × 10 ⁷ CUF/ml), 210r/min, and fermenting at 31 ℃ for 12 hours. After the fermentation is finished, adding lysozyme with the mildew activity of 20000U/mg for strain crushing, wherein the usage amount of the lysozyme is 0.2 percent of the fermentation liquor. And finally, centrifuging the fermentation liquor at a high speed, and taking supernatant to obtain fermentation filtrate B containing the strain lysate. Mixing the plant extract A corresponding to each plant with the fermentation filtrate B, and adding chlorphenesin 0.15 wt% and methylparaben 0.15 wt% to obtain plant extracts of various plants;

10 parts of peach resin extract, 3 parts of rhizobia gum extract, 8 parts of gum arabic extract, 10 parts of butterflybush flower extract, 15 parts of water lily extract, 8 parts of rosemary extract, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben are mixed according to the parts by weight to prepare the plant composition with the functions of tightening and resisting senility.

In another aspect, a plant composition with compact and anti-aging effects is provided, and is prepared by the preparation method.

In another aspect, the application of the plant composition with the functions of tightening and resisting aging prepared by the preparation method in cosmetics is provided. Such as essence cream, essence milk, essence liquid, etc.

The invention has the beneficial effects that: the invention selects specific plants as raw materials, and combines the traditional extraction method, the deep eutectic solvent extraction, the fermentation extraction and the lysozyme strain crushing to realize the synergistic interaction, thereby obtaining the plant composition with high active ingredient content and excellent tightening and anti-aging effects.

Drawings

FIG. 1 is a graphical representation of the total alkaloid yield from a botanical composition of the invention;

FIG. 2 is a schematic representation of the total sugar content of the plant composition according to the invention;

FIG. 3 is a graph showing the total phenolic content of the botanical compositions of the present invention;

FIG. 4 is a graph of the skin tightening effect of the botanical compositions of the present invention;

FIG. 5 is a graph of moisture values over time for a subject's forearm area (1) painted with a corresponding test sample in accordance with the present invention;

FIG. 6 is a graph of moisture values over time for the forearm area (2) of a subject of the invention after application of a corresponding test sample;

FIG. 7 is a graph of moisture values over time for a subject's forearm area (3) painted with a corresponding test sample in accordance with the present invention;

FIG. 8 is a graph of moisture values over time for a subject's forearm area (4) after application of a corresponding test sample;

FIG. 9 is a graph of moisture values over time for the forearm area (5) of a subject of the invention after application of a corresponding test sample.

Detailed Description

The technical solution of the present invention is further described below by way of specific embodiments.

Unless otherwise specified, various starting materials of the present invention are commercially available or prepared according to conventional methods in the art.

Example 1

Step 1, drying fresh plants (peach tree, rhizobium, arabian tree, buddleja officinalis, water lily and rosemary grass), and carrying out low-temperature grinding by using a JCWF-25C ultrafine grinder of Jinjunchen mechanical equipment limited, wherein the grinding temperature is-35 ℃, sieving by using a 300-mesh sieve, and measuring the residues of microorganisms, heavy metals and pesticides in the prepared plant powder. Soaking qualified plant powder in purified water according to a weight ratio of 1;

and 2, filtering the filtrate A by using an ultrafiltration machine, controlling the cut-off molecular weight of a filtering membrane to be 5000D, and performing ultrafiltration to obtain a plant extracting solution A. Mixing the filter residue B with water (v: v =1: 10), stirring and mixing uniformly at 200r/min by using a stirring tank, transferring to a fermentation tank, and adding a mixed bacterial solution (2 x 10) of cultured bifidobacterium longum, bifidobacterium animalis and bifidobacterium globosum into the fermentation tank ⁷ CUF/ml), 210r/min, and fermenting at 31 ℃ for 12 hours. After the fermentation is finished, adding lysozyme with the mildew activity of 20000U/mg for strain crushing, wherein the usage amount is 0.2% of the fermentation liquor. And finally, centrifuging the fermentation liquor at a high speed, and taking supernatant to obtain fermentation filtrate B containing the strain lysate. Mixing the plant extractive solution A and the fermentation filtrate B, adding 0.15 wt% antiseptic (chlorphenesin and methylparaben), and respectively making into peach resin extractive solution, rhizobia gum extractive solution, gum arabic extractive solution, flos Buddlejae extractive solution, flos Nymphaeae extractive solution and herba Rosmarini officinalis extractive solution;

step 3, mixing the following components in parts by weight: 10 parts of peach resin extract, 3 parts of rhizobium glue extract, 8 parts of gum arabic extract, 10 parts of butterflybush flower extract, 15 parts of water lily extract, 8 parts of rosemary herb extract, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben. Stirring and mixing uniformly by a stirring tank for 60r/min to prepare the plant composition with compact and anti-aging effects.

Example 2

In the preparation method of the plant composition with tightening and anti-aging functions, the

steps

1 and 2 are the same as those in the above example 1, except that the step 3 is to mix and mix the following components in parts by weight: 3 parts of peach resin extract, 3 parts of rhizobium glue extract, 3 parts of gum arabic extract, 5 parts of butterflybush flower extract, 5 parts of water lily extract, 5 parts of rosemary herb extract, 75.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben. Stirring and mixing uniformly by a stirring tank for 60r/min to prepare the plant composition with compact and anti-aging effects.

Example 3

steps

1 and 2 are the same as those in the above example 1, except that the step 3 is to mix and mix the following components in parts by weight: 10 parts of peach resin extract, 5 parts of rhizobium gum extract, 15 parts of gum arabic extract, 15 parts of butterflybush flower extract, 15 parts of water lily extract, 15 parts of rosemary extract, 24.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben. Stirring and mixing uniformly by a stirring tank for 60r/min to prepare the plant composition with compact and anti-aging effects.

Example 4

The plant composition with firming and anti-aging of this example is prepared substantially the same as in example 1 above, except that: the EDSs consist of choline chloride and ethylene glycol in a molar ratio of 1.

Example 5

The plant composition with firming and anti-aging of this example is prepared substantially the same as in example 1 above, except that: the EDSs consist of choline chloride and D-glucose in a molar ratio of 1.

Example 6

The plant composition with firming and anti-aging of this example is prepared substantially the same as in example 1 above, except that: the EDSs are prepared by mixing lactic acid and glycine according to a molar ratio of 3: 1.

Example 7

The plant composition having firmness and anti-aging of this example was prepared substantially the same as in example 1 above, except that: EDSs are prepared by mixing lactic acid and sodium acetate according to a molar ratio of 3: 1.

Example 8

The plant composition with firming and anti-aging of this example is prepared substantially the same as in example 1 above, except that: EDSs are prepared from lactic acid and ammonium acetate in a molar ratio of 3: 1.

Comparative example 1

Step 1, drying fresh plants (peach tree, rhizobia, arabian tree, buddleja officinalis, water lily and rosemary grass), then crushing at low temperature by using a JCWF-25C ultrafine crusher of Jinjunchen mechanical equipment limited, wherein the crushing temperature is-35 ℃, sieving by using a 300-mesh sieve, and measuring the residues of microorganisms, heavy metals and pesticides on the prepared plant powder. Soaking the qualified plant powder in purified water according to a weight ratio of 1.

And 2, filtering the filtrate A by using an ultrafiltration machine, controlling the cut-off molecular weight of a filter membrane to be 5000D, performing ultrafiltration to obtain a plant extracting solution, and adding 0.15 weight percent of preservative (chlorphenesin and methylparaben) to respectively prepare a peach resin extracting solution, a rhizobia gum extracting solution, an Arabic gum extracting solution, a pale butterflybush flower extracting solution, a water lily extracting solution and a rosemary herb extracting solution.

Step 3, mixing the following components in parts by weight: 10 parts of peach resin extract, 3 parts of rhizobium glue extract, 8 parts of gum arabic extract, 10 parts of butterflybush flower extract, 15 parts of water lily extract, 8 parts of rosemary herb extract, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben. Stirring uniformly by a stirring tank at 60r/min to obtain the plant composition with compact and anti-aging effects.

Comparative example 2

And 2, filtering the filtrate A by using an ultrafiltration machine, controlling the cut-off molecular weight of a filtering membrane to be 5000D, and performing ultrafiltration to obtain a plant extracting solution A. Filtering the residueB was mixed with water (v: v =1: 10), stirred and mixed uniformly at 200r/min in a stirring tank, transferred to a fermentation tank, and added with a mixed bacterial solution of cultured bifidobacterium longum, bifidobacterium animalis, and bifidobacterium globosum (2 × 10 ⁷ CUF/ml), 210r/min, and fermenting at 31 ℃ for 12 hours. And (3) after the fermentation is finished, finally, centrifuging the fermentation liquor at a high speed, and taking supernatant to obtain fermentation filtrate B. Mixing the plant extractive solution A and the fermentation filtrate B, adding 0.15 wt% antiseptic (chlorphenesin and methyl hydroxybenzoate) to obtain peach resin extractive solution, rhizobium gum extractive solution, gum arabic extractive solution, flos Buddlejae extractive solution, flos Nymphaeae extractive solution and herba Rosmarini officinalis extractive solution.

Comparative example 3

The plant composition having firmness and anti-aging of this example was prepared substantially the same as in example 1 above, except that: the EDSs were replaced with 70% ethanol solvent.

Comparative example 4

The plant composition with firming and anti-aging of this example is prepared substantially the same as in example 1 above, except that: the EDSs were replaced with propylene glycol solvent.

Comparative example 5

The plant composition with firming and anti-aging of this example was prepared substantially the same as in example 1 above, except that the EDSs were prepared by the following method: the mol ratio of choline chloride: glycerol =1, magnetically stirred in an oil bath pan at 80 ℃ until a homogeneous liquid is formed. Then adding water with the proportion of 30 percent, and mixing to prepare the green eutectic solvent EDSs.

Comparative example 6

The plant composition with firming and anti-aging of this example was prepared substantially the same as in example 1 above, except that the EDSs were prepared by the following method: the mol ratio of choline chloride: glycerol =1, magnetically stirred in an 80 ℃ oil bath until a homogeneous liquid is formed. Then adding water with the proportion of 50 percent, and mixing to prepare the green eutectic solvent EDSs.

Comparative example 7

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the preparation method of the EDSs comprises the following steps: the mol ratio of choline chloride: glycerol =1, magnetically stirred in an 80 ℃ oil bath until a homogeneous liquid is formed. Then adding water with the proportion of 20 percent, and mixing to prepare the eutectic solvent EDSs.

Comparative example 8

The plant composition with firming and anti-aging of this example was prepared substantially the same as in example 1 above, except that the EDSs were prepared by the following method: the molar ratio of choline chloride: glycerol =1, magnetically stirred in an oil bath pan at 80 ℃ until a homogeneous liquid is formed. Then adding water with the proportion of 20 percent, and mixing to prepare the eutectic solvent EDSs.

Comparative example 9

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the preparation method of the EDSs comprises the following steps: the mol ratio of choline chloride: glycerol =2, magnetically stirred in an oil bath pan at 80 ℃ until a homogeneous liquid is formed. Then adding water with the proportion of 20 percent, and mixing to prepare the eutectic extraction solvent EDSs.

Comparative example 10

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the preparation method of the EDSs comprises the following steps: the molar ratio of choline chloride: glycerol =3, magnetically stirred in an oil bath pan at 80 ℃ until a homogeneous liquid is formed. Then adding water with the proportion of 20 percent, and mixing to prepare the eutectic solvent EDSs.

Comparative example 11

The plant composition with firming and anti-aging of this example was prepared substantially the same as in example 1 above, except that the EDSs were prepared by the following method: the molar ratio of choline chloride: glycerol =4, magnetically stirred in an 80 ℃ oil bath until a homogeneous liquid is formed. Then adding water with the proportion of 20 percent, and mixing to prepare the eutectic extraction solvent EDSs.

Comparative example 12 (without peach resin extract, otherwise unchanged)

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 3 parts of rhizobium glue extracting solution, 8 parts of Arabic gum extracting solution, 10 parts of butterflybush flower extracting solution, 15 parts of water lily extracting solution, 8 parts of rosemary herb extracting solution, 55.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 13

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 10 parts of peach resin extract, 8 parts of gum arabic extract, 10 parts of butterflybush flower extract, 15 parts of water lily extract, 8 parts of rosemary herb extract, 48.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 14

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 10 parts of peach resin extract, 3 parts of rhizobium glue extract, 10 parts of butterflybush flower extract, 15 parts of water lily extract, 8 parts of rosemary herb extract, 53.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 15

The preparation method of the plant composition with tightening and anti-aging effects of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 10 parts of peach resin extract, 3 parts of rhizobium glue extract, 8 parts of gum arabic extract, 15 parts of water lily extract, 8 parts of rosemary extract, 55.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

COMPARATIVE EXAMPLE 16 (not containing Nymphaea tetragona extract, otherwise unchanged)

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 10 parts of peach resin extract, 3 parts of rhizobium glue extract, 8 parts of gum arabic extract, 10 parts of butterflybush flower extract, 8 parts of rosemary extract, 60.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 17

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 10 parts of peach resin extracting solution, 3 parts of rhizobia gel extracting solution, 8 parts of gum arabic extracting solution, 10 parts of butterflybush flower extracting solution, 15 parts of water lily extracting solution, 53.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 18

The preparation method of the plant composition with tightening and anti-aging effects of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 5 parts of rhizobium gum extracting solution, 10 parts of Arabic gum extracting solution, 12 parts of butterflybush flower extracting solution, 17 parts of water lily extracting solution, 10 parts of rosemary herb extracting solution, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 19

The preparation method of the plant composition with tightening and anti-aging effects of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 10.6 parts of peach resin extract, 8.6 parts of Arabic gum extract, 10.6 parts of buddleja officinalis extract, 15.6 parts of water lily extract, 8.6 parts of rosemary herb extract, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 20

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 11.6 parts of peach resin extract, 4.6 parts of rhizobium glue extract, 11.6 parts of buddleja officinalis extract, 16.6 parts of water lily extract, 9.6 parts of rosemary herb extract, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 21

The preparation method of the plant composition with tightening and anti-aging effects of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 12 parts of peach resin extract, 5 parts of rhizobium glue extract, 10 parts of gum arabic extract, 17 parts of water lily extract, 10 parts of rosemary herb extract, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 22

The preparation method of the plant composition with tightening and anti-aging effects of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 13 parts of peach resin extract, 6 parts of rhizobium glue extract, 11 parts of gum arabic extract, 13 parts of butterflybush flower extract, 11 parts of rosemary herb extract, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Comparative example 23

The preparation method of the plant composition with the functions of tightening and resisting aging of the embodiment is basically the same as that of the embodiment 1, and the difference is that the weight ratio of each component in the plant composition is as follows: 11.6 parts of peach resin extract, 4.6 parts of rhizobium glue extract, 9.6 parts of gum arabic extract, 11.6 parts of buddleja officinalis extract, 16.6 parts of water lily extract, 45.7 parts of butanediol, 0.15 part of chlorphenesin and 0.15 part of methylparaben.

Effect test

Experiment 1 measurement and calculation of Total alkaloid in plant composition

Centrifuging the obtained plant composition samples (example 1, example 4-8, comparative example 1-11) respectively in a centrifuge at 10000r/min for 30min, collecting supernatant, adding deionized water with the same volume, stirring well, extracting with dichloromethane for 3 times, removing solvent with a rotary evaporator to obtain extract, adding chloroform solvent and fixing volume to 5ml, measuring absorbance value of corresponding plant composition with infrared and calculating to obtain total alkaloid yield of plant composition, wherein the measurement result is shown in figure 1.

As shown in the data in FIG. 1, the content of total alkaloids in the plant composition prepared in example 1 is superior to that in other examples 4-8, which is the best extraction scheme. Compared with comparative examples 3 and 4, the EDSs are superior to the traditional extraction solvent ethanol, and the content of total alkaloids is higher than that of the traditional solvent by more than 21%; compared with the comparative example 1, the method proves that the re-fermentation of the plant filter residue is beneficial to improving the extraction rate of the alkaloid, and is about 11 percent higher; compared with the comparative example 2, the lysozyme can prove that the content of the total alkaloids is effectively increased; comparison with comparative examples 5-11 shows that the mole ratio and water content of EDSs in example 1 are the highest in extraction efficiency of alkaloids.

Experiment 2 measurement of Total sugar in plant composition

Weighing 0.25g of plant composition sample (example 1, example 4-8, comparative example 1-11), respectively adding into a conical flask, adding 50ml of water and 15ml of concentrated hydrochloric acid, carrying out water bath condensation reflux hydrolysis at 100 ℃ for 3h, cooling to room temperature, filtering to remove filter residue, and fixing the volume of the obtained filtrate to 250ml to obtain sample solution. 0.2ml of the sample solution was aspirated into a 10ml stoppered test tube, the volume was made up to 1.0ml with distilled water, 1.0ml of a 5.0% phenol solution was added to the sample solution, 5.0ml of concentrated sulfuric acid (vertical addition) was then added rapidly, and the reaction solution was allowed to stand for 10min. The reaction solution was mixed using a vortex shaker, and then the tube was placed in a 30 ℃ water bath to react for 20min. Taking a proper amount of reaction liquid to test the absorbance at 490nm, calculating the content of total sugar in the plant composition by using a glucose standard curve, and the calculation result is shown in figure 2.

As can be seen from the experimental data in fig. 2, example 1 has an overall sugar content superior to the other examples 4-8, which is the optimal extraction protocol. Compared to comparative example 1, example 1 has a much higher total sugar content, about 40% higher, than comparative example 1, indicating that the remaining plant components can be partially converted to polysaccharides after fermentation by bifidobacteria, greatly increasing the biological total sugar content. Compared with comparative examples 3 and 4, the total sugar content of example 1 and examples 4-8 is higher than that of comparative examples, which shows that the EDSs solvent is favorable for the extraction of biological total sugar, and the total sugar content of example 1 is about 22 percent higher; comparison with comparative examples 5-11 shows that the mole ratio of EDS solvent, water content, and total sugar extraction efficiency in example 1 are the highest.

Experiment 3 measurement of Total phenols in plant compositions

Weighing 11mg of plant groupSamples of the compound (example 1, examples 4 to 8, comparative examples 1 to 11) were each put in a 25mL volumetric flask, dissolved with 30% methanol and diluted to the mark, and 4mL of the solution was precisely sucked up and put in a 25mL volumetric flask, diluted with 30% methanol to the mark. Placing 1mL of the above sample solution in 10mL test tube with plug, processing in dark, adding 20% Folin phenol solution 5mL, shaking, reacting for 5min, adding 5mL of 7.5% Na ₂ CO ₃ Shaking the solution evenly, standing for 1h, and measuring the absorbance at 765 nm. The solution to be measured with the same concentration is repeatedly measured for 3 times respectively to obtain the average value, the total phenol content in the plant composition is calculated by using a polyphenol standard curve, and the measurement and calculation results are shown in figure 3.

From the experimental results of fig. 3, the total phenol content of the plant composition prepared in example 1 is superior to that of the other examples 4 to 8, which is the optimal extraction scheme. Compared with the comparative example 1, the influence of the secondary fermentation on the extraction of polyphenol is smaller than that of polysaccharide and alkaloid (see experiments 1 and 2), but the content of polyphenol is higher than that of polyphenol obtained by the traditional extraction mode (comparative examples 3 and 4), and the EDSs in the invention are favorable for extracting polyphenol.

Experiment 4 Effect of plant composition on fibroblast cells of human skin

To evaluate the effect of the plant composition on fibroblasts of human skin, the proliferation effect of fibroblasts of the plant composition samples (examples 1 to 3, comparative examples 1 to 13) was tested. The specific method comprises the following steps: human skin fibroblasts (HDF) were digested to prepare a cell suspension, which was then inoculated into a 96-well plate at 10000 cells per well. Culturing in DMEM medium for 24 hr, changing culture medium, setting blank group (PBS buffer solution and no light), control group (PBS buffer solution) and experimental group (examples 1-3 and comparative examples 1-13, respectively, and the amount of the added solution is 1% of the cell culture medium), culturing for 24 hr, and providing UVA light (light dose is 5-6J/cm) ² ) And culturing for 24h after the illumination is finished. And adding 10 mu L of sterile CCK-8 solution into each well, continuously culturing for 2h in a dark place, and detecting the absorbance value of 450nm by using an enzyme-labeling instrument. The cell proliferation rate was calculated as = (experimental OD value or control OD value)/blank OD value. The results of the cell proliferation rate measurements are shown in Table 1 below. (Note:* Represents P < 0.05 compared with the control group

As can be seen from the experimental data in the following table, the effect of the plant compositions of examples 1 and 3 on the fibroblasts of human skin is obviously better than that of comparative examples 1 to 7, which indicates that the EDSs adopt choline chloride in a molar ratio: the system of glycerol = 1; the average increment rate of the examples 1-3 is about 10 percent higher than that of the comparative examples 8-13, which shows that the absence of one ingredient in the plant composition has obvious influence on the effect, and the plant ingredient selected by the invention is the optimal scheme of the system.

TABLE 1 Effect of plant compositions on fibroblasts of human skin

Experiment 5 DPPH radical scavenging experiment

The supernatant of the plant composition prepared in examples 1 to 3 and comparative examples 1 to 13 was used as a sample to be tested. A blank group (without adding a sample to be tested), a control group (without adding DPPH), and an experimental group (with the sample to be tested and DPPH added, the addition amount of 0.1mM DPPH is 60% of the final volume) are respectively arranged, and are kept standing for 30min in a dark place at room temperature, centrifuged for 5min at 4000rpm, and the light absorption value at 517nm is tested. Wherein: DPPH free radical clearance (%) = (1-experimental OD value-control OD value)/blank OD value) × 100%. The results are shown in table 2 below.

From the experimental data, the plant compositions of examples 1-3 have better radical clearance than the comparative examples 1-13. The radical scavenging rate of the plant composition of example 1 was much higher than that of comparative example 1, indicating that this process step of fermentation can effectively increase the radical scavenging rate of the composition; compared with comparative example 2, the radical clearance of the plant composition obtained by selecting EDSs is higher than that of the traditional ethanol extraction, which shows that the invention has obvious progress; compared with comparative examples 8 to 13, it is demonstrated that the higher the content of active ingredient of the plant composition of the present invention, the better the radical scavenging effect.

TABLE 2 DPPH radical scavenging test results for botanical compositions

Experiment 6 skin tightening effect experiment of plant composition

Based on the above experiments, in order to verify that the plant composition of the present invention has the effect of tightening skin, the preferred embodiment 1 is compared with the commercial peach-containing latex, and the evaluation is performed by using the human body zygomatic bone skin elasticity test method. The experimental method comprises the following steps: the test subjects washed both hands and face, blotted dry epidermal moisture with disposable facial tissue, entered a test laboratory (temperature: 22 ± 2 ℃, ambient humidity 55%) to sit still for 10min, then tested the skin elasticity value R with the skin elasticity test instrument MPA580, recorded the initial value T0, then smeared the cheekbone with 10% of the basic gel of example 1 and pressed gently with fingers, then sit still for 5min to start measuring skin elasticity change, test time: skin elasticity values R of 5min, 10min, 30min, 1h, 2h, 3h, 6h, and Δ R = R-T0 were calculated, and the results are shown in fig. 4.

Compared with the prior art, the plant composition has the advantages that the skin refinement effect is better than that of the commercially available peach-containing aqueous emulsion, the skin tightening lasting effect reaches the peak at 2h, the attenuation time is relatively slow, and the excellent instant tightening effect is shown.

Experiment 7 application experiment of plant composition in firming essence milk

TABLE 3 formula of firming essence cream

The firming essence milk is prepared according to the formula, and the specific process flow is as follows:

sequentially adding phase A into a beaker, heating to 85 ℃, and stirring and mixing uniformly; sequentially adding the raw materials of the phase B into another beaker, heating to 85 ℃ to dissolve, and stirring and mixing uniformly; and then mixing the phase A and the phase B at 85 ℃, homogenizing by using a homogenizer until the phase A and the phase B are uniformly dispersed, then cooling to 80 ℃, adding the phase C, homogenizing and dispersing, defoaming, cooling to 35 ℃, adding the phase D and the phase E, and uniformly stirring to obtain the firming essence milk. Wherein, the products prepared in example 1, comparative example 3 and comparative example 18 and hyaluronic acid are respectively selected as phase D, and 1%, 3%, 5%, 10% and 20% of the above-mentioned compact essence milk are respectively added to prepare test samples (Table 4).

TABLE 4. Subjects have 5 areas of the forearm smeared with test samples at different addition levels

The product is evaluated for the tightening effect, the skin moisturizing effect is used as an evaluation index, and hyaluronic acid is used as an active substance control. The moisturizing effect evaluation experiment method comprises the following steps: selecting 6 young women of 25-30 years old, dividing forearm of the subject into 5 areas on average under a constant temperature and humidity laboratory environment, smearing each test sample, measuring moisture change value of each area along with time, wherein the total measurement time is 6h, and the test result is shown in figure 5-9.

From the results of the test, it can be seen that when the composition is added in an amount of 5%, the moisture retention level is significantly better than that of hyaluronic acid: the increase in moisture was better than 1% hyaluronic acid over time. Example 1 is the preferred embodiment, compared to the comparative example, in which the moisture retention level is the highest. The composition of the invention has obvious moisturizing effect and obvious improvement.

The above examples are only intended to illustrate the detailed process of the present invention, and the present invention is not limited to the above detailed process, that is, it is not intended that the present invention necessarily depends on the above detailed process to be carried out. It is understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention and the addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.

Claims

1. A preparation method of a plant composition with tightening and anti-aging functions is characterized by comprising the following steps:

s40, mixing plant extract A corresponding to each plant with fermentation filtrate B, adding a preservative to respectively prepare peach resin extract, rhizobia gum extract, gum arabic extract, buddleja officinalis extract, water lily extract and rosemary herb extract, mixing peach resin extract, rhizobia gum extract, gum arabic extract, buddleja officinalis extract, water lily extract and rosemary herb extract, and adding a solvent and a preservative to prepare the plant composition.

2. The method for preparing plant composition with tightening and anti-aging effects as claimed in claim 1, wherein the step S10 is specifically: drying fresh plants, respectively crushing at-35 to 0 ℃, and then sieving with a 200-400-mesh sieve to obtain plant powder, wherein the determination indexes of microorganisms in the plant powder are as follows: the total number of colonies is less than or equal to 500CFU/g, the total number of mould and microzyme is less than or equal to 100CFU/g, and heat-resistant coliform group bacteria, staphylococcus aureus and pseudomonas aeruginosa can not be detected; the determination indexes of the heavy metals are as follows: mercury is less than 1 mg/kg, lead is less than 10 mg/kg, arsenic is less than 2 mg/kg, cadmium is less than 5 mg/kg; pesticide residue could not be detected.

3. The method for preparing the plant composition with the effects of tightening and resisting aging according to claim 1, wherein the plant composition comprises the following components in parts by weight:

5-15 parts of peach resin extract;

2-5 parts of rhizobium glue extracting solution;

3-15 parts of Arabic gum extracting solution;

5-15 parts of a buddleja officinalis extracting solution;

5-18 parts of water lily extracting solution;

5-15 parts of rosemary herb extracting solution;

20-70 parts of butanediol;

0.2-0.4 part of preservative.

4. The method for preparing the plant composition with the effects of tightening and resisting aging according to claim 1, wherein the plant composition comprises the following components in parts by weight:

10 parts of peach resin extracting solution;

3 parts of rhizobium glue extracting solution;

8 parts of Arabic gum extracting solution;

10 parts of a buddleja officinalis extract;

15 parts of water lily extracting solution;

8 parts of rosemary herb extract;

45.7 parts of butanediol;

0.3 part of preservative.

5. The method for preparing the plant composition with tightening and anti-aging effects as claimed in claim 1, wherein the step S20 is specifically: soaking qualified plant powder in purified water according to the weight ratio of 1 (1 to 10), pre-extracting for 1 hour at the temperature of 20 to 45 ℃ at the rotating speed of 50 to 100r/min, then adding a green eutectic solvent with the water ratio of 20% for continuous extraction, wherein the weight ratio of the plant powder to the green eutectic solvent is 1 (1 to 10), and filtering to obtain a filtrate and filter residue after the extraction is finished.

6. The method for preparing plant compositions with firming and anti-ageing effects according to claim 1, wherein said green eutectic solvent consists of HBA and HBD in a molar ratio of 1;

wherein the HBA is lactic acid, and the HBD is any one of sodium acetate, ammonium acetate, glycine and DL-malic acid; or the HBA is choline chloride, and the HBD is any one of glycol, glycerol, oxalic acid, urea, D-glucose, sucrose, lactic acid and acetamide;

the preparation method of the green eutectic solvent comprises the following steps: HBA and HBD are stirred uniformly at 80 ℃ according to the molar ratio, and then 20 percent of water is added and mixed to prepare the green eutectic solvent.

7. The method for preparing plant compositions with tightening and anti-aging effects as claimed in claim 1, wherein step S30 comprises the following steps:

s30b, mixing the filter residue with water according to a volume ratio of 1 to 10, and uniformly stirring at a speed of 100-400r/min to obtain a mixed solution;

s30d, adding lysozyme with the mildew activity of 20000U/mg into the fermentation liquor for breaking the strains, wherein the using amount of the lysozyme is 0.1 to 0.3 percent of the fermentation liquor;

8. The method for preparing the plant composition with tightening and anti-aging effects as claimed in claim 1, wherein in step S40, the preservative is selected from one or more of chlorphenesin, methylparaben, phenoxyethanol, menthone, and hexylene glycol; the solvent is one or more selected from butanediol, propylene glycol, and 1, 3-propylene glycol.

9. A plant composition having firming and anti-aging effects, which is prepared by the method of any one of claims 1 to 8.

10. Use of the plant composition with firming and anti-ageing properties prepared by the preparation method according to any one of claims 1 to 8 in cosmetics.