CN117064824A - Method for preparing cosmetic raw material by using distilled spirit vinasse extract and application thereof - Google Patents
Method for preparing cosmetic raw material by using distilled spirit vinasse extract and application thereof Download PDFInfo
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- CN117064824A CN117064824A CN202310629349.0A CN202310629349A CN117064824A CN 117064824 A CN117064824 A CN 117064824A CN 202310629349 A CN202310629349 A CN 202310629349A CN 117064824 A CN117064824 A CN 117064824A
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- raw material
- cosmetic raw
- cosmetic
- distilled
- distilled spirit
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- 229960003512 nicotinic acid Drugs 0.000 description 1
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- FDRQPMVGJOQVTL-UHFFFAOYSA-N quercetin rutinoside Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 FDRQPMVGJOQVTL-UHFFFAOYSA-N 0.000 description 1
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- IKGXIBQEEMLURG-BKUODXTLSA-N rutin Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](C)O[C@@H]1OC[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](OC=2C(C3=C(O)C=C(O)C=C3OC=2C=2C=C(O)C(O)=CC=2)=O)O1 IKGXIBQEEMLURG-BKUODXTLSA-N 0.000 description 1
- ALABRVAAKCSLSC-UHFFFAOYSA-N rutin Natural products CC1OC(OCC2OC(O)C(O)C(O)C2O)C(O)C(O)C1OC3=C(Oc4cc(O)cc(O)c4C3=O)c5ccc(O)c(O)c5 ALABRVAAKCSLSC-UHFFFAOYSA-N 0.000 description 1
- 235000005493 rutin Nutrition 0.000 description 1
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- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
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- 238000003860 storage Methods 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-N sulfuric acid Substances OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 238000004704 ultra performance liquid chromatography Methods 0.000 description 1
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- 239000011579 vitamin B4 Substances 0.000 description 1
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Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
- A61K8/97—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution from algae, fungi, lichens or plants; from derivatives thereof
- A61K8/9783—Angiosperms [Magnoliophyta]
- A61K8/9794—Liliopsida [monocotyledons]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/02—Preparations for care of the skin for chemically bleaching or whitening the skin
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q19/00—Preparations for care of the skin
- A61Q19/08—Anti-ageing preparations
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/805—Corresponding aspects not provided for by any of codes A61K2800/81 - A61K2800/95
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
- A61K2800/80—Process related aspects concerning the preparation of the cosmetic composition or the storage or application thereof
- A61K2800/85—Products or compounds obtained by fermentation, e.g. yoghurt, beer, wine
Abstract
The invention discloses a method for preparing a cosmetic raw material by using a distilled spirit vinasse extract, which comprises the following steps: (1) semisolid enzyme extraction; (2) solid-liquid separation; (3) membrane separation; (4) sterilization and corrosion prevention: sterilizing L3, namely sterilizing distilled spirit vinasse extract, and adding a preservative after sterilization to obtain the raw material of the cosmetics. The cosmetic raw material is used for repairing cell injury. The material extracted by the method has good quality, can be used as a cosmetic raw material, and can improve the utilization rate of vinasse and obtain a high-activity cosmetic raw material; can also be applied to medicines or skin care products for repairing cell injury.
Description
Technical Field
The invention relates to the technical field of cosmetic raw materials, in particular to a method for preparing a cosmetic raw material by using distilled spirit vinasse extract and application thereof.
Background
In recent years, consumer demand for cosmetics that are green, safe and efficacious is increasing, and the core competitiveness of the cosmetic industry is gradually condensing in both safety and efficacy. Therefore, safe cosmetic raw materials and products with outstanding efficacy have good prospects in the market.
Grain such as sorghum, barley, wheat and corn contains rich nutrients including protein, beta-glucan, phenolic substances, flavonoid substances, cellulose and the like, and is a common raw material for brewing wine. The distillers 'grains are by-products of the fermentation of the grains by the yeast, and the residual active metabolites of the distilled liquor are not utilized in the distillers' grains. The distilled spirit vinasse is rich in various healthy and natural fermentation source skin beautifying components, and mainly contains amino acids in natural moisturizing factors, so that human cells can be synthesized, degradation or aging of the cells can be prevented, bone collagen can be decomposed, and skin can keep elasticity. In addition, the moisture-preserving components such as polysaccharide, organic acid and polyphenol in the distilled grains are beneficial to improving the skin moisture-preserving function, moistening the skin and improving the skin problems caused by skin dryness. The distilled spirit vinasse has the advantages of wide source, high yield and low price, has great development potential and good application prospect, and the main treatment mode is to discard or manufacture animal feed, so that the environment is influenced, and the value of the distilled spirit vinasse is not fully utilized. At present, a development and utilization way of distillers' grains with high economic added value is still lacking.
The Chinese patent application CN110179932A discloses a preparation method of a small molecular peptide of vinasse and application of the small molecular peptide of vinasse in skin care products, wherein the vinasse is subjected to deep processing by adopting a specific vacuum puffing technology, vinasse protein is extracted by alkali dissolution and acid precipitation, and simultaneously, a protease is utilized to hydrolyze a vinasse protein extract, so that the small molecular peptide is used for preparing moisturizing repair emulsion. However, this method has the following drawbacks: (1) The alkali-dissolution alcohol precipitation method for extracting protein is easy to generate compound structural change under severe conditions, and has harm to equipment and environment; (2) The enzyme preparation has harsh protein hydrolysis reaction conditions, the enzyme activity is regulated and controlled by a plurality of factors such as temperature, pH, substrate concentration and the like, and the enzyme activity is considered to be maintained in actual production.
At present, the extraction method of active substances in distilled spirit vinasse mainly comprises an acid-base extraction method and a high-temperature extraction method. The acid-base extraction method is widely used in extracting active substances, has the characteristics of low degradation cost and mature technology, but the reaction process is severe, the structure of the active substances can be damaged, equipment is easy to corrode, and the environment is polluted to a certain extent. High temperature extraction requires heating to the required temperature and maintaining the temperature for a proper time, but the heating temperature and time must be strictly controlled in the operation process, the reflux temperature is kept stable, and the structure of the active substances in the reflux temperature can be damaged due to overhigh temperature or overlong time. In the process of extracting active substances from the fermentation product, the composition of substances in the fermentation liquid is complex, the filtration is difficult, and the fermentation liquid after solid-liquid separation has poor clarity, high impurity content and weak stability, so that the sedimentation or layering problem often occurs in the storage process.
Disclosure of Invention
In order to solve the technical problems, the invention provides a method for preparing a cosmetic raw material by using a distilled grain extract of white spirit and application thereof, and solves the technical problems of poor clarity, high impurity content and weak stability of distilled grain in the prior art.
The invention obtains the cosmetic active raw material which is clear, has high content of active ingredients and stable state by extracting the nutritional ingredients and the active substances of the distillers' grains, can prevent sedimentation or layering phenomenon of the product in the use process, ensures the efficacy and quality of the product, realizes the specific, efficient and mild preparation of the cosmetic active raw material with outstanding efficacy, and is applied to industrialization.
In order to achieve the above purpose, the technical scheme provided by the invention is as follows:
a method for preparing cosmetic raw materials by using distilled spirit vinasse extract comprises the following operation steps:
(1) Semi-solid enzymatic extraction: cooling distilled grains of white spirit, adding mixed enzyme for enzymolysis extraction to obtain semi-solid enzymolysis product L1, wherein the mixed enzyme is at least two of cell lyase, protease or cellulase;
(2) Solid-liquid separation: performing solid-liquid separation on the semi-solid enzymolysis product L1 to obtain clear liquid L2;
(3) Membrane separation: performing secondary physical membrane separation on the clear liquid L2 to obtain a small molecular active concentrated liquid L3 with the molecular weight smaller than 1 ten thousand Da, namely a distilled grain extract of the distilled spirit; the secondary physical membrane separation is a high-efficiency green membrane separation technology for connecting microfiltration and ultrafiltration in series, and the substances to be separated are sequentially separated by the secondary physical membrane; the secondary physical membrane separation is ceramic membrane-ultrafiltration separation or metal membrane-ultrafiltration separation;
(4) Sterilization and corrosion prevention: sterilizing L3, namely sterilizing distilled spirit vinasse extract, and adding a preservative after sterilization to obtain the raw material of the cosmetics.
Preferably, the extraction temperature in the step (1) is 25 ℃ and the extraction time is 2 hours; the separation method in step (2) comprises at least one of pressing, gravity settling, centrifugal separation or filtration; the microfiltration in the step (3) is a ceramic membrane or a metal membrane.
Preferably, the sterilization mode in the step (4) is one of normal temperature steam sterilization, low temperature pasteurization or microwave sterilization.
Preferably, the preservative in the step (4) is one of phenoxyethanol, benzyl alcohol, benzoic acid or methylparaben.
The cosmetic raw material contains substances such as caffeic acid, proline oxide, muscone, azalea glycoside, xylitol and lactose, etc., and is helpful for preventing skin injury and melanoid deposition, and has antioxidant, skin whitening and moisturizing effects.
The cosmetic raw material prepared by the method is applied to repairing cell injury.
The cosmetic raw material prepared by the method is applied to the preparation of medicines for repairing cell injury.
The cosmetic raw material prepared by the method is applied to skin care.
The cosmetic raw material prepared by the method is applied to skin care after being prepared into distilled spirit vinasse cosmetic water.
Preferably, the distilled spirit vinasse cosmetic water is prepared according to the following method:
weighing the following raw materials in percentage by weight: 2% of glycerin, 5% of butanediol, 0.01% of xanthan gum, 0.05% of EDTA disodium, 0.3% of hyaluronic acid, 20% of the raw materials of the cosmetics, 0.01% of citric acid, 0.03% of sodium citrate and the balance of deionized water; preparing a water phase pot: heating and uniformly mixing deionized water and cosmetic raw materials according to the formula amount, and stirring to obtain a water phase pot;
(II) weighing butanediol and xanthan gum according to the formula, stirring uniformly in advance, and adding into a water phase pot;
(III) weighing the glycerol, EDTA disodium and hyaluronic acid according to the formula amount, adding the mixture into a water phase pot, stirring uniformly to dissolve powder substances in the liquid, preserving heat, and cooling in a cold water bath;
and (IV) adding the citric acid and the sodium citrate in the formula amount, stirring, dissolving, uniformly mixing, and homogenizing to obtain the distilled grain cosmetic water of the white spirit.
Compared with the prior art, the invention has the beneficial effects that:
the semi-solid enzymatic extraction process adopted by the method has the advantages of simple operation, environmental protection, high preparation efficiency and low energy consumption, and the mixed enzymolysis technology can target the mycoprotein, undegraded cellulose and the like remained in the enzymolysis vinasse and further promote the dissolution of intracellular active substances; simultaneously, the distillers' grains extract prepared by the secondary membrane separation technology has stable quality, high clarity and strong activity; furthermore, the extracted substances have good quality, can be used as cosmetic raw materials, can improve the utilization rate of vinasse, and can obtain high-activity cosmetic raw materials; can also be applied to medicines or skin care products for repairing cell injury.
Brief description of the drawings
FIG. 1 is a multiple of the difference between the small molecule substances in example 1 and comparative example 1 (example 1/comparative example 1).
FIG. 2 is a fold difference between partial amino acids and dipeptides in example 1 and comparative example 1 (example 1/comparative example 1).
Fig. 3 is the effect of different concentrations of cosmetic raw material on L929 cell proliferation, p <0.001 compared to control.
FIG. 4 shows different concentrations of H 2 O 2 Effect on cell viability, p compared to control group<0.01，***p<0.001。
Fig. 5 is a graph showing the repair effect of 25% cosmetic material on oxidatively damaged cells, with p <0.01 and p <0.001 compared to model group.
Fig. 6 is a diagram of sensory evaluation radar of distilled spirit vinasse cosmetic water prepared by application example.
Detailed Description
The following detailed description, in conjunction with the accompanying drawings, describes in detail, but it is to be understood that the scope of the invention is not limited to the specific embodiments. The raw materials and reagents used in the examples were commercially available unless otherwise specified.
In the examples, the culture medium used for detecting the activity of the cells subjected to the repair of the oxidative damage was DMEM medium (containing 10% fetal calf serum and 1% penicillin-streptomycin solution), and the cells were sterilized at 121℃for 20 minutes.
Example 1
A method for preparing cosmetic raw material by using distilled spirit vinasse extract comprises the following operation steps:
(1) Semi-solid enzymatic extraction: cooling distilled grains of distilled liquor, taking 1kg, adding deionized water, and the feed liquid ratio is 2:1 (mass: volume), adding mixed enzyme for enzymolysis extraction at 25deg.C for 2 hr to obtain semisolid enzymolysis product L1, wherein the mixed enzyme is protease and cellulase (mixed at any ratio);
(2) Solid-liquid separation: performing solid-liquid separation on the semi-solid enzymolysis product L1 by squeezing to obtain clear liquid L2;
(3) Membrane separation: the clear liquid L2 is subjected to secondary physical membrane separation to remove macromolecular impurities in the extracting solution, wherein the secondary physical membrane separation is an efficient green membrane separation technology for connecting microfiltration and ultrafiltration in series, the secondary physical membrane separation is specifically ceramic membrane-ultrafiltration separation, ultrafiltration membrane filtration clarification with the molecular weight cutoff of 10k Da is adopted, suspended matters, particles, bacteria and the like are separated, so as to purify and concentrate vinasse extracting solution to obtain small molecular active concentrate L3 with the molecular weight less than 1 ten thousand Da, namely the vinasse extract of white wine.
(4) Sterilization and corrosion prevention: and (3) performing low-temperature pasteurization on L3, namely sterilizing distilled spirit vinasse extract, adding 0.5% phenoxyethanol as a preservative after sterilization, thus obtaining a cosmetic raw material, and storing the cosmetic raw material at 4 ℃.
Example 2
A method for preparing cosmetic raw material by using distilled spirit vinasse extract comprises the following operation steps:
(1) Semi-solid enzymatic extraction: cooling distilled grains of distilled liquor, taking 1kg, adding deionized water, and the feed liquid ratio is 2:1 (mass: volume), adding mixed enzyme for enzymolysis extraction at 25deg.C for 2 hr to obtain semisolid enzymolysis product L1, wherein the mixed enzyme is cell lyase and protease (mixed at any ratio);
(2) Solid-liquid separation: performing solid-liquid separation on the semi-solid enzymolysis product L1, wherein the separation method comprises squeezing and centrifugal separation to obtain clear liquid L2;
(3) Membrane separation: performing secondary physical membrane separation on clear liquid L2 to remove macromolecular impurities in the extract, wherein the secondary physical membrane separation is an efficient green membrane separation technology for connecting microfiltration and ultrafiltration in series, the secondary physical membrane separation is specifically ceramic membrane-ultrafiltration separation, ultrafiltration membrane filtration clarification with a molecular weight cutoff of 10k Da is adopted to separate suspended matters, particles, bacteria and the like, so as to purify and concentrate vinasse extract to obtain small molecular active concentrate L3 with a molecular weight of less than 1 ten thousand Da, namely the vinasse extract of white wine;
(4) Sterilization and corrosion prevention: sterilizing L3, i.e. distilled spirit vinasse extract, by low temperature pasteurization, adding 0.5% methyl p-hydroxybenzoate as antiseptic after sterilization to obtain cosmetic raw material, and storing at 4deg.C.
Comparative example 1
A method for preparing cosmetic raw material by using distilled spirit vinasse extract comprises the following specific steps:
(1) Weighing 1kg of dried distillers' grains, adding 50% ethanol in volume concentration, and mixing the materials according to a feed liquid ratio of 1:10 (mass: volume) at 25deg.C for 2 hr to obtain suspension L1;
(2) Filtering the suspension L1 to intercept insoluble substances, centrifuging to obtain filtrate, and collecting supernatant to obtain supernatant L2;
(3) Concentrating the supernatant L2 at 55 ℃ under reduced pressure until the ethanol content in the supernatant is 0, and then removing impurities by suction to obtain L3;
(4) Concentrating and suction-filtering the liquid L3 in the step (3), heating at 80 ℃ for 20min for sterilization, adding 0.5% phenoxyethanol as a preservative to prepare a cosmetic active raw material, and storing in the temperature of 4 ℃.
Detection of
The ingredients and the free radical scavenging ability of the cosmetic raw materials of example 1 and comparative example 1 were measured according to the following method, and the results are shown in Table 1, with the commercial products SK-II essence and SK-II skin-tendering lotion as controls; the differential small molecule substances of the cosmetic raw materials of example 1 and comparative example 1 were identified according to the following method, and the results are shown in fig. 1 and 2; the cosmetic raw material prepared in example 1 was tested for its effect of repairing oxidatively damaged cells according to the following method, and the results are shown in FIGS. 3 to 5.
(1) Component analysis
(1) Protein content
1mL of the cosmetic raw material of example 1, the cosmetic raw material of comparative example 1, the SK-II essence and the SK-II skin care lotion which are control products are taken respectively and put in test tubes, 4mL of biuret reagent is added respectively, after shaking evenly, the mixture is placed for 30 minutes at room temperature (25 ℃), and the absorbance at the wavelength of 540nm is measured. And (5) establishing a standard curve by taking casein as a standard substance, and calculating the protein content.
(2) Reducing sugar content
The cosmetic raw material of the example 1, the cosmetic raw material of the comparative example 1, the SK-II essence and the SK-II skin tendering lotion which are control products are diluted by adding water until the content of reducing sugar is in the range of 0-0.1mg/mL, 1.0mL of diluted sample solution is taken and put in a test tube, 2.0mL of DNS reagent is added, boiling water is boiled for 5min, water is used for supplementing to 15mL after cooling, and absorbance is measured at the wavelength of 540 nm. And (3) establishing a standard curve by taking glucose as a standard substance, and calculating the reducing sugar content.
(3) Total sugar content
The cosmetic raw material of example 1, the cosmetic raw material of comparative example 1, the SK-II essence and the SK-II skin tendering lotion which are control products are diluted by adding water until the total sugar content is in the range of 0-0.1mg/mL, respectively taking 1mL of diluted sample into a test tube, adding 1mL of 5% phenol solution and 5mL of concentrated sulfuric acid, shaking uniformly, cooling to room temperature, and measuring the absorbance at 490nm wavelength. And (5) establishing a standard curve by taking glucose as a standard substance, and calculating the total sugar content.
(4) Total phenol content
Diluting the cosmetic raw material of example 1, the cosmetic raw material of comparative example 1, the SK-II essence and the SK-II skin care lotion of the control products respectively until the total flavone content is in the range of 0-1.0mg/mL, respectively taking 1mL of diluted sample into a test tube, adding 3mL of Fu Lin Fen reagent, uniformly mixing, standing for 5min, adding 6mL of 7.5% Na 2 CO 3 The solution was mixed well and then fixed to 25mL, shaken well, left in the dark for 2h, and the absorbance was measured at 760 nm. And (3) establishing a standard curve by taking gallic acid as a standard substance, and calculating the total phenol content.
(5) Total flavone content
The cosmetic raw material of example 1, the cosmetic raw material of comparative example 1, the SK-II essence and the SK-II skin tendering lotion of the control products are respectively diluted until the total flavone content is in the range of 0-0.2mg/mL, 1mL of diluted sample is respectively taken in a test tube, 1mL of 10% aluminum nitrate solution and 1mL of 5% sodium acetate solution are added, the volume is fixed to 10mL by 95% ethanol, shaking is carried out, standing is carried out for 30min, and the absorbance is measured at 415 nm. And (3) establishing a standard curve by taking rutin as a standard substance, and calculating the total flavone content.
(2) Free radical scavenging ability
(1) DPPH radical scavenging rate
100. Mu.L of the cosmetic raw material of example 1, the cosmetic raw material of comparative example 1, the SK-II essence and SK-II skin tendering lotion which are control products are respectively taken and mixed with 500. Mu.L of ethanol, 125. Mu.L of DPPH ethanol solution (2% w/v) is added, the mixture is uniformly mixed and then reacted for 60 minutes in a dark place, and the absorbance at 517nm is measured. The DPPH radical scavenging rate of the sample was calculated by the following formula, with the same volume of water as the control instead of the sample:
DPPH radical clearance (%) = (control OD-experimental OD)/control OD x 100%
(2) Free radical scavenging rate of active oxygen
0.6mL of phenanthroline (5 mmol/L) was added to 0.4mL of phosphate buffer (0.2M, pH 7.4) and mixed, and then 0.6mL of cosmetic material of example 1 and comparative sample were addedExample 1 cosmetic raw materials and control products SK-II essence and SK-II skin tendering lotion and 0.6mL EDTA (15 mmol/L) were mixed again and then 0.6mL FeSO was added 4 The solution (5 mmol/L) was made up to a volume of 2.8mL with deionized water, and after thorough mixing, 0.8mL of H was added 2 O 2 (0.1%) was incubated at 37℃for 1h after shaking, and the absorbance at 536nm was measured. The same volume of water is used as a damage group instead of a sample, and deionized water is used as a substitute for H 2 O 2 As an undamaged group, the OH clearance of the samples was calculated by the following formula:
OH clearance (%) = (experimental group OD value-damaged group OD value)/(undamaged group OD value-damaged group OD value) ×100%
(3) Identification of differential Small molecule substances
The cosmetic raw materials prepared in example 1 and comparative example 1 were extracted twice with 6-fold volume of methanol, respectively, mixed for 1 minute, centrifuged at 13000 Xg for 15 minutes, and filtered with a 0.22 μm filter membrane, and then analyzed by high performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS). High performance liquid chromatography was equipped with ACQUITY UPLC BEH C column at 30 ℃. The gradient mobile phase consisted of 0.1% formic acid water (a) and acetonitrile (B). The gradient conditions of the mobile phase are as follows: 5% of B lasted 2 minutes, then B increased from 5% to 95% over 14 minutes, followed by 5% of B isocratic elution for 2 minutes. The gradient duration was 18 minutes and the flow rate was 0.30mL/min. Electrospray ionization (ESI) source interfaces were used, operating in positive and negative modes. The resolution of the primary mass spectrum was 70,000, the target for automatic gain control was 3e6, the maximum IT was 50ms, and the dynamic exclusion time was 10.0s. The resolution of the secondary mass spectrum was 17,500 and the normalized collision energy was 30eV. The voltages for the positive mode (ESI+) and the negative mode (ESI-) were set at 30 and 30V, with mass spectra acquisition ranging from 100 to 1000m/z.
(4) Repair of oxidative damage cell Activity
(1) Effect on L929 cell proliferation
Selecting L929 fibroblast in logarithmic growth phase, and adjusting cell concentration to 5.0X10/ml after digestion 5 Inoculating 100 μl of each well into 96-well plate at 37deg.C with 5% CO 2 Culturing in an incubator for 24 hours. Discarding the old medium, culturing in serum-free medium for 24 hr, and sucking out into 96-well plateThe experimental groups were added with the culture medium containing different concentrations of the cosmetic raw materials (5, 10, 15, 25, 50%, v/v) prepared in example 1, the control group was the culture medium without the sample solution, and the blank group was the pure culture medium. After 24h incubation, 10. Mu.L MTT reagent was added and incubation was continued for 4h, 200. Mu.L DMSO was added and shaking at low speed for 5min to determine absorbance at 490 nm. Cell viability was calculated according to the following formula:
cell viability (%) = (experimental OD value-blank OD value)/(control OD value-blank OD value) ×100%
(2) For H 2 O 2 Repair of induced L929 cell damage
Establishment of cell damage model: selecting L929 fibroblast in logarithmic growth phase, and adjusting cell concentration to 5.0X10/ml after digestion 5 Inoculating 100 μl of each well into 96-well plate at 37deg.C with 5% CO 2 Culturing in an incubator for 24 hours. Discarding the old medium, culturing in serum-free medium for 24 hr, sucking out the medium in 96-well plate, and adding H with different concentration 2 O 2 (0.98, 1.47, 1.86, 2.94, 4.41 mmol/L) of culture medium, control group was H-free 2 O 2 Is pure medium. After 24h incubation, 10. Mu.L MTT reagent was added, incubation was continued for 4h, 200. Mu.L DMSO was added, shaking at low speed for 5min, absorbance at 490nm was measured, and cell viability was calculated.
Cosmetic raw material pair H 2 O 2 Inducing repair of L929 cell damage: selecting L929 fibroblast in logarithmic growth phase, and adjusting cell concentration to 5.0X10/ml after digestion 5 Inoculating 100 μl of each well into 96-well plate at 37deg.C with 5% CO 2 Culturing in an incubator for 24 hours. The old medium was discarded, and after culturing in serum-free medium for 24 hours, 2.94mmol/L H was added 2 O 2 After 24h of culture to induce cell damage, the culture medium in the 96-well plate was aspirated, 25% of the cosmetic raw material prepared in example 1 was added to the experimental group (25% was the cosmetic raw material prepared in example 1 diluted with deionized water to 25% by mass concentration), the control group was the culture medium without the cosmetic raw material, and the blank group was the pure culture medium. After 24h of incubation, 10. Mu.was addedL MTT reagent, further culturing for 4h, adding 200 mu L DMSO, shaking at low speed for 5min, measuring absorbance at 490nm, and calculating cell survival rate.
TABLE 1 analysis of ingredients of cosmetic raw materials and commercially available products prepared in example 1 and free radical scavenging ability test
| Example 1 | Comparative example 1 | SK-II essence | SK-II skin care lotion | |
| Protein content/% | 10.851±0.102 | 1.776±0.377 | 0.182±0.014 | 0.016±0.004 |
| Reducing sugar content/% | 0.374±0.001 | 0.565±0.036 | 0.115±0.010 | 0.064±0.006 |
| Total sugar content/% | 3.640±0.136 | 0.894±0.450 | 1.514±0.162 | 0.451±0.041 |
| Total phenol content/% | 0.077±0.006 | 0.046±0.000 | 0.005±0.000 | 0.010±0.000 |
| Total flavone content/% | 0.007±0.000 | 0.005±0.000 | N.D. | 0.001±0.001 |
| DPPH radical scavenging/% | 57.14±5.10 | 78.69±0.81 | 2.01±0.81 | 2.90±0.73 |
| Active oxygen radical scavenging/% | 20.54±1.31 | 85.69±4.19 | 12.88±3.87 | N.D. |
Note that: n.d. indicates undetected.
As can be seen from the data in Table 1, the cosmetic raw material prepared in example 1 has higher protein, total sugar, total phenol and total flavone contents than those of comparative example 1 and the commercially available SK-II products. The protein is a nutrient substance necessary for the growth and development of tissues and cells, and can moisten skin, nourish cells and promote the healthy growth and development of skin tissues. The polysaccharide can control cell division and differentiation, and regulate cell growth and aging recovery. The polyphenol has biological activities of resisting oxidation, scavenging free radicals, resisting bacteria, preserving freshness, etc., and can relieve muscle fatigue and repair muscle injury. The flavone has biological activities such as oxidation resistance, aging resistance and inflammation resistance besides moisturizing effect, and the addition of the flavone component in the cosmetics is helpful for recovering elasticity and luster of dry and loose skin. From the free radical scavenging capability test of the product in example 1 and the commercial product, the cosmetic raw material prepared in the example of the invention has higher DPPH free radical and active oxygen free radical scavenging rate and is obviously better than the commercial SK-II product. Meanwhile, the data in table 1 show that the radical scavenging rate of comparative example 1 is higher than that of example 1, probably because the alcohol-soluble substance in comparative example 1 is more conducive to scavenging DPPH radicals and reactive oxygen radicals.
In example 1, as shown in fig. 1, the cosmetic raw materials in comparative example 1 were detected with 1645 kinds of substances, and in example 1, the content of small molecules is larger than in comparative example 1, and 1414 kinds of substances with a difference multiple of more than 2 are detected, which means that the preparation method of example 1 is more favorable for extracting small molecule substances in distilled spirit vinasse compared with comparative example 1. In particular, the amino acid and dipeptide content in example 1 was significantly higher than that in comparative example 1, and fig. 2 shows the fold difference in partial amino acid and dipeptide. In particular, high levels of vitamins such as pantothenic acid, vitamin B3, vitamin B4 and biotin, and bacteriostatic substances such as undecylglycine, ascopyrone M and actinomycin, and small molecules such as dihydrocaffeic acid, proline oxide, xylitol, lactose, muscone, azalide are found in the cosmetic raw materials of example 1. Wherein, the dihydrocaffeic acid has anti-inflammatory and antioxidant activities, and can prevent skin injury induced by ultraviolet rays; the proline oxide has the inhibition effect on tyrosine oxidase, can prevent the deposition of melanoid and whiten skin; xylitol and lactose can be used as humectant, and have high stability; muscone can significantly reduce inflammatory cytokine levels; azalea glycoside has antioxidant and antiviral activities.
As shown in FIG. 3, compared with the control group, after 5-50% of the raw material of the cosmetic prepared in example 1 is added, the survival rate of L929 cells is significantly increased (p < 0.001), which indicates that the raw material of the cosmetic in example 1 is safe and nontoxic to cells, can be used in the cosmetic field, and has the effects of promoting cell proliferation, activating cells, enhancing cell metabolism and the like.
By H 2 O 2 Treatment, a cell oxidative stress injury model was established to investigate the repair effect of the cosmetic raw material of example 1 on injured cells. As shown in FIG. 4, a different concentration of H was added compared to the control group 2 O 2 After 24h of action, the cell viability decreased. When H is 2 O 2 At a concentration of 1.86mmol/L, cell viability was significantly reduced to 59.24% (p)<0.01 When H) 2 O 2 At a concentration of 2.94mmol/L, the cell viability was significantly reduced to 33.45% (p)<0.001). Subsequent experiments selected 2.94mmol/L H 2 O 2 The effect of the cosmetic material of example 1 on repairing injured cells was examined by inducing oxidative damage of L929 cells. As shown in FIG. 5, the cell viability of the model group was significantly reduced to 60.04% (p)<0.001). After 25% cosmetic material treatment, the cell viability was significantly increased in the experimental group compared to the model group (p<0.01 82.45%, indicating that the cosmetic raw material of example 1 can improve oxidative damage of cells.
Application example
The application example is the preparation of distilled spirit vinasse toning lotion, which comprises the following raw materials in percentage by weight: 2% of glycerin, 5% of butanediol, 0.01% of xanthan gum, 0.05% of EDTA disodium, 0.3% of hyaluronic acid, 20% of raw materials of cosmetics prepared in example 1, 0.01% of citric acid, 0.03% of sodium citrate and the balance of deionized water. The preparation method comprises the following specific steps:
preparing an aqueous phase pot: heating deionized water with the formula amount and the raw materials of the cosmetics prepared in the example 1 to 85 ℃, and uniformly mixing and stirring to obtain a water phase pot;
(II) weighing butanediol and xanthan gum according to the formula, stirring uniformly in advance, and adding into a water phase pot;
(III) weighing the glycerol, EDTA disodium and hyaluronic acid according to the formula amount, adding into a water phase pot, stirring uniformly to dissolve powder substances in the liquid, preserving heat for 10min, and cooling to 45 ℃ in a cold water bath;
and (IV) adding the citric acid and the sodium citrate in the formula amount, stirring, dissolving, uniformly mixing, and homogenizing to obtain the distilled grain cosmetic water of the white spirit.
The distilled spirit vinasse cosmetic water prepared by adopting the application example is tested for use effect, and the results are shown in Table 2 and FIG. 6.
(1) Moisture effect test
20 volunteers with ages of 20-45 years are selected, 10 men and women are respectively, and the ages and sexes are evenly distributed. After the subject cleans both hands with clear water, the initial skin moisture content, elasticity and oil content of the hands were measured using a skin moisture content tester. The subject takes 1mL of distilled spirit vinasse cosmetic water prepared in the application example to carry out the hand back, lightly smears and massages the hand back for about 1min to completely absorb the distilled spirit vinasse cosmetic water, and detects the water content, the elasticity and the oil content of the skin again after 5 min. The test results are shown in Table 2.
TABLE 2 application example preparation of distilled spirit lees Pre-and post-skin Water content results
| Moisture (%) | Elasticity (%) | Oil content (%) | |
| Before use | 23.9±0.1 | 29.6±0.1 | 46.9±0.1 |
| UsingRear part (S) | 56.3±0.1 | 23.3±0.1 | 60.0±0.0 |
The moisture, oil content and elasticity of the skin are important physical parameters that indicate the skin condition. As shown in table 2, after the subject uses the distilled spirit vinasse cosmetic water prepared by the application example of the present invention, the skin moisture is significantly increased, the elasticity is slightly decreased, and the oil content is slightly increased. The raw materials of the cosmetic used in the invention contain various active ingredients such as protein, polysaccharide, polyphenol and the like, and the active ingredients can ensure that the prepared toning lotion has good effects of moisturizing, whitening, anti-inflammatory, anti-aging and the like. The test results show that the distilled spirit vinasse cosmetic water prepared by the application example can effectively supplement moisture for skin, and the increase of oil content is beneficial to forming a layer of protective film on the surface of the skin, reducing the loss of the moisture of the skin, enhancing the moisturizing effect and repairing the skin barrier.
(2) Sensory evaluation
20 volunteers with ages of 20-45 years are selected, 10 men and women are respectively, and the ages and sexes are evenly distributed. Through means of observation, sniffing and the like, the subjects score indexes such as color, state, smell and the like of the distilled spirit vinasse cosmetic water prepared by the application example, and each index is fully scored for 15 minutes. And (3) scoring indexes such as smearing feeling, moisture retention, mildness and the like of the toning lotion prepared by the smearing application of the subject, fully scoring each index for 15 minutes, comprehensively testing the result, and drawing a radar chart. The results are shown in FIG. 6.
As can be seen from fig. 6, most subjects were satisfied with the state and color of the lotion prepared in the application example, and the subjects showed no stickiness, easy absorption, and no irritation with respect to the smearing feeling, the moisture retention, and the mildness of the product, and the smell score of the product was general, because the lotion prepared in the application example was free from the addition of other smell components other than the raw materials of the skin care product, and the addition of the functional component having fragrance was considered in the actual production.
The invention uses the two-stage membrane separation technology combining microfiltration and ultrafiltration, and uses the filter membrane with proper aperture to intercept suspended matters, particles, bacteria and other macromolecular substances so as to purify and concentrate active ingredients in the raw materials of cosmetics, and the obtained raw materials have good quality and strong product stability, no chemical reagent is required to be added in the process, and the equipment can automatically operate, and has the advantages of low energy consumption, energy conservation and environmental protection.
The foregoing descriptions of specific exemplary embodiments of the present invention are presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application to thereby enable one skilled in the art to make and utilize the invention in various exemplary embodiments and with various modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.
Claims (9)
1. A method for preparing a cosmetic raw material by using distilled spirit vinasse extract, which is characterized by comprising the following operation steps:
(1) Semi-solid enzymatic extraction: cooling distilled grains of white spirit, adding mixed enzyme for enzymolysis extraction to obtain semi-solid enzymolysis product L1, wherein the mixed enzyme is at least two of cell lyase, protease or cellulase;
(2) Solid-liquid separation: performing solid-liquid separation on the semi-solid enzymolysis product L1 to obtain clear liquid L2;
(3) Membrane separation: performing secondary physical membrane separation on the clear liquid L2 to obtain a small molecular active concentrated liquid L3 with the molecular weight smaller than 1 ten thousand Da, namely a distilled grain extract of the distilled spirit; the secondary physical membrane separation means that microfiltration and ultrafiltration are connected in series;
(4) Sterilization and corrosion prevention: sterilizing L3, namely sterilizing distilled spirit vinasse extract, and adding a preservative after sterilization to obtain the raw material of the cosmetics.
2. The method for preparing a cosmetic raw material using a distilled spirit vinasse extract according to claim 1, wherein: the extraction temperature in the step (1) is 25 ℃, and the extraction time is 2 hours; the separation method in step (2) comprises at least one of pressing, gravity settling, centrifugal separation or filtration; the microfiltration in the step (3) is a ceramic membrane or a metal membrane.
3. The method for preparing a cosmetic raw material using a distilled spirit vinasse extract according to claim 1, wherein: the sterilization mode in the step (4) is one of normal temperature steam sterilization, low temperature pasteurization or microwave sterilization.
4. The method for preparing a cosmetic raw material using a distilled spirit vinasse extract according to claim 1, wherein: the preservative in the step (4) is one of phenoxyethanol, benzyl alcohol, benzoic acid or methylparaben.
5. Use of the cosmetic raw material obtained by the method according to any one of claims 1 to 4 for repairing cell damage.
6. Use of the cosmetic raw material obtained by the method according to any one of claims 1 to 4 for the preparation of a medicament for repairing cell damage.
7. Use of the cosmetic raw material obtained by the process according to any one of claims 1 to 4 for skin care.
8. Use of the cosmetic raw material obtained by the method according to any one of claims 1-4 in skin care after preparation of distilled spirit vinasse cosmetic water.
9. The use according to claim 8, wherein the distilled spirit vinasse cosmetic water is prepared according to the following method:
weighing the following raw materials in percentage by weight: 2% of glycerin, 5% of butanediol, 0.01% of xanthan gum, 0.05% of EDTA disodium, 0.3% of hyaluronic acid, 20% of cosmetic raw materials, 0.01% of citric acid, 0.03% of sodium citrate and the balance of water; preparing a water phase pot: heating and uniformly mixing deionized water and cosmetic raw materials according to the formula amount, and stirring to obtain a water phase pot;
(II) weighing butanediol and xanthan gum according to the formula, stirring uniformly in advance, and adding into a water phase pot;
(III) weighing the glycerol, EDTA disodium and hyaluronic acid according to the formula amount, adding the mixture into a water phase pot, stirring uniformly to dissolve powder substances in the liquid, preserving heat, and cooling in a cold water bath;
and (IV) adding the citric acid and the sodium citrate in the formula amount, stirring, dissolving, uniformly mixing, and homogenizing to obtain the distilled grain cosmetic water of the white spirit.
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| CN117547498A (en) * | 2024-01-11 | 2024-02-13 | 珠海远大美业生物科技有限公司 | Distilled grain skin beautifying liquid and preparation method and application thereof |
| CN117547498B (en) * | 2024-01-11 | 2024-04-26 | 贵州科益创生物科技有限公司 | Distilled grain skin beautifying liquid and preparation method and application thereof |
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| CN117547498A (en) * | 2024-01-11 | 2024-02-13 | 珠海远大美业生物科技有限公司 | Distilled grain skin beautifying liquid and preparation method and application thereof |
| CN117547498B (en) * | 2024-01-11 | 2024-04-26 | 贵州科益创生物科技有限公司 | Distilled grain skin beautifying liquid and preparation method and application thereof |
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