CN108143678A - Sorghum vinasse extract - Google Patents
Sorghum vinasse extract Download PDFInfo
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- CN108143678A CN108143678A CN201711280479.9A CN201711280479A CN108143678A CN 108143678 A CN108143678 A CN 108143678A CN 201711280479 A CN201711280479 A CN 201711280479A CN 108143678 A CN108143678 A CN 108143678A
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- sorghum
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- vinasse
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- 239000000284 extract Substances 0.000 title claims abstract description 58
- 235000011684 Sorghum saccharatum Nutrition 0.000 title claims abstract description 48
- 241000209072 Sorghum Species 0.000 title description 3
- 240000006394 Sorghum bicolor Species 0.000 claims abstract description 45
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 33
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000000203 mixture Substances 0.000 claims abstract description 15
- 239000002537 cosmetic Substances 0.000 claims abstract description 13
- 239000002904 solvent Substances 0.000 claims abstract description 10
- 239000012530 fluid Substances 0.000 claims abstract description 8
- 239000000654 additive Substances 0.000 claims abstract description 7
- 230000000996 additive effect Effects 0.000 claims abstract description 7
- 230000003078 antioxidant effect Effects 0.000 claims abstract description 5
- 230000002087 whitening effect Effects 0.000 claims abstract description 5
- 239000002994 raw material Substances 0.000 claims abstract description 3
- 230000003013 cytotoxicity Effects 0.000 claims abstract 3
- 231100000135 cytotoxicity Toxicity 0.000 claims abstract 3
- 238000000605 extraction Methods 0.000 claims description 32
- 235000019441 ethanol Nutrition 0.000 claims description 19
- 230000003647 oxidation Effects 0.000 claims description 8
- 238000007254 oxidation reaction Methods 0.000 claims description 8
- 239000003814 drug Substances 0.000 claims description 7
- 230000036564 melanin content Effects 0.000 claims description 7
- 238000000194 supercritical-fluid extraction Methods 0.000 claims description 7
- 238000000638 solvent extraction Methods 0.000 claims description 6
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
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- 150000001298 alcohols Chemical class 0.000 claims 1
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- XUMBMVFBXHLACL-UHFFFAOYSA-N Melanin Chemical compound O=C1C(=O)C(C2=CNC3=C(C(C(=O)C4=C32)=O)C)=C2C4=CNC2=C1C XUMBMVFBXHLACL-UHFFFAOYSA-N 0.000 abstract description 12
- -1 has low cytotoxicity Substances 0.000 abstract description 11
- 102000003425 Tyrosinase Human genes 0.000 abstract description 8
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- 238000004458 analytical method Methods 0.000 description 20
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- 230000031700 light absorption Effects 0.000 description 12
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- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 4
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- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
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- DWNBOPVKNPVNQG-LURJTMIESA-N (2s)-4-hydroxy-2-(propylamino)butanoic acid Chemical compound CCCN[C@H](C(O)=O)CCO DWNBOPVKNPVNQG-LURJTMIESA-N 0.000 description 1
- KSEBMYQBYZTDHS-HWKANZROSA-M (E)-Ferulic acid Natural products COC1=CC(\C=C\C([O-])=O)=CC=C1O KSEBMYQBYZTDHS-HWKANZROSA-M 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
-
- 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
- A61K2800/74—Biological properties of particular ingredients
- A61K2800/78—Enzyme modulators, e.g. Enzyme agonists
- A61K2800/782—Enzyme inhibitors; Enzyme antagonists
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Dermatology (AREA)
- Microbiology (AREA)
- Epidemiology (AREA)
- Birds (AREA)
- Mycology (AREA)
- Botany (AREA)
- Biotechnology (AREA)
- Engineering & Computer Science (AREA)
- Gerontology & Geriatric Medicine (AREA)
- Cosmetics (AREA)
- Medicines Containing Plant Substances (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention discloses a sorghum vinasse extract, which is obtained by extracting sorghum vinasse by using Jinmen sorghum vinasse as a raw material and using pure water, an alcohol solvent and supercritical fluid; the sorghum vinasse extract has antioxidant activity, tyrosinase inhibiting ability and melanin generation reducing ability, and can be used as additive for preparing makeup or medicinal makeup composition; wherein the sorghum vinasse extract is present in an amount of 0.001 to 0.003 wt% based on the total weight of the make-up or cosmeceutical composition, has low cytotoxicity, antioxidant capacity and whitening capacity.
Description
Technical field
The present invention relates to a kind of sorghum vinasse extracts, can be used as especially with regard to one kind and prepare makeups or medicine composition for cosmetics
Additive purposes sorghum vinasse extract.
Background technology
Sorghum vinasse (sorghum distillery residue, SDR) are generated by-product after brewing liquor made from sorghum,
8.2 kilograms of sorghum vinasse can be about generated after 25 kilograms of sorghum-brewed wines, and Jinmen brewery sorghum vinasse output per day is up to 250 public affairs
Ton is mainly provided as Jinmen locality and cooks feed stripped or be to provide general family or small scale-farm as organic at present
Fertilizer.But aforementioned carried usage amount is very little always, the sorghum vinasse that can be digested are fairly limited, therefore most liquor made from sorghum
Grain is all still not utilized and is handled with trade waste, and the preparation method of general liquor made from sorghum is steamed three for three and made, and life matter utilizes
Rate is about 9 one-tenth, and the liquor made from sorghum of Jinmen brewery is prepared as two steamings two and makes, and life matter utilization rate is less than 7 one-tenth, and it is desirable to by this substance
Its functional ingredient is extracted using green non-poisonous prepare, applied on makeups skin care products composition.
According to the by-product vinasse that Jinmen brewery research sorghum in 2001 was fermented, it is (dry that content of starch still may be up to 25%
Weight) more than, and document is pointed out, polyphenol compound (polyphenol of the sorghum with abundant life, pharmacological activity
Compounds), such as tannin (tannins), flavone compound (flavonoids), phenolic acid (phenolic acid), and
Other functional ingredients also have long-chain fatty alcohol (policosanols) and phytosterol (Phytosterol) etc..Research confirms
There is these functional ingredients removing interior free yl, lipid peroxidation inhibition, prevention of cardiovascular disease, reduction human serum courage to consolidate
Alcohol, anticancer and it is antimicrobial the effects that.Also have studies have shown that snakeheaded fish ingest addition sorghum vinasse feed can enhance blood moderate resistance
Oxidation characteristic, and the solidification of blood can be reduced, with the characteristic of cold-resistant, increase the rate of growth in winter.
It is as follows for the research of sorghum vinasse in the document of previous publications.TaiWan, China patent I430755, a kind of tool
There are the sorghum vinasse extract and its manufacturing method of antibacterial：Following extraction mode, the extraction of (a) cold water are used in this patent：
At 0~25 DEG C, sorghum vinasse and distilled water are stirred with the proportional region of 1/10~1/100w/v；(b) alcoholic extract：0
At~25 DEG C, sorghum vinasse and 5~95% alcoholic solutions are stirred with 1/10~1/100w/v proportional regions；(c) water hot extraction：
Sorghum vinasse and distilled water are obtained with 1/10~1/100w/v proportional regions through 100~135 DEG C of heat treatments selected from the above method
Extract after, then be filtered, after removing moisture removal, be milled up to sorghum vinasse powder, because its powder is without directly mill
Powder and without the fibrous matters such as paddy and sorghum husk, its object is to do antibacterial purposes, inhibit Bacillus cereus (Bacillus
Cereus), Escherichia coli O 157:H7(Escherichia coli O157:H7), salmonella (Salmonella spp.) and
The effect of bacterial strains such as aurococcus (Staphylococcus aureus), therefore can make as food additives
With.
Taiwan agriculture chemistry and Food Science (the TaiWan, China fifth phase of volume 48 in October, 99：2nd 33
To page 23 8), the extraction of sorghum vinasse functional ingredient, this document purpose, which is to extract, to be prepared using different solvents for extraction
Take the influence of ingredient, utilize (1) methanol, isopropanol, n-hexane, acetone, ethyl acetate, different proportion ethanol water and
90 DEG C of hot water equal solvents are stirred extraction；(2) supercritical carbon dioxide extraction sorghum vinasse, extraction temperature are 35~80
DEG C and extracting pressure be 172~483 bars, carry out 3 hours extract.
TaiWan, China patent publication No. 201000628 is disclosed and is prepared with anti-oxidant and inhibition tyrosinase activity
The method of health material, main purpose is that its exploitation is prepared by Co 2 supercritical fluid separable programming extraction in this patent
Take the yellow rice wine dregs of rice, operation temperature is 40 DEG C, 350 bars of pressure when, and the weight ratio of carbon dioxide usage amount amount of solvent together
Less than 40, can obtain with anti-oxidant and inhibition tyrosinase activity health material.
TaiWan, China patent the 104142021st, discloses vinasse extract and its manufacturing method, utilizes acid, basic hydrolysis
It to destroy plant cell wall method, then detaches via ethanol purification, in addition to it can extract and obtain free property phenolic acid, and combination can be increased
The acquisition content of property phenolic acid, to increase yield.Wherein vinasse extract is via high-performance liquid chromatograph (High-Performance
Liquid Chromatography, HPLC) identify its ingredient after, then via gas chromatography-mass spectrography
(chromatography-mass spectrometry, GC-MS) carries out Structural Identification, it was demonstrated that it includes a variety of phenol such as ferulic acid
Sour functional ingredient, such compound have oxidation resistance, the white-skinned face function for inhibiting tyrosinase, can be applied to cosmetology
Product composition.
The present invention is further using the extraction the preparation method of complete green non-poisonous, harmless chemical solvent, and disclosed in the invention system
Under standby, most adapt to be used to prepare the most suitable adding proportion of the additive of makeups or medicine composition for cosmetics.
Invention content
The present invention using solvent extraction in addition to (using solvent as under the ethanol water and different temperatures of different proportion
Pure water) it carries out outside the extraction of sorghum vinasse, also extracted using supercritical carbon dioxide extraction equipment.The system of the present invention
Preparation Method is the functional ingredient extraction applied to skin care products, therefore considers can be applied to the group component requirements of skin care products, is being used
During solvent extraction, it is necessary to it avoids using the organic solvent that can be damaged to environment and human body, such as：Methanol, n-hexane etc.,
It is ethyl alcohol and pure water to be selected, and is extracted, and with the United States Pharmacopeia USP of high gauge by its ratio of regulation and control and temperature
The specification of 467 dissolvent residuals carries out the self-examination in raw material exploitation, and ethyl alcohol is III grade of residual solvent of its specification, generally recognize
It compares low toxicity, low-risk for I grade and II grade of residual solvent for III grade of residual solvent harm to the human body.
No doubt it is common method in addition to using solvent extraction, is in addition also set using supercritical carbon dioxide extraction
It is standby, using carbon dioxide as the fluid of supercritical extraction technique, meet the standard of green industry preparation, be also apt to " corporate social to the greatest extent
Responsibility CSR (Corporate Social Responsibility) ", and meet in various countries' health regulation, makeup maintenance product
The regulation of harmful to human substance must not be remained, in addition extraction temperature is low, can preferably retain the compositions such as the bioactivity of extract.
Along with using co_2 supercritical fluid extraction, supercritical fluid has selectivity, and the experimental design by the present invention is tested
Card can obtain the vinasse functional Ingredients Active object that purity is high, toxicity is low, be applicable to the application as cosmetics.
The sorghum that the present invention greatly utilizes kaoliang spirit steams the waste generated after wine, it is expected to change in sorghum vinasse waste
Into available substance, economic value can more be promoted by not only reducing environmental pollution.And environmental consciousness comes back in recent years, and evil mind
Commodity problem getting worse, the safety for being in direct contact the skin care products and color cosmetic of human skin can not be ignored, in view of general
Commercially available skin care products and color cosmetic are mostly containing preservative and artificial essence and pigment etc., if extracted in the preparation of Material synthesis pure
That changes is incomplete, causes byproduct residue in the feed, may result in after long-time service human body accumulate many chemical analysis and
The skin damages such as sensitive, stimulation are caused, therefore consumer more advocates natural keep-fit product, therefore the present invention is wished to develop and can be allowed
Consumer feel at ease the makeup used maintenance product.
Present invention research and development are confirmed compared with functional ingredient in the preparation extraction sorghum vinasse of natural environmental-protective via experiment
The Determination of Polyphenols that its solvent extraction obtains extract is about 207.46mg/100g SDR to 298.87mg/100g SDR,
And the Determination of Polyphenols that supercritical carbon dioxide extraction method obtains extract is about 27.22mg/100g SDR, is previously ground
Studying carefully confirms these polyphenolic substances, has oxidation resistance and has inhibiting effect to Tyrosine Enzyme, and is not useful for makeup maintenance
The precedent of product, therefore the concentration for being best suited for cosmetics is found out using this extract liquor obtains it in cosmetic composition most
Be suitble to content for 0.001 weight % to 0.02 weight %, and its in cosmetic composition content be 0.02 weight % when, tool
There is more than 80% oxidation resistance.
Description of the drawings
Fig. 1 is that 1,1- hexichol -2- picryls hydrazines (DPPH, 1,1-Diphenyl-2-picrylhydrazyl) remove free radical
The analysis result that ability measures；
Fig. 2 is the analysis result of MTT cell survival rates；
Fig. 3 is the analysis result of intracellular tyrosine enzymatic activity；
Fig. 4 is the analysis result for generating melanin content reduction.
Specific embodiment
Below by the preparation of specific embodiment discussion sorghum vinasse extract liquor and its purposes applied to skin-care product.
Persond having ordinary knowledge in the technical field of the present invention, can be understood as the content disclosed by this specification the present invention other are excellent
Point and effect, and the embodiment discussed under without departing substantially from scope of the invention the present invention is modified and is changed, and still belong to
In the scope of the present invention.
It is prepared by 1 sorghum vinasse extract liquor of embodiment
(1) sorghum vinasse：Test specimen recycles powder through using fluidized bed continuous drying equipment at 100 DEG C to 120 DEG C
Broken machine is ground.
(2) solvent extraction sorghum vinasse：20g sorghums vinasse are weighed in triangular pyramidal flask, respectively with 75 DEG C of 200ml
Pure water, 90 DEG C of pure water, 75% ethyl alcohol and 90% ethanol water carry out the stirring extraction of 1 hour, and extraction ends again with filter paper mistake
Quantitative filtrate volume after filter.
(3) supercritical carbon dioxide extraction sorghum vinasse：Supercritical carbon dioxide extracting used in the present invention makes
It is carried out with the extraction tank of 250ml.It extracts three times and takes sorghum vinasse powder 93.51g, 98.85g and 98.85g respectively, open dioxy
Change carbon to enter in pipeline, adjustment counterbalance valve (back pressure regulaor) control system pressure (extracting pressure point three times
Wei not be 170 bars, 250 bars and 300 bars) and adjustment extraction tank in temperature (this time temperature in use be 40 DEG C), the co-extraction used
Solvent is ethyl alcohol, starts timing after pressure and temperature reach experiment setting value, first surpasses sample using ethyl alcohol and carbon dioxide
Critical fluids are soaked in extraction tank 2 hours with static extracting, later with the carbon dioxide of flow velocity 0.2L/hr and flow velocity 0.4L/hr
Ethyl alcohol carry out dynamic extraction 2 hours, the sample collected by two separating tanks is uniformly mixed, is saved backup.
2 total polyphenols compound of embodiment (Total phenolic compounds, TPC) content analysis
(1) standard items and preparation of reagents：Standard items are prepared：This experiment can utilize prior art method, with gallic acid (gallic
Acid, Arcos Organics) for standard items, the anhydrous gallic acid for weighing 0.1g adds in 100ml pure water with ultrasonic oscillation
After being uniformly dissolved, make standard concentration for 1000ppm, then the volume indicated according to following table, add in pure water serial dilution various concentration
Standard items：
| Concentration (mg/L)；ppm | 0 | 10 | 20 | 30 | 40 | 50 |
| 1000ppm standard items (mL) | 0 | 1 | 2 | 3 | 4 | 5 |
| Pure water (mL) | 100 | 99 | 98 | 97 | 96 | 95 |
Experiment reagent then needs preparation (a) Folin reagent (Folin-ciocalteus phenol reagent) dilute with pure water
Release 10 times spare (b) 7.5% aqueous sodium carbonate (Na2CO3)。
(2) experimental procedure：Standard items and sample are configured to debita spissitudo with pure water, then 1ml is taken to add in 5ml dilutions 10 respectively
Folin reagent again, is protected from light standing 5 minutes after being mixed evenly, add in 7.5% aqueous sodium carbonate of 4ml later, be mixed evenly
After be protected from light standing 1 hour after centrifuge, its light absorption value is measured under 765nm with spectrophotometer.Using standard concentration as abscissa,
Light absorption value does linear regression analysis for ordinate, when calibration curve coefficient correlation (R2) can be used for 0.999 to 1 side.Later again
Sample light absorption value substitution standard curve is calculated into its total polyphenols compounds content.The extraction quantity of total polyphenols compound is with every hectogram height
Beam vinasse extract the milligram number and represent (mg/100g SDR).
(3) experimental result：
Table one is the analysis result of Determination of Polyphenols, and when pure water is used to extract, temperature has no significant effect it, but
It is when being extracted using 75% ethyl alcohol, Determination of Polyphenols is apparently higher than 90% alcohol extraction liquid, and uses supercritical extract mode,
Determination of Polyphenols then extracts mode significantly lower than other two kinds, and because using carbon dioxide, for solvent, polarity is relatively low, and Polyphenols
For compound because containing hydroxyl, the carbon dioxide compatibility that most of polarity is all higher and polarity is relatively low is poor.
Table one
| Determination of Polyphenols (mg/100g SDR) | |
| 90 DEG C of pure water extractions | 227.3±3.7 |
| 75 DEG C of pure water extractions | 207.5±1.2 |
| 90% ethyl alcohol extracts | 209.7±1.7 |
| 75% ethyl alcohol extracts | 298.9±12.8 |
| The sample that supercritical extract merges three times | 27.2±0.39 |
Embodiment 3 is removed DPPH (1,1-Diphenyl-2-picrylhydrazyl) free radical ability and is measured
(1) preparation of reagents：The DPPH of 0.004g is taken to add in the ethyl alcohol (a concentration of 40ppm) of 100ml, is protected from light after uniform dissolution
It saves backup, and stores and may not exceed half a day.
(2) experimental procedure：Each extract liquor is configured to 200ppm, then with solvent of its extraction be sequentially diluted to 100ppm,
50ppm, 25ppm and 12.5ppm take the DPPH of 6ml to add in 1ml samples, standing are protected from light after being mixed evenly 40 minutes, to be divided light
Degree meter measures its light absorption value, and the clean-up effect percentage of the DPPH free radicals according to each extract liquor of following equation calculating under 517nm
Than (scavenging effects；%)
The clean-up effect percentage (%) of DPPH free radicals
(3) experimental result：
Contained substance is complex in extract liquor, and phenolic compound has high correlation with oxidation resistance, therefore removes
DPPH free radical ability determination experiments are using total polyphenols concentration as foundation.Fig. 1 is scavenging ability of DPPH free radical determination experiment
Analysis result, abscissa are then the content of total polyphenols compound；Fig. 1 shows its Scavenging activity as extract liquor concentration declines and drops
It is low and best with 75% alcohol extraction liquid and 90% alcohol extraction liquid, have under the concentration of 200ppm about 85.3% and
85.2% Scavenging activity.
Due to mainly having the substance of free radical scavenging ability in sorghum vinasse for total polyphenols compound, also demonstrate,proved by experimental result
Positive correlation is presented with total polyphenols compound in the DPPH free radical scavenging abilities of bright extract liquor.Thus experimental result understands vinasse extraction
Liquid sample can act on anti-aging, oxidation resistant application..
The analysis of 4 cell survival rate of embodiment
(1) experimental principle
It can also be originated in by the dehydrogenase in grain wire body in living cells using tetrazole (Tetrazolium) in MTT reagents
The characteristic that raw bluish violet formazan (Formazan) crystallizes, to detect the relative scale of survivaling cell.Formazan crystal is in wavelength
There is maximum light absorption value during 570nm, therefore the survival rate of cell is calculated by the measure of light absorption value.
(2) experimental method
By melanoma cells (5 × 104Cells/well) it is implanted into 24 hole culture plates, after cultivating 24 hours, with final concentration
The vinasse extraction solution example processing cell of 10ppm, 20ppm, 30ppm 24 hours, it is primary with PBS cleanings per hole, add in MTT
After solution is placed in 37 DEG C, 4 hours, is dissolved using dimethyl sulfoxide (DMSO) (DMSO) and MTT is restored by the dehydrogenase in cell Mitochondria
Formation formazan products measure the OD570 light absorption values of product with enzyme-linked immunosorbent assay (ELISA).Cell survival is the more then
Coloured product is more dark purple.
Cell survival rate formula (%)=100- [(A-B)/A × 100%]
A：The OD570 light absorption values of control group
B：The OD570 light absorption values of sample sets
(3) experimental result：
Fig. 2 is the cell survival rate analysis of water extraction, alcoholic extract and supercritical extract, and final concentration in the solution is distinguished
For 10ppm, 20ppm, 30ppm.By experimental result it is found that supercritical fluid has preferable cell in identical vinasse extraction solution
Survival rate, and vinasse extract liquor, in addition to alcohol 30ppm, remaining sample cell survival rate is all more than 70%.Thus experimental result
Understand vinasse extract liquor sample will not to cell toxic effect.
The analysis of 5 intracellular tyrosine enzymatic activity of embodiment
The analysis principle of intracellular tyrosine enzymatic activity
Using the method for Frozen-thawed cycled (Freeze-thaw cycle), cell is placed in -80 DEG C of freezing a period of times (30
Minute), intracellular moisture is made to form ice crystal；Cell taking-up is placed in thaw at RT (30 minutes) again, ice crystal is made to revert to water
Shape makes cell rupture, and then release intracellular organic matter by this process, and the tyrosinase that we to be measured is also in wherein.Add
Enter the catalytic receptor (such as levodopa (L-dopa)) of tyrosinase, analyze the production quantity of its product, can deduce its tyrosinase
Relative activity.
The analysis method of intracellular tyrosine enzymatic activity
By dopachrome (dopachrome) autoxidation inhibiting rate is measured, to understand whether vinasse extract liquor can inhibit junket
Propylhomoserin enzymatic activity and reduce melanin and formed.The implant density 5 × 10 in 24 hole culture plates4The B16F10 melanin of cells/well
Oncocyte is incubated in the culture medium of alpha-melanocyte stimulating hormone containing 100nM (α-MSH), is placed in 37 DEG C, 5%CO2In incubator
After being kept for 24 hours, add in vinasse supercritical fluid extraction (SFE) and continue with 24 hours, 100 μ l 1% are then added in per hole
(V/V) Triton X-100 (Triton X-100)/50mM sodium phosphate buffers (pH 6.8) and 10mM benzyl sulphurs
Acyl fluorides (PMSF) is placed in -80 DEG C, 30 minutes, further takes out in room temperature 30 minutes, after being centrifuged 30 minutes with 12000r.p.m rotating speeds, takes
Go out 80 μ l upper stratas suspension to be placed in 96 porose discs, and add in the 1mg/ml DOPA solution of 20 μ L per hole, measure OD490 extinctions
Value measures once, a continuous hour, finally calculates tyrosinase inhibition rate again for every 10 minutes.
Inhibiting rate formula (%)=(A-B)/A × 100%
A：The OD490 light absorption values of control group
B：The OD490 light absorption values of sample sets
Experimental result
Fig. 3 is the analysis result of intracellular tyrosine enzymatic activity, and the vinasse extract liquor of supercritical extract has preferable junket ammonia
Sour enzyme inhibition rate, in the solution when a concentration of 20ppm, 30ppm, inhibiting rate is up to more than 70%.Thus experimental result understands wine
Poor extract liquor sample can act on the application of whitening efficiency.
The analysis of 6 melanin content of embodiment
The analysis principle of melanin content
Using high temperature and highly basic intracellular melanin is dissolved out, and divide in the measure of its special extinction wavelength 405nm
Analyse influence of the processing of various different conditions to melanin content in B16F10 melanoma cells.
(2) analysis method of melanin content
5 × 10 are implanted into per hole in 24 hole culture plates4A cell is incubated in the culture medium containing 100nM α-MSH, culture
24 hours (37 DEG C, 5%CO2In incubator).Vinasse SFE or kojic acid of the rear substitution containing various concentration culture medium (control group
Only change culture medium), it is further cultured for 24 hours.Then culture medium is removed, 100 μ l 1N NaOH (10% are added in after being cleaned 2 times with PBS
DMSO is prepared) it is reacted 1 hour in 60 DEG C, it finally takes and removes the light absorption value for measuring wavelength 405nm, meter in 80 μ l to 96 porose discs in every hole
Calculate the relative amount of melanin.
(3) experimental result
Fig. 4 is the analysis result of melanin content, and the vinasse extract liquor sample of water extraction, alcoholic extract and supercritical extract is all
The efficiency of generation melanin is reduced, and wherein has preferable result with supercritical extract 30ppm.Thus experimental result is understood
Vinasse extract liquor sample can act on the application of light scar, light spot.
Claims (10)
1. a kind of sorghum vinasse extract, by with sorghum vinasse, particularly Jinmen sorghum vinasse for raw material, with pure water or alcohol
Class solvent or supercritical fluid extraction sorghum vinasse obtain.
2. the extract as described in claims the 1st, wherein, extraction temperature is 50 DEG C to 99 DEG C, and extraction time is small for 2
When.
3. the extract as described in claims the 1st, wherein, which is ethyl alcohol, the ethyl alcohol a concentration of 50% to
100%, extraction time is 2 hours, 30 DEG C of extraction temperature.
4. the extract as described in claims the 1st, wherein, which includes：First by sorghum vinasse original
It is continuous to reuse Co 2 supercritical fluid using ethyl alcohol and Co 2 supercritical fluid static extracting 2 hours later for material
Dynamic extraction 2 hours, the dried moisture content of sorghum vinasse used are less than 30 weight %, and operating pressure is between 150 bars to 350
Between bar, temperature is between 40 DEG C to 60 DEG C.
5. the extract as described in claims the 4th, the supercritical fluid extraction optimum condition include：Operating pressure 170
Bar, 40 DEG C of temperature；170 bars of operating pressure, temperature 50 C；170 bars of operating pressure, 65 DEG C of temperature；250 bars of operating pressure, temperature
40℃；And 300 bars of operating pressure, 40 DEG C of temperature.
6. the extract as described in any one of claims 1-5, which has antioxidant activity, inhibits junket ammonia
Sour Enzyme abilities and reduction generation melanin content ability, can be as the additive purposes for preparing makeups or medicine composition for cosmetics.
7. the extract as described in claims the 6th, the extract is as the additive for preparing makeups or medicine composition for cosmetics
Purposes, to prevent skin aging, whitening, tender white, light spot or bright skin.
8. the extract as described in claims the 7th, the extract as the additive for preparing makeups or medicine composition for cosmetics,
The extract exists with the amount of 0.001 weight % of the total weight based on the composition to 0.02 weight %.
9. the extract as described in claims the 7th or 8, the extract is as preparing makeups or the addition of medicine composition for cosmetics
Object, wherein, which exists with the amount of 0.001 weight % of the total weight based on the composition to 0.003 weight %, have compared with
Low cytotoxicity, oxidation resistance and whitening capability.
10. the extract as described in any one of claims 7-9, the extract is as preparing makeups or the combination of medicine adornment
The additive of object, wherein having minimum cytotoxicity, oxidation resistance and whitening energy with the extract of supercritical fluid extraction
Power.
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| TW106125807A TWI653056B (en) | 2016-12-06 | 2017-07-28 | Use of sorghum distiller's extract as whitening skin care product |
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| CN110305750A (en) * | 2019-07-03 | 2019-10-08 | 富民县梓源贸易有限责任公司 | A kind of red sorghum wine and its brewing method |
| CN110960474A (en) * | 2019-12-27 | 2020-04-07 | 爱思开百朗德生物科技（海门）有限公司 | Anti-inflammatory, antioxidant and anti-wrinkle rice grain essence and preparation method thereof |
| CN112998181A (en) * | 2021-03-18 | 2021-06-22 | 中国科学院近代物理研究所 | High-activity sweet sorghum polyphenol and preparation method and application thereof |
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| TW201821059A (en) | 2018-06-16 |
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