CN106038411B - Multi-form melanin precursor substance hair dye and its application method - Google Patents
Multi-form melanin precursor substance hair dye and its application method Download PDFInfo
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
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- 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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/40—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing nitrogen
- A61K8/44—Aminocarboxylic acids or derivatives thereof, e.g. aminocarboxylic acids containing sulfur; Salts; Esters or N-acylated derivatives thereof
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- 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
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/49—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds
- A61K8/4906—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom
- A61K8/4913—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having five membered rings, e.g. pyrrolidone carboxylic acid
- A61K8/492—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds containing heterocyclic compounds with one nitrogen as the only hetero atom having five membered rings, e.g. pyrrolidone carboxylic acid having condensed rings, e.g. indol
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q5/00—Preparations for care of the hair
- A61Q5/10—Preparations for permanently dyeing the hair
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- 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
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Abstract
The invention discloses a kind of two dosage form melanin precursor substance hair dyes and a kind of preparation method of three dosage form melanin precursor substance hair dyes.Two component hair dyes include: A agent and B agent；Three component hair dyes include: A agent, B agent and C agent；Wherein A agent and C agent include melanin precursor, hair dye usual auxiliaries and water；Hair dye usual auxiliaries are one of wetting and penetrating agent, thickener, surfactant, moisturizer, pH adjusting agent or a variety of.In conjunction with the advantages of synthetic hair dyes and natural hair dye, by extracting the complex enzyme liquid in natural edible bacterium, amino acid is synthesized into dopachrome, 5,6- dihydroxy indole and 5 by bioconversion, the melanin precursors substances such as 6- dihydroxy indole -2- carboxylic acid are prepared into biology hair dyeing agent method.Have many advantages, such as during production and use raw material be easy to get, simple process, using effect it is reliable and safe and non-toxic and environmentally friendly, be easy to the factorial production, there is huge application value in hair dye preparation field.
Description
Technical field
The present invention relates to a kind of utilizations to extract the preparation method that biological complex enzyme catalyzes and synthesizes melanin precursor object hair dye,
More particularly to a kind of extraction edible mushroom complex enzyme liquid, amino acid substrate is converted to natural black pigment precursor substance, and by its
It is prepared into Non-oxidized multi-form biology hair dye.
Background technique
Currently, the continuous improvement with people to healthy attention degree, contain in chemically synthesized hair dye carcinogenic and
Strong sensitization chemistry staining components (such as phenylenediamine, amino phenols substance) and some harmful components (such as hydrogen peroxide, ammonium hydroxide),
It is concerned by people more and more.For this purpose, being attempted to seek natural harmless, safer dye material.Such as
Phytochrome hair dye, although achieving certain achievement, but still there are many problems, this kind of product, which not can avoid still, to be contaminated
It sends out and adds the harmful substances such as phenols and hydrogen peroxide in agent, and lack mature pigment refinement technique.It has been currently known utilization
Phytochrome catechol prepares the method (patent document CN101164528A) of natural hair dye, but catechol used in it is
It is extracted from Chinese medicine catechu, the disadvantage is that the planting scale of Chinese medicine is limited, pigment extraction cost is relatively high；Also it has been reported that
By technique for gene engineering, using the catechol-oxydase in expression mould category der Pilz cell, to DOPA and its derivative into
Row oxidation, is transformed into the method (patent document CN101914585A) of melanin precursor, but mould itself can generate
Poisonous and harmful substance, production process will cause the pollution of environment, exist simultaneously at high cost, and time-consuming, and performance is unstable, Wu Fa great
The disadvantages of amount application.Also have before generating melanin using enzyme solution catalysis substrate extracted in fermentation of bacillus liquid in the prior art
The method of body substance, but still not can guarantee the safe without toxic side effect of its product, the pollution in fermentation process also will affect product
Quality.
The color of human hair is determined by the type and quantity of the melanin granule in cortex of hair, with the age
Growth and allegro Working Life caused by pressure, the generation of melanin granule can gradually decrease, and dark hair will gradually become
It is white.And the synthesis process (Fig. 1) of melanin in human body, it is to pass through tyrosinase by melanocyte for oxidizing tyrosine at more
Bar pigment, dopachrome decarboxylize generates 5,6- dihydroxy indole, or generates 5,6- dihydroxy indole-using tautomerase
2- carboxylic acid forms eumelanin by polymerization later, to make hair that black be presented.
Summary of the invention
The present invention is in order to solve the above-mentioned technical problem, and according to the composition principle of melanin in human hair, to provide
One kind extracting enzyme solution inside edible mushroom, generates melanin precursor substance by reacting with amino acid substrate, auxiliary material is then added
Multi-form biology hair dye is prepared, process includes: extraction, purifying, fixation and the preservation work of (a) edible mushroom complex enzyme liquid
Sequence；(b) substrates such as tyrosine are subjected to biotransformation into the process of melanin precursor；(c) above-mentioned reaction solution is handled, is mentioned
The process of high 5,6- dihydroxy indole concentration；(d) it is mixed with melanin precursor substance and auxiliary material to obtain multi-component hair dye
Process.
Two dosage form melanin precursor substance hair dyes of the invention, the hair dye includes: A agent and B agent；
The A agent includes melanin precursor, auxiliary agent A and water；The auxiliary agent A is wetting and penetrating agent, thickener, surface-active
One or more of agent, moisturizer, pH adjusting agent；
The melanin precursor includes dopachrome, 5,6- dihydroxy indole, one in 5,6- dihydroxy indole -2- carboxylic acid
Kind is several；
The weight percent that the melanin precursor accounts for A agent is 0.01%~5%；
The B agent includes auxiliary agent B and water；The auxiliary agent B is dyeing auxiliaries, in thickener, surfactant, pH adjusting agent
One or more.
Preferably, the wetting and penetrating agent is one or both of L-cysteine, glutathione；
The dyeing auxiliaries is the metallic salt containing divalent or Tricationic；
The thickener is sodium carboxymethylcellulose, carboxymethyl cellulose, carboxyethyl cellulose, starch, Silica hydrogel, bright
Glue, sodium alginate, guar gum, agar, soybean protein glue, polyethylene glycol, Sodium Polyacrylate, polyacrylamide, poly- γ-paddy ammonia
One or more of acid, hyaluronic acid sodium, polyacrylic acid, low-molecular polyethylene wax, xanthan gum；
The surfactant is glycerin monostearate, hexadecyltrimethylammonium chloride, cetanol, lauryl sulfonation
One or more of monomester succinate disodium, sulfosuccinic acid monoesters disodium, love Ma cream；
The moisturizer is one or more of glycerol, sorbierite, mannitol, ethylene glycol, glycerine, edible oil；
The pH adjusting agent is one or more of lysine, arginine, sodium bicarbonate.
Preferably, the divalent or Tricationic are Cu2+、Mg2+、Ca2+、Fe2+、Fe3+One or more of.
Preferably, in the A agent:
The weight percent that the wetting and penetrating agent accounts for A agent is 1%~10%；
The weight percent that the thickener accounts for A agent is 0.2%~2%；
The weight percent that the surfactant accounts for A agent is 0.1%~2%；
The weight percent that the moisturizer accounts for A agent is 1%~15%；
The weight percent that the pH adjusting agent accounts for hair dye is 1%~10%；
In the B agent:
The weight percent that the dyeing auxiliaries accounts for B agent is 0.5%~10%；
The weight percent that the thickener accounts for B agent is 0.2%~2%；
The weight percent that the surfactant accounts for B agent is 0.1%~2%；
The weight percent that the pH adjusting agent accounts for B agent is 0.1%~1%.
Preferably, the melanin precursor preparation the following steps are included:
(1) prepare reaction substrate；
(2) complex enzyme needed for extracting synthesis of melanin precursor in edible mushroom；
(3) reaction substrate in step 1 is catalyzed using complex enzyme made from step 2, obtains target product；
(4) gained target product in step 3 is subjected to conversion processing, obtains melanin precursor；
The reaction substrate is one of tyrosine, DOPA, dopamine or a variety of；
The edible mushroom be one of Pleurotus eryngii, agaricus bisporus, brown mushroom, straw mushroom, oyster mushroom, mushroom, black fungus, agrocybe or
It is a variety of.
Preferably, the target product conversion processing one of performs the following operation or a variety of: boiling water heating, heating are same
When be evaporated under reduced pressure, electrolysis, addition decarboxylase.
Preferably, the substrate is l-tyrosine.
Preferably, the concentration of substrate is 1mM~10mM.
Preferably, the complex enzyme extracted in the step 2 carries out purifying and immobilization processing；The method of the purifying is
Ultrafiltration and/or ion-exchange chromatography.
Preferably, described when immobilizing processing to complex enzyme, fixed material is molecular sieve, diatomite, sodium alginate
Deng well known immobilization material.
The application method of two dosage form melanin precursor substance hair dyes of the invention, the application method the following steps are included:
The A agent of above-mentioned two dosage forms melanin precursor substance hair dye will be uniformly applied to hair surface after dyeing hair cleaning, 30~
It is cleaned after 50min with clear water；B agent is uniformly applied to hair surface again, after 15~25min, is cleaned, i.e., dyed hair with clear water
Black.
Preferably, the usage ratio of the A agent and B agent is 3:1~1:1, and the ratio is mass ratio.
Three dosage form melanin precursor substance hair dyes of the invention, the hair dye includes: A agent, B agent and C agent；
The A agent includes melanin precursor, auxiliary agent A and water；Melanin precursor accounts for the weight percent of A agent in the A agent
It is 0.01%~5%；The auxiliary agent A is wetting and penetrating agent, thickener, surfactant, moisturizer, one in pH adjusting agent
Kind is several；
The B agent includes auxiliary agent B and water；The auxiliary agent B is dyeing auxiliaries, in thickener, surfactant, pH adjusting agent
One or more.
The C agent includes melanin precursor, auxiliary agent C and water；Melanin precursor accounts for the weight percent of C agent in the C agent
It is 0.01%~5%；The auxiliary agent C is wetting and penetrating agent, thickener, surfactant, moisturizer, one in pH adjusting agent
Kind is several；
The melanin precursor includes dopachrome, 5,6- dihydroxy indole, one in 5,6- dihydroxy indole -2- carboxylic acid
Kind is several.
Preferably, the wetting and penetrating agent is one or both of L-cysteine, glutathione；
The dyeing auxiliaries is the metallic salt containing divalent or Tricationic；
The thickener is sodium carboxymethylcellulose, carboxymethyl cellulose, carboxyethyl cellulose, starch, Silica hydrogel, bright
Glue, sodium alginate, guar gum, agar, soybean protein glue, polyethylene glycol, Sodium Polyacrylate, polyacrylamide, poly- γ-paddy ammonia
One or more of acid, hyaluronic acid sodium, polyacrylic acid, low-molecular polyethylene wax, xanthan gum；
The surfactant is glycerin monostearate, hexadecyltrimethylammonium chloride, cetanol, lauryl sulfonation
One or more of monomester succinate disodium, sulfosuccinic acid monoesters disodium, love Ma cream；
The moisturizer is one or more of glycerol, sorbierite, mannitol, ethylene glycol, glycerine, edible oil；
The pH adjusting agent is one or more of lysine, arginine, sodium bicarbonate.
Preferably, the divalent or Tricationic are Cu2+、Mg2+、Ca2+、Fe2+、Fe3+One or more of.
Preferably, in the A agent:
The weight percent that the wetting and penetrating agent accounts for A agent is 1%~10%；
The weight percent that the thickener accounts for A agent is 0.2%~2%；
The weight percent that the surfactant accounts for A agent is 0.1%~2%；
The weight percent that the moisturizer accounts for A agent is 1%~15%；
The weight percent that the pH adjusting agent accounts for hair dye is 1%~10%；
In the B agent:
The weight percent that the dyeing auxiliaries accounts for B agent is 0.5%~10%；
The weight percent that the thickener accounts for B agent is 0.2%~2%；
The weight percent that the surfactant accounts for B agent is 0.1%~2%；
The weight percent that the pH adjusting agent accounts for B agent is 0.1%~1%；
In the C agent:
The weight percent that the wetting and penetrating agent accounts for C agent is 1%~10%；
The weight percent that the thickener accounts for C agent is 0.2%~2%；
The weight percent that the surfactant accounts for C agent is 0.1%~2%；
The weight percent that the moisturizer accounts for C agent is 1%~15%；
The weight percent that the pH adjusting agent accounts for C agent is 1%~10%.
Preferably, the melanin precursor preparation the following steps are included:
(1) prepare reaction substrate；
(2) complex enzyme needed for extracting synthesis of melanin precursor in edible mushroom；
(3) reaction substrate in step 1 is catalyzed using complex enzyme made from step 2, obtains target product；
(4) gained target product in step 3 is subjected to conversion processing, obtains melanin precursor；
The reaction substrate is one of tyrosine, DOPA, dopamine or a variety of；
The edible mushroom be one of Pleurotus eryngii, agaricus bisporus, brown mushroom, straw mushroom, oyster mushroom, mushroom, black fungus, agrocybe or
It is a variety of.
Preferably, the target product conversion processing one of performs the following operation or a variety of: boiling water heating, heating are same
When be evaporated under reduced pressure, electrolysis, addition decarboxylase.
Preferably, the substrate is l-tyrosine.
Preferably, the concentration of substrate is 1mM~10mM.
Preferably, the complex enzyme extracted in the step 2 carries out purifying and immobilization processing；The method of the purifying is
Ultrafiltration and/or ion-exchange chromatography.
Preferably, described when immobilizing processing to complex enzyme, fixed material is molecular sieve, diatomite, sodium alginate
Deng well known immobilization material.
The application method of three dosage form melanin precursor substance hair dyes of the invention, the application method the following steps are included:
The A agent of above-mentioned three dosage forms melanin precursor substance hair dye will be uniformly applied to hair surface after dyeing hair cleaning, 15~
It is cleaned after 25min with clear water；B agent is uniformly applied to hair surface again, is cleaned after 15~25min with clear water；Then C agent is uniform
It is applied to hair surface, is cleaned after 15~25min with clear water, i.e., hair is dyed into black.
Preferably, the A agent, B agent, C agent usage ratio be 3:1:3~1:1:1, the ratio be mass ratio.
Melanin precursor substance hair dye of the present invention and its application method difference from prior art are:
1. the present invention synthesizes DOPA color by bioconversion by extracting the complex enzyme liquid in natural edible bacterium, by amino acid
Element, 5,6- dihydroxy indole and 5, the melanin precursors substance such as 6- dihydroxy indole -2- carboxylic acid, is main with these precursor substances
Hair dye ingredient, addition auxiliary material are prepared into the biotechnology of the multi-form biology melanin hair dye of Non-oxidized.This product avoids
The chemical hair dye ingredient such as p-PDA, quinhydrones, resorcinol has been used, these substances are proved to the carcinogenesis for having certain, and
And the allergic reaction of certain customers can be caused and do not include lead, mercury, arsenic etc. to the heavy metal of the toxic effect of human body at
Point；Avoid having used common strong oxidizer hydrogen peroxide, the ammonia with strong and stimulating smell in other chemical hair dyes simultaneously
The ingredients such as water alleviate the injury to hair and skin, while this product also has anti-ultraviolet function, and containing there are many amino
Acid is conducive to the health of scalp and the protection of hair.
2. the melanin precursor substance of hair dye small molecular of the invention enters to the end under the action of wetting and penetrating agent
In hair, the melanin precursor substance in hair is aoxidized using the oxygen in air, aggregates into macromolecular melanin granule, thus
It remains in hair, makes white hair that nature black be presented.Product of the present invention no sensitization non-stimulated to skin, it is highly-safe,
No pollution to the environment, and melanin precursor ingredient also has anti-ultraviolet function, plays a protective role to scalp and hair.
3. hair dye of the invention during production and use have raw material be easy to get, simple process, using effect can
By and it is safe and non-toxic the advantages that, be easy to the factorial production, using the enzyme solution extracted in edible mushroom, before being biologically converted into melanin
Body substance, and it is configured to the method for hair dye and production technology is not reported at home and abroad.Hair dye preparation method of the invention
There is good development prospect because having the above advantages.
4. hair dye of the invention has a variety of dosage forms it can be selected that according to the difference of two-form and three dosage forms, it is corresponding with
Different application methods, easy to use and flexible.
Melanin precursor substance hair dye of the invention and its application method are described further with reference to the accompanying drawing.
Detailed description of the invention
Fig. 1 is eumelanin synthetic reaction figure；
Fig. 2 is melanin precursor thin-layer chromatography composition measurement figure；
Fig. 3 is the abosrption spectrogram of 5,6- dihydroxy indole standard items；
Fig. 4 is the abosrption spectrogram of the obtained melanin precursor object solution of the present invention；
Fig. 5 is that the hoary-headed of people visually observes figure；
Fig. 6 is the hoary-headed micro- sem observation figure of people；
Fig. 7 is that the natural dark hair of people visually observes figure；
Fig. 8 is the micro- sem observation figure of the natural dark hair of people；
Fig. 9 visually observes figure for the commercially available dyeing hair of hair dye containing p-phenylenediamine；
Figure 10 for the commercially available dyeing hair of hair dye containing p-phenylenediamine micro- sem observation figure；
Figure 11 visually observes figure for hair dye institute of the present invention dyeing hair；
Figure 12 for hair dye institute of the present invention dyeing hair micro- sem observation figure；
Figure 13 is the effect picture that hair dye of the present invention is used for white yak wool dyeing；
Figure 14 is the practical hair dyeing effect figure of hair dye of the present invention
Scheme compared with Figure 15 hair dye of the present invention is anti-ultraviolet with commercially available hair dye.
Specific embodiment
Theory of the invention is to simulate the generating process (as shown in Figure 1) of eumelanin in human body dark hair, utilizes biological method
Synthesis of melanin precursor substance, and enter melanin precursor substance in hair by auxiliary material, then aggregate into melanin
And stay in hair, form permanent type hair dye.
The present invention is with tyrosine or derivatives thereof for substrate, obtain in enzymatic reaction using the complex enzyme liquid of extraction
Melanin precursor substance, including dopachrome, 5,6- dihydroxy indole, 5,6- dihydroxy indole -2- carboxylic acid etc., indoles -5,6
Quinone, indoles -5,6 quinone -2- carboxylic acid etc., wherein it is preferred that 5,6- dihydroxy indoles.In addition, the melanin precursor in the present invention does not limit to
It also may include the oligomer of diploid or more in monomer.
Invention is with tyrosine or derivatives thereof for substrate, is carried out obtained in enzymatic reaction using the complex enzyme liquid of extraction
Melanin precursor substance, including dopachrome, 5,6- dihydroxy indole, 5,6- dihydroxy indole -2- carboxylic acid etc., indoles -5,6
Quinone, indoles -5,6 quinone -2- carboxylic acid etc., wherein it is preferred that 5,6- dihydroxy indoles.In addition, the melanin precursor in the present invention does not limit to
It also may include the oligomer of diploid or more in monomer.The preparation method of melanin precursor is as follows.
Production process
Raw material: as substrate, SILVER REAGENT or food-grade tyrosine and its derivative, such as D or L-type junket ammonia can be used
Sour, D or L-type DOPA, dopamine etc., can be one such or several combinations, are also possible to the isomers of above-mentioned substance.
Enzyme solution: extracting complex enzyme liquid using breaking method from edible mushroom, and phosphoric acid, lemon can be added in the process of extraction
Lemon acid, Tris-HCl buffer, pH value are controlled 6.0~8.0.Crude enzyme liquid is obtained by modes such as centrifugations after extraction.
Crude enzyme liquid mentioned above is used directly for the synthetic reaction of product, can also further be handed over by ultrafiltration and ion
Column chromatography, the enzyme that the processing such as ammonium sulfate precipitation and/or tannic acid precipitating are purified are changed, cryo-conservation after freeze-drying dehydration can be grown
Phase keeps enzymatic activity.
Enzyme improves the activity of enzyme before use, can be coordinated processing or acid solution processing etc. by divalent copper ion.
Enzyme solution mentioned above, which can be added directly into reaction solution, carries out enzymatic reaction, in addition, from the stabilization for improving enzyme
Property and reuse efficiency, the easy degree of decomposition after use, avoid other oroteins from being mixed into reaction system etc. setting out,
The enzyme of immobilization can be used.Enzyme immobilizatio method is not particularly limited, and can pass through molecular sieve, diatomite, sodium alginate etc.
Well known process for fixation, the method that the affine absorption such as edible mushroom powder and hair powder can also be used are fixed.For solid
Determine the different different with the purifying requirement of enzyme to thick enzyme of method.
Reaction step:
(1) pretreatment of tyrosine substrate solution
When using l-tyrosine as substrate, for concentration of substrate range of choice in 1mM~10mM, tyrosine substrate solubility is smaller,
The means such as adjusting pH can be taken to increase its solubility.
(2) the bioconversion reaction of tyrosinase
In the bioreactor, it is previously added the l-tyrosine substrate solution of 1mM~10mM, is 1/ using according to volume ratio
5~1/30 ratio is added the complex enzyme liquid that previous step is extracted, then passes to air, keep air pressure 0.05Mpa~
0.2Mpa, air mass flow are 5LPM~20LPM, carry out bioconversion reaction, and timing sampling detects.
(3) it detects
Sample centrifugation is taken out, supernatant is taken to dilute 10 times, 300nm (5,6- dihydroxy are measured in ultraviolet specrophotometer
The absorption peak of indoles) and 475nm (absorption peak of dopachrome) absorbance.When 300nm absorbance be greater than 2.0, and
When the absorbance of 475nm is greater than 0.5, bioconversion reaction is terminated, is further processed.
Postprocessing working procedures:
(1) collection of melanin precursor substance
In above-mentioned conversion reaction solution, in addition to containing the melanin precursor substance generated, also containing some including remaining
Melanin after resolvase or the polymerization of immobilized enzyme and melanin precursor.Make to collect the melanin precursor object in the reaction solution
For the dyestuff of hair dyeing, therefore need to remove the melanin after remaining enzyme and polymerization.The side such as centrifugation, ultrafiltration can be passed through for enzyme
Method removes, and melanin after polymerization can be removed by the methods of adsorbing, filtering.The enzyme of removal may return to reaction process again
It utilizes, the melanin of removal can be applied to other products.It finally collects obtained solution and predominantly contains melanin precursor object
Matter solution, including dopachrome, 5,6- dihydroxy indole, 5,6- dihydroxy indole are full, 5,6- dihydroxy indole -2- carboxylic acid etc.,
Wherein dopachrome accounts for large percentage.
(2) adjustment of melanin precursor object composition
In melanin precursor object solution, white hair can be made to contaminate black most important hair dyeing effective component 5,6- dihydroxy
Indoles is penetrated into inside hair by wetting, and aggregates into melanin granule in hair using the oxygen in air,
It is left in hair cortex, makes hair that black be presented.According to this final purpose, the composition to melanin precursor substance solution is needed
Carry out conversion adjustment.It can be heated by (I) boiling water, (II) heats while depressurizing, (III) is electrolysed, (IV) is using in decarboxylase
One or more processing is to adjust portfolio ratio, so that melanin precursor object based on dopachrome is to 5,6- dihydroxy
The conversion of base indoles.Obtained melanin precursor object solution is for preparing hair dye.
(3) measurement of melanin precursor object ingredient
By melanin precursor object solution obtained by the above method, thin-layer chromatography can be selected to carry out Major Components
(Fig. 2), while detection 5, the content (Fig. 3, Fig. 4) of 6- dihydroxy indole are carried out in wavelength 300nm with ultraviolet spectrophotometry.
(4) preparation method of hair dye
Ingredient in hair dye of the invention mainly includes melanin precursor substance, wetting and penetrating agent, thickener, surface work
Property agent, pH adjusting agent, dyeing auxiliaries, moisturizer, can also be added some nutritional ingredients, such as some plant extracts, Chinese medicine
Ingredient etc..As the main component melanin precursor substance of hair dye, content preferably 1%~5%；Wetting and penetrating agent (L- half
One or both of cystine, glutathione)；Dyeing auxiliaries (one in the metal salts such as Cu2+, Mg2+, Ca2+, Fe2+, Fe3+
Kind or more than one)；Thickener is (sodium carboxymethylcellulose, carboxymethyl cellulose, carboxyethyl cellulose, starch, Silica hydrogel, bright
Glue, sodium alginate, guar gum, agar, soybean protein glue, polyethylene glycol, Sodium Polyacrylate, polyacrylamide, poly- γ-base paddy ammonia
Acid, one of hyaluronic acid sodium, low-molecular polyethylene wax, xanthan gum etc. or more than one)；Surfactant (monostearate
Glyceride, hexadecyltrimethylammonium chloride, cetanol, lauryl sulfosuccinic acid monoester disodium, sulfosuccinic acid monoesters two
Sodium, one or more of love Ma cream)；Moisturizer (glycerol, sorbierite, mannitol, ethylene glycol, glycerine, one in edible oil
Kind or more than one)；PH adjusting agent (at least one of lysine, arginine, sodium bicarbonate).
Embodiment 1
The preparation method of the present embodiment melanin precursor sequentially includes the following steps:
1. the extraction and measurement of tyrosinase
It takes the distilled water of 500 grams of the broken material of Pleurotus eryngii addition 1000ml to extract tyrosine enzyme solution or utilizes 1000ml
Neutral phosphate buffer extracts tyrosine enzyme solution.
The measuring method of tyrosine enzyme activity is as follows: taking the substrate solution of 0.1ml mushroom complex enzyme liquid and 3.9ml in 317nm
10min is reacted, the definition of enzyme activity unit U is that every 1min causes the variation of 0.01 unit of absorbance to be 1 enzyme activity unit.
When the result of measurement is greater than 1000U, this enzyme solution can be used for melanin precursor accumulation reaction below.
2. the accumulation of melanin precursor is reacted
Compound concentration is the tyrosine solution 2L of 1mM-10mM, and it is raw for 10L that content volume is added in tyrosine substrate solution
In object reactor, using temperature control system, temperature is controlled at 30 DEG C, according to 1/20 ratio of substrate volume, in addition
The complex enzyme liquid of onestep extraction then passes to air, and holding air pressure is 0.10Mpa, and air mass flow 10LPM is given birth to
Object conversion reaction, and timing sampling detects.
The fluid sample of taking-up is centrifuged, after taking supernatant to dilute 10 times, measured in ultraviolet specrophotometer 300nm and
The absorbance of 475nm.When absorbance of the absorbance of 300nm greater than 2.0 and 475nm is greater than 0.5, it is anti-to terminate bioconversion
It answers, is further processed.
3. the collection of melanin precursor object
Melanin precursor object solution obtained above is centrifuged off tyrosinase and foreign protein in 8000rpm；Supernatant adds
Enter 5% active carbon, the melanin granule polymerizeing during adsorption reaction, with more in the melanin precursor object solution obtained at this time
Based on bar pigment.
4. the adjustment of melanin precursor composition
5,6- dihydroxy indole is converted to by dopachrome, using Rotary Evaporators by operating melanin precursor solution
Adjustment is collected.
Embodiment 2
The preparation method of the present embodiment melanin precursor sequentially includes the following steps:
1. the extraction of tyrosinase
Brown mushroom is crushed using juice extractor, the lemon acid buffering that 1000ml pH value is 6.8 is added in 500g fructification disintegrating slag
Liquid extracts enzyme solution, spare using being centrifuged after 5% activated carbon adsorption processing.
2. the fixation of tyrosinase
The diatomite of 200 mesh is passed through into 60 DEG C of heat treatment 30min, impregnates 20min, centrifugation by perphosphate buffer
Supernatant is removed, the diatomite obtained after processing is added in above-mentioned enzyme solution and fixes 5h, it is spare to obtain immobilised enzymes.
3. the accumulation of melanin precursor is reacted
Compound concentration is the tyrosine solution 2L of 5mM, and the method that can take heating for dissolving makes it completely dissolved.
It is in 10L bioreactor, using temperature control system, by temperature that content volume, which is added, in tyrosine substrate solution
Control is added 100 grams of previous step immobilised enzymes at 30 DEG C, then passes to air, and holding air pressure is 0.10Mpa, air stream
Amount is 10LPM, carries out bioconversion reaction, and timing sampling detects.
The fluid sample of taking-up by centrifugation after, supernatant dilute 10 times after, in ultraviolet specrophotometer measure 300nm with
And the absorbance of 475nm.When the absorbance of 300nm is greater than 2.0, and the absorbance of 475nm is greater than 0.5, terminates biology and turn
Change reaction, is further processed.
4. the collection of melanin precursor
Melanin precursor solution obtained above removes immobilised enzymes and foreign protein in 12000rpm, 4 DEG C of centrifugation 5min；
In the melanin precursor solution obtained at this time based on dopachrome.
5. the adjustment of melanin precursor composition
Adjustment process is the same as embodiment 1.
Embodiment 3
The preparation method of the present embodiment melanin precursor sequentially includes the following steps:
1. the extraction of tyrosinase
Agaricus bisporus is crushed using juice extractor, 500g disintegrating slag is added the Tris- hydrochloride buffer that 1000mlpH value is 7.0 and mentions
Enzyme solution is taken, supernatant is extracted in centrifugation, polysaccharide and foreign protein etc. in enzyme solution is adsorbed using 10% macroporous absorbent resin, treated
Enzyme solution is spare under conditions of placing 4 DEG C.
2. the accumulation of melanin precursor is reacted
Compound concentration is the tyrosine solution 2L of 10mM, and the method that can take heating for dissolving makes it completely dissolved.
Reaction process is the same as embodiment 1.
The fluid sample of taking-up is filtered through filter paper, and after filtrate dilutes 10 times, 300nm is measured in ultraviolet specrophotometer
And the absorbance of 475nm.When the absorbance of 300nm is greater than 2.0, and the absorbance of 475nm is greater than 0.5, biology is terminated
Conversion reaction, acquired solution contain primary dye ingredient -5,6 dihydroindole of hair dyeing.
Embodiment 4
The two dosage form melanin precursor substance hair dyes of the present embodiment are prepared according to the following steps: will be made in embodiment 1-3
Melanin precursor substance and hair dye usual auxiliaries and water, paste or watery dye are respectively prepared under conditions of depressurizing nitrogen charging
Agent A agent and B agent are sent out, wherein the selection of specific raw material and proportion are as shown in Table 1 and Table 2, wherein raw materials used is commercially available.
In 1 embodiment 4 of table in A agent each raw material type and amount
Wherein, melanin precursor substance is obtained in above-described embodiment 1-3, and above-mentioned percentage is the weight for accounting for hair dye
Percentage is measured, surplus is water.
In 2 embodiment 4 of table in B agent each raw material type and amount
Above-mentioned percentage is the weight percent for accounting for hair dye, and surplus is water.
Embodiment 5
The application method of the melanin precursor substance hair dye of the present embodiment sequentially includes the following steps:
It will be equal by the A agent of two dosage form melanin precursor substance hair dyes obtained in embodiment 4 after dyeing hair cleaning
It is even to be applied to hair surface, it is cleaned after 30min with clear water；B agent is uniformly applied to hair surface again, after 25min, is cleaned with clear water,
Hair is dyed into black.It is once dyed i.e. visible color development significantly to black, from the effect contrast figure (Figure 14) of hair dyeing front and back
As can be seen that the hair color after the hair dyeing of this hair dye is uniform, and hair is smooth glossy.
Embodiment 6
The application method of the melanin precursor substance hair dye of the present embodiment and embodiment 5 the difference is that: hair dyeing
Residence time of the agent A agent on hair is 50min；Residence time of the hair dye B agent on hair is 15min；Once dyed
I.e. visible color development significantly blacks.
Embodiment 7
The application method of the melanin precursor substance hair dye of the present embodiment and embodiment 5 the difference is that: hair dyeing
Residence time of the agent on hair is 40min；Residence time of the hair dye A agent on hair is 20min；Once dyed
See that color development significantly blacks.
Embodiment 8
The three dosage form melanin precursor substance hair dyes of the present embodiment are prepared according to the following steps: will be made in embodiment 1-3
Melanin precursor substance and hair dye usual auxiliaries and water, paste or watery dye are respectively prepared under conditions of depressurizing nitrogen charging
Agent A agent, B agent and C agent are sent out, wherein the selection of specific raw material and proportion are as shown in Table 3-5, wherein raw materials used is commercially available.
In 3 embodiment 8 of table in A agent each raw material type and amount
Wherein, melanin precursor substance is obtained in above-described embodiment 1-3, and above-mentioned percentage is the weight for accounting for hair dye
Percentage is measured, surplus is water.
In 4 embodiment 8 of table in B agent each raw material type and amount
Above-mentioned percentage is the weight percent for accounting for hair dye, and surplus is water.
In 5 embodiment 8 of table in C agent each raw material type and amount
Wherein, melanin precursor substance is obtained in above-described embodiment 1-3, and above-mentioned percentage is the weight for accounting for hair dye
Percentage is measured, surplus is water.
Contrast verification test
Hair dye of the present invention is compared with commercially available hair dye
The white hair for taking equivalent uniformly applies after mixing the hair dye A agent of commercially available two-form with B agent after shampoo cleans
In on hair, loose colour is washed away with clear water after 40min.
The white hair for taking equivalent, after shampoo and clear water are washed, by hair dye of the invention according to the step pair in embodiment 5
Hair is dyed.
Then, white hair, the natural dark hair of people and the effect of the dark hair contaminated are observed under the microscope (see Fig. 5-Figure 12).
It can be seen that from result, this product is ideal and smaller to the damage of hair to the hair dyeing effect of white hair, and the hair after contaminating is more
Close to human body natural's dark hair.
Hair dye of the present invention is used for the dyeing of white yak hair
The structure of white yak hair is similar with human hair, is usually utilized to the substitute examined as hair dyeing effect.
The A agent of product made from embodiment 4 is applied on white yak hair with comb, both forward and reverse directions comb repeatedly, make cream
Body is evenly distributed on yak hair, is stopped 30min, is then cleaned with clear water；B agent is uniformly applied to hair surface, 25min again
Afterwards, it is cleaned with clear water, is wiped with towel to not dripping.
From the yak hair color that can be seen that in the effect contrast figure (Figure 13) of hair dyeing front and back after the hair dyeing of this hair dye
It is uniformly pitch-black.
This hair dye and commercially available hair dye ultraviolet absorption spectrum compare
The independent A agent of hair dye of the present invention is accurately weighed respectively, and object, the A agent of three dosage forms, B are closed in A agent, the B agent of two-form very much
Agent, each 8mg of C agent composition；It is denoted as No. 1 sample, No. 2 samples and No. 3 samples；It is control with commercially available L'Oreal's hair dye,
Uniformly it is applied to 4cm23M medical adhesive tape on, in ultraviolet specrophotometer scan 280nm~400nm absorbance, obtain
As a result as shown in figure 15.No. 2 samples and No. 3 samples of product of the present invention, are wanted in the absorbance of UVB wave band (280nm~310nm)
It is apparently higher than commercially available hair dye.
Although specific embodiments of the present invention have been described above, it will be appreciated by those of skill in the art that these
It is merely illustrative of, protection scope of the present invention is defined by the appended claims.Those skilled in the art is not carrying on the back
Under the premise of from the principle and substance of the present invention, various changes or modifications can be made to these embodiments, but these are changed
Protection scope of the present invention is each fallen with modification.
Claims (8)
1. a kind of preparation method of two dosage form melanin precursor substance hair dyes, which is characterized in that
The preparation of the hair dye includes following process:
(a) edible mushroom complex enzyme liquid is extracted, purified, fixed and saved；
(b) reaction substrate is subjected to biotransformation, obtains melanin precursor；
(c) reaction solution obtained above is adjusted, improves the concentration of 5,6- dihydroxy indole；
(d) melanin precursor substance obtained above is mixed with auxiliary material, two component hair dyes can be obtained；
The preparation of the melanin precursor the following steps are included:
(1) prepare reaction substrate；
(2) complex enzyme needed for extracting synthesis of melanin precursor in edible mushroom；
(3) reaction substrate in step (1) is catalyzed using complex enzyme made from step (2), obtains target product；
(4) gained target product in step (3) is subjected to conversion processing, obtains melanin precursor object；
The reaction substrate is one of tyrosine, DOPA, dopamine or a variety of；
The edible mushroom is one of Pleurotus eryngii, agaricus bisporus, brown mushroom, straw mushroom, oyster mushroom, mushroom, black fungus, agrocybe or more
Kind；
The hair dye includes: A agent and B agent；
The A agent includes melanin precursor, auxiliary agent A and water；The auxiliary agent A be wetting and penetrating agent, thickener, surfactant,
One or more of moisturizer, pH adjusting agent；
The melanin precursor include one of dopachrome, 5,6- dihydroxy indole, 5,6- dihydroxy indole -2- carboxylic acid or
It is several；
The weight percent that the melanin precursor accounts for A agent is 0.01%~5%；
The B agent includes auxiliary agent B and water；The auxiliary agent B is dyeing auxiliaries, thickener, surfactant, one in pH adjusting agent
Kind is several.
2. the preparation method of two dosage forms melanin precursor substance hair dye according to claim 1, it is characterised in that:
The wetting and penetrating agent is one or both of L-cysteine, glutathione；
The dyeing auxiliaries is the metallic salt containing divalent or Tricationic；
The thickener is sodium carboxymethylcellulose, carboxymethyl cellulose, carboxyethyl cellulose, starch, Silica hydrogel, gelatin, sea
Mosanom, guar gum, agar, soybean protein glue, polyethylene glycol, Sodium Polyacrylate, polyacrylamide, Polyurethane-epoxy resin, glass
One or more of matter acid sodium, polyacrylic acid, low-molecular polyethylene wax, xanthan gum；
The surfactant is glycerin monostearate, hexadecyltrimethylammonium chloride, cetanol, lauryl sulfosuccinate
One or more of acid monoester disodium, sulfosuccinic acid monoesters disodium, love Ma cream；
The moisturizer is one or more of glycerol, sorbierite, mannitol, ethylene glycol, glycerine, edible oil；
The pH adjusting agent is one or more of lysine, arginine, sodium bicarbonate.
3. the preparation method of two dosage forms melanin precursor substance hair dye according to claim 1 or 2, it is characterised in that:
In the A agent:
The weight percent that the wetting and penetrating agent accounts for A agent is 1%~10%；
The weight percent that the thickener accounts for A agent is 0.2%~2%；
The weight percent that the surfactant accounts for A agent is 0.1%~2%；
The weight percent that the moisturizer accounts for A agent is 1%~15%；
The weight percent that the pH adjusting agent accounts for hair dye is 1%~10%；
In the B agent:
The weight percent that the dyeing auxiliaries accounts for B agent is 0.5%~10%；
The weight percent that the thickener accounts for B agent is 0.2%~2%；
The weight percent that the surfactant accounts for B agent is 0.1%~2%；
The weight percent that the pH adjusting agent accounts for B agent is 0.1%~1%.
4. the application method of two dosage form melanin precursor substance hair dyes described in a kind of claim 1, which is characterized in that described to make
The following steps are included: it will be after dyeing hair cleans, by two dosage form melanin precursor substance hair dye described in claim 1 with method
A agent be uniformly applied to hair surface, cleaned after 30~50min with clear water；B agent is uniformly applied to hair surface, 15~25min again
Afterwards, it is cleaned with clear water, i.e., hair is dyed into black.
5. a kind of preparation method of three dosage form melanin precursor substance hair dyes, which is characterized in that
The preparation of the hair dye includes following process:
(a) edible mushroom complex enzyme liquid is extracted, purified, fixed and saved；
(b) reaction substrate is subjected to biotransformation, obtains melanin precursor；
(c) reaction solution obtained above is handled, improves the concentration of 5,6- dihydroxy indole；
(d) melanin precursor substance obtained by the above method is mixed with auxiliary material, three component hair dyes can be obtained；
The preparation of the melanin precursor the following steps are included:
(1) prepare reaction substrate；
(2) complex enzyme needed for extracting synthesis of melanin precursor in edible mushroom；
(3) reaction substrate in step (1) is catalyzed using complex enzyme made from step (2), obtains target product；
(4) gained target product in step (3) is subjected to conversion processing, obtains melanin precursor；
The reaction substrate is one of tyrosine, DOPA, dopamine or a variety of；
The edible mushroom is one of Pleurotus eryngii, agaricus bisporus, brown mushroom, straw mushroom, oyster mushroom, mushroom, black fungus, agrocybe or more
Kind；
The hair dye includes: A agent, B agent and C agent；
The A agent includes melanin precursor, auxiliary agent A and water；Melanin precursor accounts for the weight percent of A agent and is in the A agent
0.01%~5%；The auxiliary agent A is one of wetting and penetrating agent, thickener, surfactant, moisturizer, pH adjusting agent
Or it is several；
The B agent includes auxiliary agent B and water；The auxiliary agent B is dyeing auxiliaries, thickener, surfactant, one in pH adjusting agent
Kind is several.
The C agent includes melanin precursor, auxiliary agent C and water；Melanin precursor accounts for the weight percent of C agent and is in the C agent
0.01%~5%；The auxiliary agent C is one of wetting and penetrating agent, thickener, surfactant, moisturizer, pH adjusting agent
Or it is several；
The melanin precursor include one of dopachrome, 5,6- dihydroxy indole, 5,6- dihydroxy indole -2- carboxylic acid or
It is several.
6. the preparation method of three dosage forms melanin precursor substance hair dye according to claim 5, it is characterised in that:
The wetting and penetrating agent is one or both of L-cysteine, glutathione；
The dyeing auxiliaries is the metallic salt containing divalent or Tricationic；
The thickener is sodium carboxymethylcellulose, carboxymethyl cellulose, carboxyethyl cellulose, starch, Silica hydrogel, gelatin, sea
Mosanom, guar gum, agar, soybean protein glue, polyethylene glycol, Sodium Polyacrylate, polyacrylamide, Polyurethane-epoxy resin, glass
One or more of matter acid sodium, polyacrylic acid, low-molecular polyethylene wax, xanthan gum；
The surfactant is glycerin monostearate, hexadecyltrimethylammonium chloride, cetanol, lauryl sulfosuccinate
One or more of acid monoester disodium, sulfosuccinic acid monoesters disodium, love Ma cream；
The moisturizer is one or more of glycerol, sorbierite, mannitol, ethylene glycol, glycerine, edible oil；
The pH adjusting agent is one or more of lysine, arginine, sodium bicarbonate.
7. the preparation method of two dosage forms melanin precursor substance hair dye according to claim 5 or 6, it is characterised in that:
In the A agent:
The weight percent that the wetting and penetrating agent accounts for A agent is 1%~10%；
The weight percent that the thickener accounts for A agent is 0.2%~2%；
The weight percent that the surfactant accounts for A agent is 0.1%~2%；
The weight percent that the moisturizer accounts for A agent is 1%~15%；
The weight percent that the pH adjusting agent accounts for hair dye is 1%~10%；
In the B agent:
The weight percent that the dyeing auxiliaries accounts for B agent is 0.5%~10%；
The weight percent that the thickener accounts for B agent is 0.2%~2%；
The weight percent that the surfactant accounts for B agent is 0.1%~2%；
The weight percent that the pH adjusting agent accounts for B agent is 0.1%~1%；
In the C agent:
The weight percent that the wetting and penetrating agent accounts for C agent is 1%~10%；
The weight percent that the thickener accounts for C agent is 0.2%~2%；
The weight percent that the surfactant accounts for C agent is 0.1%~2%；
The weight percent that the moisturizer accounts for C agent is 1%~15%；
The weight percent that the pH adjusting agent accounts for C agent is 1%~10%.
8. a kind of application method of three dosage form melanin precursor substance hair dyes, which is characterized in that the application method include with
Lower step: will be uniform by the A agent of three dosage form melanin precursor substance hair dyes described in claim 5 after dyeing hair cleaning
It is applied to hair surface, is cleaned after 15~25min with clear water；B agent is uniformly applied to hair surface again, uses clear water after 15~25min
Cleaning；Then C agent is uniformly applied to hair surface, is cleaned after 15~25min with clear water, i.e., hair is dyed into black.
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| CN113197784A (en) * | 2021-04-13 | 2021-08-03 | 北京源天彩生物科技有限公司 | Preparation method and application of hair dyeing product containing melanin precursor prepared by biological fermentation |
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| CN116115512B (en) * | 2023-04-17 | 2023-08-04 | 知想(南京)生物科技有限公司 | Oligomeric melanin inclusion, and preparation method and application thereof |
| WO2024027859A1 (en) * | 2023-09-11 | 2024-02-08 | 北京芙洛迪科技有限公司 | Double-agent-form hair dye and use method therefor |
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