阅读说明：本技术 黑色素前体物多巴醌的制备方法及应用 (Preparation method and application of melanin precursor dopaquinone ) 是由 张天 杜成强 于 2020-07-20 设计创作，主要内容包括：本发明公开了一种黑色素前体物多巴醌的制备方法及应用,克服了传统染发剂中染发成分对苯二胺具有毒性的缺点。本发明的技术方案为：以黧豆提取物左旋多巴为主要生色活性物质,使用土豆皮提取的酪氨酸酶在生物工艺催化反应下生成黑色素前体多巴醌。本发明所制得的黑色素前体物多巴醌,替代了传统染发剂中染发成分对苯二胺,将其应用于染发剂中,使用植物提取物进行制备,制备过程对环境友好,安全环保,所制备得到的黑色素前体物多巴醌对人体没有毒性。(The invention discloses a preparation method and application of a melanin precursor dopaquinone, which overcome the defect that a hair dyeing component p-phenylenediamine in a traditional hair dye has toxicity. The technical scheme of the invention is as follows: largyrum pruriens extract levodopa is used as a main chromogenic active substance, and tyrosinase extracted from potato peel is used for generating dopaquinone precursor of melanin under the catalysis reaction of a biological process. The prepared melanin precursor dopaquinone replaces a hair dyeing component p-phenylenediamine in the traditional hair dye, is applied to the hair dye, is prepared by using the plant extract, is environment-friendly and safe and environment-friendly in the preparation process, and has no toxicity to a human body.)

1. A preparation method of a melanin precursor dopaquinone is characterized by comprising the following steps:

largyrum pruriens extract levodopa is used as a main chromogenic active substance, and tyrosinase extracted from potato peel is used for generating dopaquinone precursor of melanin under the catalysis reaction of a biological process.

2. The method for preparing dopaquinone as a melanin precursor according to claim 1, wherein the method comprises the following steps:

the method comprises the following specific steps:

step one, preparing a mucuna pruriens extract:

crushing chenopodium album, extracting with a mixed solution of 30% ethanol and 0.1% acetic acid at normal temperature, and filtering to obtain an extract; concentrating the extracting solution under reduced pressure, separating out crystals, standing overnight, and filtering to obtain a levodopa crude product; dissolving the crude product, filtering, adding vitamin C into the filtrate, adjusting pH to 3.5, precipitating crystal, standing for 4 hr, and filtering; washing and drying the filter cake to obtain a mucuna pruriens extract;

step two, preparing tyrosine crude enzyme freeze-dried powder:

standing potato tuber overnight, thawing with clear water to obtain pulverized skin, adding polyvinylpyrrolidone and phosphoric acid buffer solution, homogenizing, filtering, slowly adding ammonium sulfate powder, and centrifuging; taking supernatant, continuously adding ammonium sulfate, and centrifuging; carrying out vacuum freeze drying on the precipitate to obtain tyrosine crude enzyme freeze-dried powder;

step three, performing biocatalytic reaction by using the extracts prepared in the step one and the step two:

preparing a lathy bean extract solution with the mass fraction of 10%, adding the lathy bean extract solution into a bioreactor, controlling the temperature to be 35 ℃, adding a tyrosine crude enzyme freeze-dried powder solution with the mass fraction of 30% according to the proportion of 1/10 of the volume of the lathy bean extract solution, carrying out biotransformation reaction, carrying out timing sampling detection, and stopping the biotransformation reaction when the absorbance at 300nm is more than 2.0 and the absorbance at 475nm is more than 0.5 to obtain a crude melanin precursor;

step four, crude product purification:

and (4) performing centrifugal deproteinization, activated carbon adsorption and melanin particle removal, ethyl acetate extraction and rotary evaporation solvent removal purification treatment on the melanin precursor crude product obtained in the step three to obtain the melanin precursor dopaquinone.

3. The method for preparing dopaquinone as a melanin precursor according to claim 1, wherein the method comprises the following steps:

in the first step, the standing temperature is 0-10 ℃; the drying temperature is 60-70 ℃.

4. Use of dopaquinone as melanin precursor in hair dyeing prepared by the preparation method of claims 1-3.

Technical Field

The invention belongs to the field of special cosmetics, and particularly relates to a preparation method and application of a melanin precursor dopaquinone.

Background

Modern chemical hair dyes have started from the discovery and use of p-phenylenediamine, which has been optimized and improved for many years, but the toxicity and side effects of the substances and derivatives thereof cannot be avoided. The reason is that PPD (p-phenylenediamine) and PTD (2, 5-diaminotoluene) which are key raw materials are main allergens causing allergy, have strong infiltration capacity and can be quickly diffused into the body through hair follicles in the process of dyeing hair, thereby bringing health hidden troubles and uncomfortable feeling. The national regulations stipulate that the maximum permissible concentrations of PPD and PTD in hair dyes are 6% and 10%, respectively, while the European maximum concentrations are 2% and 4%. According to statistics, the sensitization rate of PPD is 0.1% -l%, and the sensitization rate of PPD in the patch test of allergic patients is 2% -5%. PPD is more common for hair practitioners with contact dermatitis occurring at 19% to 35%. In addition, the hair dye also contains other unsafe auxiliary agents: the resorcinol can stimulate skin and mucosa, and can be rapidly absorbed through skin to cause poisoning symptoms; the p-aminophenol, the o-aminophenol and the m-aminophenol have double toxicity of aniline and phenol, and cause methemoglobin and asthma through skin absorption; thioglycolic acid has strong irritation, can cause serious damage when contacting with eyes, has sensitization effect on skin, and can cause dermatitis. Therefore, the hair dye is high in efficiency, but is not safe, and unpleasant odor can be emitted in the hair dyeing process, so that the hair dyeing experience is seriously influenced.

The plant hair dye is characterized in that natural dyes and surfactants with positive charges are combined into particles with small particle sizes and attached to the surface of hair, a plurality of plant dyes can be mixed to present different colors, or the plant dyes and water-soluble pigments are matched to form different colors, the dye molecules only act with hair scales in an adsorption mode and are generally classified into temporary or semi-permanent hair dyes, and the plant sources mainly comprise plants with rich pigment contents, such as henna, glabellum vulgare, papaverine, crocus sativus, curcumin and the like. However, the pure plant hair dye including henna powder also contains PPD substances, and is substantially characterized in that a part of plant extracts are added into the p-phenylenediamine hair dye, wherein effective dyeing components are p-phenylenediamine substances which are toxic and harmful to the body.

Disclosure of Invention

The invention aims to provide a preparation method and application of a melanin precursor dopaquinone.

In order to achieve the purpose, the invention provides a preparation method of a melanin precursor dopaquinone, which takes Lavida pruriens extract levodopa as a main chromogenic active substance, and tyrosinase extracted from potato peel is used for generating the melanin precursor dopaquinone under the catalysis reaction of a biological process.

A preparation method of a melanin precursor dopaquinone comprises the following specific steps:

step one, preparing a mucuna pruriens extract:

pulverizing semen Sojae Atricolor, extracting with mixed solution of 30% ethanol and 0.1% acetic acid at room temperature, and filtering to obtain extractive solution; concentrating the extracting solution under reduced pressure, separating out crystals, standing overnight, and filtering to obtain a levodopa crude product; dissolving the crude product, filtering, adding a small amount of vitamin C into the filtrate, adjusting pH to 3.5, precipitating crystals, standing for 4h, and filtering; washing and drying the filter cake to obtain a mucuna pruriens extract;

step two, preparing tyrosine crude enzyme freeze-dried powder:

standing potato tuber overnight, thawing with clear water to obtain pulverized skin, adding polyvinylpyrrolidone and phosphoric acid buffer solution, homogenizing, filtering, slowly adding ammonium sulfate powder, and centrifuging; taking supernatant, continuously adding ammonium sulfate, and centrifuging; carrying out vacuum freeze drying on the precipitate to obtain tyrosine crude enzyme freeze-dried powder;

step three, performing biocatalytic reaction by using the extracts prepared in the step one and the step two:

preparing a lathy bean extract solution with the concentration of 10%, adding the lathy bean extract solution into a bioreactor, controlling the temperature to be 35 ℃, adding a tyrosine crude enzyme freeze-dried powder solution with the mass fraction of 30% according to the proportion of 1/10 in the volume of the lathy bean extract solution, carrying out biotransformation reaction, carrying out timing sampling detection, and stopping the biotransformation reaction when the absorbance at 300nm is more than 2.0 and the absorbance at 475nm is more than 0.5 to obtain a crude melanin precursor;

and step four, carrying out centrifugal deproteinization, activated carbon adsorption and melanin particle removal, ethyl acetate extraction and rotary evaporation solvent removal purification treatment on the melanin precursor crude product obtained in the step three to obtain the melanin precursor dopaquinone.

The application of the melanin precursor dopaquinone prepared by the preparation method in the aspect of hair dye.

Compared with the prior art, the invention has the beneficial effects that:

the melanin precursor dopaquinone prepared by the invention replaces a hair dyeing component p-phenylenediamine in the traditional hair dye, is applied to the hair dye, is prepared by using the plant extract, is environment-friendly in preparation process, safe and environment-friendly, and has no toxicity to human bodies.

Drawings

FIG. 1 is a diagram showing the dyeing effect of the hair dye prepared from dopaquinone as a melanin precursor prepared in the example.

Detailed Description

The present invention will be described in further detail with reference to specific examples, but the embodiments of the present invention include, but are not limited to, the scope shown in the following examples.