Multi-dose melanin precursor substance hair dye and using method thereof
A technology of melanin precursors and hair dyes, which can be used in hair care, pharmaceutical formulations, cosmetic preparations, etc., and can solve problems such as environmental pollution, lack of pigment extraction technology, and long time consumption
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Embodiment 1
[0132] The preparation method of the melanin precursor of this embodiment is carried out according to the following steps:
[0133] 1. Extraction and determination of tyrosinase
[0134] Take 500 grams of the crushed Pleurotus eryngii and add 1000ml of distilled water to extract the tyrosinase solution or use 1000ml of neutral phosphate buffer to extract the tyrosinase solution.
[0135] The determination method of tyrosinase activity is as follows: take 0.1ml mushroom complex enzyme solution and 3.9ml substrate solution to react at 317nm for 10min, the enzyme activity unit U is defined as a change of 0.01 unit of absorbance per 1min as 1 enzyme activity unit. When the measured result is greater than 1000U, this enzyme solution can be used for the following melanin precursor accumulation reaction.
[0136] 2. Accumulation of melanin precursors
[0137] Prepare 2L of tyrosine solution with a concentration of 1mM-10mM, add the tyrosine substrate solution to the bioreactor with a volume ...
Embodiment 2
[0144] The preparation method of the melanin precursor of this embodiment is carried out according to the following steps:
[0145] 1. Extraction of Tyrosinase
[0146] The brown mushroom was crushed by a juice extractor, and 500 g of fruit body slag was added to 1000 ml of citric acid buffer solution with a pH of 6.8 to extract the enzyme solution, which was adsorbed by 5% activated carbon and centrifuged for use.
[0147] 2. Immobilization of Tyrosinase
[0148] Heat 200 mesh diatomaceous earth at 60°C for 30 minutes, then soak in phosphate buffer for 20 minutes, centrifuge to remove the supernatant, add the diatomaceous earth obtained after the treatment to the above enzyme solution and fix for 5 hours to obtain immobilized enzymes for use .
[0149] 3. Accumulation of melanin precursors
[0150] Prepare 2L of tyrosine solution with a concentration of 5mM, which can be completely dissolved by heating and dissolving.
[0151] Add the tyrosine substrate solution to a bioreactor with a v...
Embodiment 3
[0158] The preparation method of the melanin precursor of this embodiment is carried out according to the following steps:
[0159] 1. Extraction of Tyrosinase
[0160] Use a juice extractor to crush Agaricus bisporus, add 500g of the residue to 1000ml of Tris-hydrochloric acid buffer with a pH of 7.0 to extract the enzyme solution, centrifuge to extract the supernatant, and use 10% macroporous resin to adsorb polysaccharides and impurities in the enzyme solution. The treated enzyme solution is placed at 4°C for use.
[0161] 2. Accumulation of melanin precursors
[0162] Prepare 2L of tyrosine solution with a concentration of 10mM, which can be completely dissolved by heating and dissolving.
[0163] The reaction process is the same as in Example 1.
[0164] The liquid sample taken out was filtered through filter paper, and the filtrate was diluted 10 times, and the absorbance at 300 nm and 475 nm was measured in an ultraviolet spectrophotometer. When the absorbance at 300 nm is great...
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