Application of alginic acid/sodium alginate as tyrosinase inhibitor
A technology of tyrosinase and sodium alginate, which is applied in skin care preparations, cosmetic preparations, cosmetic preparations and other directions, can solve the problems of complex methods, difficult tyrosine inhibitors, high toxicity and the like, and achieves safety Application, inhibition of melanin hyperplasia, inhibition of TYR effect
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Embodiment 1
[0043] Example 1. Determination of TYR activity
[0044] The melanin produced by the catalytic substrate of TYR, L-dopa, can be used to investigate the change in the activity of TYR by measuring the change in absorbance value using an ultraviolet spectrophotometer. The specific operation method is as follows:
[0045] S1. Add 3 mL of PBS buffer with pH=6.8 to the test tube, and then add 2.8 mL of 1 mmol / L L-dopa solution to obtain L-dopa PBS buffer;
[0046] S2. Place the L-dopa PBS buffer prepared in step S1 in a 32°C water bath for 20 minutes, and then take 0.1 mL of 0.01 mg / mL TYR solution and immediately add it to the insulated L-dopa PBS buffer to mix;
[0047] S3. Then use an ultraviolet spectrophotometer to measure the absorbance of the reaction solution at 475nm, draw a curve of the change of TYR activity with unit time, and stipulate that the absorbance value of the reaction solution at 475nm wavelength increases by 0.001 per minute as a TYR activity unit (U / min).
[0048] f...
Embodiment 2
[0050] Example 2. Determination of TYR inhibition rate
[0051] The specific operation method of Example 2 is basically the same as that of Example 1, except that 0.2 g of alginic acid powder is dissolved in 50 mL of distilled water to obtain a sodium alginate solution with a concentration of 10 mmol / L. Through calculation, the alginic acid solution with the concentration of 0.125mmol / L, 0.25mmol / L, 0.5mmol / L, 1mmol / L, 2mmol / L and 3mmol / L was prepared, and L-dopa PBS buffer before adding TYR respectively Add different concentrations of alginic acid solution to the solution, keep it in a 32℃ water bath for 20 minutes, then use an ultraviolet spectrophotometer to measure the absorbance at 475nm, and explore the effect of alginic acid on the activity of TYR and the inhibition rate through the change in absorbance. . Among them, the alginic acid inhibition rate formula is:
[0052]
[0053] figure 2 It is a graph showing the inhibition of TYR activity by alginic acid.
[0054] by fi...
Embodiment 3
[0055] Example 3. Research on the mechanism of alginic acid on TYR inhibition
[0056] When alginic acid acts on TYR, the active center of TYR can emit emission light of a specific wavelength after being excited. Fluorescence spectroscopy is used to determine the effect of alginic acid inhibitors on the emission spectrum of TYR, and the effect of alginic acid inhibitors on TYR can be judged. Mechanism. The specific operation method is as follows:
[0057] Prepare PBS buffer solution with pH=6.8, add TYR solution with a concentration of 0.01mg / mL, add 1mL of alginic acid inhibitors of the same volume and different concentration, and mix them in a water bath at 32℃ for 20 minutes. Use excitation wavelength 280nm, scan each The emission spectrum of the sample between 290-450nm.
[0058] image 3 Fluorescence analysis diagram of the structure of the interaction between alginic acid and TYR.
[0059] by image 3 It can be seen that as the concentration of alginic acid continues to rise, ...
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