Caffeoyl hydroquinone ester and preparation method and application of caffeoyl hydroquinone ester in preparation of tyrosinase inhibitor
A technology of tyrosinase and inhibitors, applied in the field of a new natural compound of caffeoyl p-hydroxyphenol ester, which can solve pigmentation, dark spots, age spots, reduce storage period, nutritional value and market value, pigmentation Diseases and other problems, to achieve the effect of wide application potential and rich material sources
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[0028] Example 1: Preparation of caffeoyl p-hydroxyphenyl caffeate from Wedelia chinensis
[0029] 1.1 Instruments and reagents
[0030] Decompression concentration adopts N-1000 rotary evaporator, CCA-1110 circulating cooling box and SB-1000 electric thermostatic water bath from Tokyo Physicochemical Company; MPLC adopts CXTH P3000 liquid chromatograph of Chuangxin Tongheng Company, UV 3000UV-Vis detection And C18 reverse phase chromatography column (particle size 50μm, YMC Co. Ltd., Kyoto, Japan, 400mm×25mm); electrospray mass spectrometry (ESIMS) adopts MDS SCIEX API 2000 LC / MS / MS instrument from Applied Biosystems Methanol is used as the solvent for direct sample injection determination; 1 H NMR spectrum and 13 The C NMR spectrum was measured by Bruker AVANCE 600 nuclear magnetic resonance instrument with tetramethylsilane as the internal standard. The color development method adopts 10% sulfuric acid ethanol solution to heat and develop color.
[0031] 1.2 Plant source and ide...
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[0042] Example 2: Detection of tyrosinase inhibitory activity of compound p-hydroxyphenyl caffeate
[0043] 2.1 Reagents and instruments
[0044] Reagents: Tyrosinase was purchased from Sigma Chemical Co. (Sigma-Aldrich, St. Louis, USA), levodopa (L-DOPA), Kojic acid were purchased from Aladdin Industrial Corporation (USA), Na 2 HPO 4 , NaH 2 PO 4 , The compound to be tested is prepared by phytochemical analysis experiment;
[0045] Experimental instrument: Microplate reader (Genois microplate reader, Tecan GENios, Swizerland).
[0046] 2.2 Test method
[0047] a) Preparation of drug solution: The test compounds p-hydroxyphenyl caffeate and kojic acid are respectively prepared from dimethyl sulfoxide (DMSO) into a solution of 10mg / ml, 67mmol of phosphate buffer (prepared with ultrapure water), 46U / ml Tyrosinase solution (prepared with phosphate buffer), 2.5mM L-DOPA (prepared with phosphate buffer).
[0048] b) Using a colorimetric method, complete the determination of the half inhibito...
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