漆树木材中漆黄素的分离及其抑制酪氨酸酶活性的研究

Separation of Fisetin from Toxicodendron vernicifluum( Stokes)F. A. Barkl. Woods and Its Anti-tyrosinase Activity

采用体积分数70%乙醇水溶液对漆树木材进行减压回流提取,经过乙酸乙酯萃取和中低压色谱快速分离得到纯度70%以上的化合物,并进一步采用高效液相色谱纯化,分离得到单体化合物。经过IR、MS和NMR鉴定,化合物为漆黄素。进一步研究了漆黄素对酪氨酸酶活性的抑制作用。结果表明,漆黄素对多巴色素的形成有较强的抑制作用,对酪氨酸酶单酚酶的半数抑制浓度(IC50)为69.46 mg/L。对酪氨酸酶单酚酶的抑制表现出迟滞效应,0.524 mmol/L的漆黄素能将迟滞效应由1 min延长至11.2 min。通过Lineweaver-Burk图显示漆黄素对二酚酶的抑制作用表现为竞争性抑制。

The Toxicodendron vernicifluum （Stokes） F. A. Barkl. woods were extracted with 70 % ehanol by decompressing inner ebullition method. The compound with the purity over 70 % was obtained through the extraction by rapid isolation of extracts with ethyl acetate and middle pressure chromatography. Then the compound was separated by HPLC to obtain monomeric compound. The compound was identified as fisetin by IR, MS and NMR. The inhibitories of fisetin on the activities of monophenolase and diphenolase contained in tyrosinase were studied. The results showed that fisetin could potently inhibit the generation of dopachrome. The half inhibitory concentration of monophenolase was 69.46 mg/L. The inhibitory of fisetin against monophenolase exhibited retardation effect. Fisetin with 0. 524 mmoL/L could extend the lag time from 1 min to 11.2 min. Fisetin functioned as a competitive inhibitor against diphenolase by Lineweaver-Burk plots.