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目的:从生产甘草酸的副产品甘草甜味素中寻找酪氨酸酶抑制剂。方法:采用高效液相色谱与酶抑制分析相结合,利用电喷雾MS-MS串联质谱方法及紫外光谱对活性成分进行结构确认,建立了甘草甜味素中酪氨酸酶抑制活性成分快速筛选与鉴别方法。HPLC色谱柱分离后,利用二极管阵列检测器得到各活性组分的紫外光谱图,流出液与在96孔板中与酪氨酸酶、L-多巴作用,利用酶标仪检测酶剩余活力,从而实现筛选的目的。结果:利用此方法从甘草甜味素中筛选出4个具有明显酪氨酸酶抑制活性的化合物,通过ESI-MS/MS分析和紫外光谱图,结合相关文献,实现了各化合物的快速鉴别。结论:4个化合物均为已知化合物,分别为甘草苷、18α-羟基甘草次酸甲酯、甘草查耳酮C和18α-甘草次酸。
OBJECTIVE: To search for tyrosinase inhibitors from licorice sweetener, a by-product of the production of glycyrrhizic acid. Methods: The structure of active ingredient was confirmed by electrospray ionization tandem mass spectrometry (MS-MS) coupled with ultraviolet spectrometry (HPLC-MS / MS) and enzyme inhibition analysis. A rapid screening method for the active constituents of tyrosinase in licorice sweetener Identification method. After the separation of the HPLC chromatographic column, the ultraviolet spectrum of each active component was obtained by using a diode array detector. The effluent was reacted with tyrosinase and L-dopa in a 96-well plate, and the residual activity of the enzyme was detected by a microplate reader. So as to achieve the purpose of screening. Results: Four compounds with obvious tyrosinase inhibition activity were selected from glycyrrhizin by this method. The rapid identification of each compound was achieved by ESI-MS / MS analysis and UV spectroscopy combined with related literatures. CONCLUSION: All four compounds are known compounds, including glycyrrhizin, 18α-hydroxymethyl glycyrrhetinate, licochalcone C and 18α-glycyrrhetinic acid.