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芸香科柑橘属立花橘(Citrus tachibana)果皮的乙醇提取物对人黑素瘤细胞生成黑色素有抑制作用,本次对其活性成分进行了分离。 方法:①抑制黑色素生成活性:将人黑素瘤细胞(HM3KO)接种于平皿,细胞粘附后加受试物培养5d,其后用胰蛋白酶处理细胞,离心制备细胞颗粒观察色调的变化。进一步将细胞溶解于碱性溶液,在410nm检测吸光度。②活性成分的分离:橘皮的乙醇提取物用乙酸乙酯-水分配,具有活性成分的乙酸乙酯部分,在确认其活性的同时,用硅胶柱色谱、ODS制备型HPLC分离活性成分。
The ethanol extract of the Citrus tachibana peel of Rutaceae has an inhibitory effect on the melanin production of human melanoma cells. This time, the active ingredients were isolated. Methods: 1Melanin production activity was inhibited: human melanoma cells (HM3KO) were seeded on petri dishes, cells were adhered and tested for 5 days. After that, cells were treated with trypsin and pellets were prepared by centrifugation to observe changes in hue. The cells were further dissolved in an alkaline solution and the absorbance was detected at 410 nm. 2 Isolation of active ingredient: The ethanol extract of cellulite was partitioned with ethyl acetate-water, and the ethyl acetate portion of the active ingredient was allowed to separate. The activity of the active ingredient was confirmed by silica gel column chromatography and ODS preparative HPLC.