HPLC-DAD法同时测定金银花中7种多酚类物质
Simultaneous determination of seven phenolic compounds in Lonicera japonica by HPLC-DAD
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摘要: 建立了高效液相色谱-二极管阵列法(HPLC-DAD)同时测定金银花中绿原酸、咖啡酸、阿魏酸、香豆酸、肉桂酸、芦丁和木犀草苷的方法。采用岛津Shim-packVP-ODS C18(250mm×4.6mm,5μm)色谱柱,2%甲酸-2%甲酸甲醇溶液为流动相梯度洗脱,柱温35℃,流速0.7mL/min。结果表明:绿原酸、咖啡酸、阿魏酸、香豆酸、肉桂酸、芦丁和木犀草苷分别在1.04520、1.02510、1.59530、1.26630、1.89560、1.62540、0.2958mg/L范围内线性关系良好(r值范围为0.99640.9999),检出限(RSN=3)为0.0010.078mg/L,精密度RSD小于3.27%,平均回收率为93.42%102.53%。本方法操作简单,灵敏度高、重现性好,可用于金银花中7种多酚类物质的定量分析和质量监控。Abstract: A high performance liquid chromatography with diode array detection (HPLC-DAD) method was developed for the simultaneous determination of seven phenolic compounds including chlorogenic acid, caffeic acid, ferulic acid, p-coumalic acid, trans-cinnamic acid, rutin and luteolin-7-O-glucoside in Lonicera japonica.The samples were separated with a ODS-C18column (250mm×4.6mm, 5μm Shimadzu Corporation) by gradient elution using 2% formic acid in water and 2% formic acid in methanol as the mobile phase at a flow rate of0.7mL/min in 35℃. The standard curve revealed a good linear relationship (r=0.9964~0.9999) over a range of1.04~520mg/L for chlorogenic acid, 1.02~510mg/L for caffeic acid, 1.59~530mg/L for ferulic acid, 1.26~630mg/L for p-coumalic acid, 1.89~560mg/L for trans-cinnamic acid, 1.62~540mg/L for rutin and 0.29~58mg/L for luteolin-7-O-glucoside respectively. The limits of detection were ranged from 0.001 to 0.078mg/L (RSN=3) . The average recovery rates were 93.42%~102.53% and the precisions <3.27%. This method was simple, sensitive with good reproducibility and could be used for qualitative evaluation and quality control of seven phenolic compounds in Lonicera japonica.