UPLC-MS/MS法同时测定何首乌中二苯乙烯、蒽醌、黄酮及酚酸类成分

Simultaneous Determination of Stilbenes,Anthraquinones,Flavonoids and Phenolic Acids in Polygoni Multiflori Radix by UPLC-MS/MS

建立了超高效液相色谱-串联质谱（UPLC-MS/MS）同时测定何首乌中二苯乙烯（二苯乙烯苷、虎杖苷、白藜芦醇）、蒽醌（大黄素、大黄素甲醚、大黄素-8-β-D-吡喃葡萄糖苷、芦荟大黄素、大黄酸）、黄酮（表儿茶素、芦丁、金丝桃苷、紫云英苷、槲皮素）和酚酸（没食子酸）等4类共14种目标成分的方法。何首乌样品用70%甲醇在室温下以料液比1∶25超声提取45min,提取液以12 000r/min离心10min,取上清液,经0.22μm微孔滤膜过滤。采用Waters BEH C18色谱柱（100mm×2.1mm×1.7μm）,以水（含0.1%甲酸）-乙腈为流动相,梯度洗脱,柱温35℃,体积流量0.25mL/min,电喷雾离子源（ESI）,多反应离子监测（MRM）扫描方式进行检测。结果表明：14种目标化合物在一定浓度范围内均呈现良好的线性关系,线性相关系数大于0.991 5;精密度、重复性和稳定性良好;加样回收率在95.85%～102.13%之间,RSD均小于5%。该方法操作简便、灵敏度高、重现性好,可用于何首乌中多元功效物质或指标成分的同时测定,为何首乌药材内在质量的综合评价和全面控制提供新的方法参考。

A comprehensive analytical method based on UPLC-MS/MS was developed for the simultaneous determination of fourteen components include three stilbenes（stil-beneglucoside,polydatin,resveratrol）,five anthraquinones（emodin,physcion,emodin-8-β-D-glucopyranoside,aloe-emodin,rhein）,five flavonoids（epicatechin,quercetin,rutin,astragalin,hyperoside）,and one phenolic acid（gallic acid）in Polygoni Multiflori Radix.The samples was extracted with 70% methanol water solutions and assisted by ultrasonic machine.The extraction conditions which were optimized by experiment as following：the solid liquid ratio was 1∶25,the extraction time was 45 min at room temperature,the extractant phase was separated by centrifugation at 12 000r/min for 10 min,then filtered through 0.22μm nylon membranes.Under the optimized chromatographic conditions,good separation for fourteen target compounds were obtained on a Waters BEH C18（100mm×2.1mm×1.7μm）eluted by a mobile phase of water（0.1% acetic acid,A）-acetonitrile（B）at a flow rate of 0.25mL/min.The target compounds were analyzed by multiple reaction monitoring（MRM）mode.The results show that fourteen components have good linearity,the linearity of the detected components is excellent more than 0.991 5,and the limits of detection are all satisfied.The average recoveries of standard addition for the compounds are between 95.85% and 102.13%,and the relative standard deviations are less than 5%.The established method is simplify,sensitive and accurate,which is suitable for determination of fourteen components in Polygoni Multiflori Radix,and also can provide a reliable and effective technique for the quality control of Polygoni Multiflori Radix.