超高效液相色谱-串联质谱法同时分析铁皮石斛茎部鲜样中的酚酸类物质的技术及应用

Simultaneous Analysis of Phenolic Acids in Fresh Stem Samples of Dendrobium Officinale by Ultra-high Performance Liquid Chromatography-tandem Mass Spectrometry and Its Application

本文建立了超高效液相色谱-串联质谱法同时、快速检测铁皮石斛茎部鲜样中13种酚酸类化合物的方法。样品采用甲醇水溶液提取后,以0.1%甲酸-5 mmol/L乙酸铵水溶液和纯乙腈为流动相体系,经Waters T3 C18柱(50 mm×2.1 mm,1.8μm)梯度洗脱,使用Waters三重四极杆质谱仪负离子扫描多反应监测模式测定目标化合物。结果表明,13种酚酸类化合物在相应的质量浓度范围内线性关系良好,相关系数均大于0.996;方法检出限为0.5~5.6μg/L,定量限为1.5~15.2μg/L;加标实验回收率为74.8%~95.2%,相对标准偏差为1.0%~6.9%。运用所建立的方法在铁皮石斛茎部鲜样中共检出7种酚酸,其含量在0.187~38.523 mg/kg之间;其中阿魏酸含量最高,其次为芥子酸、丁香酸、水杨酸等,对香豆酸的含量最低。该方法快速、准确,可用于铁皮石斛中多种酚酸类组分的同时测定。

In this paper,an ultra-high performance liquid chromatography-tandem mass spectrometry method was developed for the simultaneous and rapid detection of 13 phenolic acid compounds in fresh samples of Dendrobium officinale stems.After the samples were extracted by picking methanol-water solution,the samples were extracted with 0.1%(V:V)formic acid-5 mmol/L ammonium acetate aqueous solution and pure acetonitrile as the mobile phase system,then the samples were eluted by a Waters T3 C18 column(50 mm×2.1 mm,1.8μm)gradient.The target compounds were determined using a Waters triple quadrupole mass spectrometer in negative ion scanning multiple reaction monitoring mode.The results suggest that the linearity of the 13 phenolic acids was good in the corresponding mass concentration range with the correlation coefficients greater than 0.996.The limits of detection of the method were 0.5~5.6μg/L and the limits of quantification were 1.5~15.2μg/L.The spike recoveries of the compounds were 74.8%-95.2%with the relative standard deviations of 1.0%-6.9%.A total of seven phenolic acids were detected in the fresh samples of Dendrobium officinale stems using the established method,and their contents ranged from 0.187 and 38.523 mg/kg.Among them,the content of ferulic acid was the highest,followed by sinapic acid,syringic acid and salicylic acid,the content of coumaric acid is the lowest.The method is fast,accurate and can be used for the simultaneous determination of multiple phenolic acid components in Dendrobium officinale.