微波辅助萃取/高效液相色谱串联质谱法对山银花中活性成分含量的测定

Determination of the Active Components in Flos Lonicerae by Microwave Assistant Extraction Combined with High Performance Liquid Chromatography-Tandem Mass Spectrometry

建立了微波辅助萃取/高效液相色谱串联质谱法(MAE/HPLC-MS/MS)同时测定山银花中10种活性成分含量的方法。山银花药材采用MAE萃取,萃取溶剂为乙醇-水(7∶3),固液比1∶30,萃取温度70℃,萃取时间10 min。采用HPLC-MS/MS测定萃取液中活性成分的含量,色谱柱采用Agilent Poroshell120 SB-C18(100 mm×2.1 mm,2.7μm),以0.5%甲酸-乙腈为流动相进行梯度洗脱,负离子多重反应离子监测模式检测。在优化条件下,10种成分的定量分析在10 min内完成。结果表明,10种活性成分的线性范围为0.05～500 mg/L,相关系数(r)不低于0.996 9,检出限和定量下限分别在69～4 413μg/kg和231～14709μg/kg范围,回收率为94%～105%。采用该方法检测6个不同产地的山银花样品,10种活性成分的含量在3.98～14 356.31 mg/kg范围。该方法快速、准确,可有效地用于山银花药材的质量控制。

A method of microwave assistant extraction （MAE） combined with high performance liquid chromatography - tandem mass spectrometry （ HPLC - MS/MS） was developed for the determination of 10 active components in Flos Lonicerae. The Flos Lonicerae samples were extracted by MAE, using ethanol -H2O（7 ： 3） as solvent with a solid -liquid ratio of 1 ： 30 at a temperature of 70 ℃. The extracts were separated on an Agilent Poroshell 120 SB-C18 （100 mm × 2. 1 mm, 2.7 μm） column by gradient elution of 0.5% formic acid - acetonitrile. A triple quadrupole MS operating in multiple- reaction monitoring（MRM） mode with negative ion was used for the quantitative analysis of 10 active components. Under the optimized conditions, 10 constituents in Flos Lonicerae were analyzed com- pletely within 10 min. The linear ranges of 10 active components were in the range of 0. 05 - 500 mg/L, with correlation coefficients no less than 0. 996 9. The limits of detection and limits of quanti- ration were in the ranges of 69 -4 413 μg/kg and 231 - 14 709 μg/kg, respectively. The spiked re- coveries ranged from 94% to 105% . The method was applied in the determination of 10 active com- ponents in six different Flos Lonicerae samples with contents of 3.98 - 14 356.31 mg/kg. The pro- posed method was fast, accurate, and was suitable for the quality control of Flos Lonicerae.