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

摘要

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 Poroshell 120 SB-C18( 100 mm × 2.1 mm,2.7 μm),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0.9969,检出限和定量下限分别在69～4413 μg/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-tation were in the ranges of 69 -4 413 μg/kg and 231 - 14 709 μg/kg, respectively. The spiked recoveries 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.