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

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。

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

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 completely 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 quantitation 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 components in six different Flos Lonicerae samples with contents of 3.98-14 356.31 mg/kg.The proposed method was fast,accurate,and was suitable for the quality control of Flos Lonicerae.