液相色谱质谱联用测定五子衍宗方有效部位中9种成分

目的: 建立五子衍宗方乙醇提取物中9种活性成分的含量测定方法。 方法: 应用液相色谱质谱联用(LC-MS)进行测定,其中质谱采用提取离子扫描模式,其参数为:正离子,Gas temp 350 ℃,Drying Gas(N₂) 10 L·min^-1 ,Nebulizer(N₂) 1.4 MPa,Fragmentor 175 V,Skimmer 75 V,OCT 1RF Vpp 700 V,VCap 4.0 kV,质量范围 105~700 m/z。液相色谱条件为: Chromolith Fastgradient RP-18e色谱柱,50mm ×2 mm(Merck,USA) ;流动相A 相为0.1%甲酸水溶液,B 相为乙腈;流速为0.3 mL·min^-1 ;柱温为35 ℃;进样量为10 μL;运行时间为25 min。 结果: 9种成分日内和隔天精密度的RSD 均分别小于 1.5%和2.5%( n=5); 重复性RSD<2.5%( n=5);五子衍宗乙醇提取物中9种对照品的加样回收率均在96%~103%之间。 结论: 该测定方法简单、稳定,是一种较理想的含量测定方法,可用于五子衍宗方制剂的质量控制。

Objective: To establish a method for determination of 9 active compounds in alcohol extract of Wuziyanzong. Methods: The method was developed with liquid chromatogram/ tandem mass spectrometry (LC/Q-TOF),where the mass spectrometer was operated in Extractive Ion Chromatogram scan type under the positive ion mode.Nitrogen was used as drying gas,10 L·min^-1 and Nebulizer(N₂),1.4 MPa.The VCap was set at 4.0 kV,as well as the gas temperature was 350 ℃.Fragmentor and Skimmer was set at 175 V and 75 V,respectively.Chromatographic separation was achieved on a Chromolith Fastgradient RP-18e column,50 mm×2 mm( Merck,USA),using a gradient elution at a temperature of 35 ℃.The mobile phase was composed of 0.1%(v/v) acetic acid in water (A) and ACN(B) at flow-rate of 0.30 mL·min^-1. Results: Intra-and inter-day precision of 9 active compounds were under 1.5% and 2.5%(RSD,n=5),respectively;the RSD of reproducibility was less than 2.5%( n=5);the average recovery of 9 active compounds in alcohol extraction of wuziyanzong was between 96%-103%. Conclusion: The LC-MS methods are simple and reliable for the quality control of Wuziyanzong and its extracts.