LC-MS/MS法测定苦参提取物中苦参碱和氧化苦参碱的质量浓度

LC-MS/MS Determination of the Concentration for Matrine and Oxymatrine in Sophora Flavescens Extract

目的:探讨液相色谱/串联质谱法(Liquid chromatography-tandem mass spectrometry,LC-MS/MS)测定苦参提取物中苦参碱和氧化苦参碱的质量浓度。方法:验证LC-MS/MS测定苦参提取物中苦参碱、氧化苦参碱的可行性与有效性,并判定检出限、重复性、稳定性、加样回收率。结果绘制苦参碱、氧化苦参碱的检出标准曲线。结果:LC-MS/MS检测苦参提取物中苦参碱与氧化苦参碱的一级质谱图和二级质谱图显示,苦参碱和氧化苦参碱的相对分子量为:248.36、246.36。色谱图显示苦参碱与氧化苦参碱无干扰色谱峰,各成分及内标峰形良好,表明该方法专属性强。在5μg/L500μg/L浓度范围内,苦参碱、氧化苦参碱均呈现良好的线性关系,线性方程分别为y=6.10x-118.29和y=18.29x-14.22,检出限均为0.1μg/L。LC-MS/MS法测定苦参提取物中苦参碱、氧化苦参碱的重复性、稳定性、加样回收率RSD均<5%。结论:LC-MS/MS法测定苦参提取物中苦参碱和氧化苦参碱的质量浓度具有可行性、高灵敏度、高特异性等优点,可用于苦参提取物的质量控制与评价,并为后续他人的研究提供关键试验参考条件。

Objective: To investigate the liquid chromatography/tandem mass spectrometry(LC-MS/MS) method for the determination of the concentration of Matrine and Oxymatrine in Sophora flavescens extract. Methods: To verify the feasibility and effectiveness of the determination of matrine and oxymatrine in the extract of Sophora flavescens by LC-MS/MS, and to determine the detection limit, repeatability, stability, and recovery rate of the sample addition method, and draw the bitterness Standard curve for detection of matrine and oxymatrine. Results: LC-MS/MS detection of matrine and oxymatrine in Sophora flavescens extract showed that the relative molecular weights of matrine and oxymatrine were 248.36 and 246.36. The chromatogram shows that matrine and oxymatrine have no interference chromatographic peaks, and the peak shapes of the components and internal standard are good, indicating that the method is highly specific. In the concentration range of 5 μg/L~500 μg/L, both matrine and oxymatrine showed a good linear relationship, the linear equations were y=6.10 x-118.29 and y=18.29 x-14.22, and the detection limit were all 0.1 μg/L. The RSD LC-MS/MS method to determine the reproducibility, stability and recovery rate of matrine and oxymatrine in the extract of Sophora flavescens were all less than 5 %. Conclusion: LC-MS/MS method has the advantages of feasibility, high sensitivity and high specificity for determining the concentration of matrine and oxymatrine in Sophora flavescens extract. It can be used for the quality control and evaluation of Sophora flavescens extract, and subsequent studies by others provide key experimental reference conditions.