基于UHPLC-MS/MS苦参实定性定量及其入血成分分析

Qualitative and quantitative determination of Sophora flavescens seeds and analysis of its blood components based on UHPLC-MS/MS

目的基于UPLC-Q-Exactive轨道阱高分辨质谱技术分析鉴定苦参实及其入血成分,采用UHPLC-MS/MS同时定量分析苦参实中6种生物碱。方法采用Waters HSS T3色谱柱(100 mm×2.1 mm,1.7μm);流动相为0.1%甲酸乙腈(10mmol/L醋酸铵)溶液(A)和0.1%甲酸水(10mmol/L醋酸铵)溶液(B),梯度洗脱;质谱采用电喷雾离子源(ESI);正离子模式下采集数据进行定性定量分析。结果苦参实样品中共鉴定34个色谱峰,大多数为生物碱类化学成分,有26个,血浆样品中共鉴定了18个生物碱以原型入血的色谱峰。苦参实中6种生物碱的总质量分数高达18.5%,其中氧化苦参碱含量最高。结论苦参实含有大量的生物碱类物质,苦参实的急性毒性可能是由于生物碱成分直接入血,本研究为苦参实的开发利用提供参考。

Objective Based on UPLC-Q-Exactive orbitrap high-resolution mass spectrometry, qualitative analysis of Sophora flavescens seeds and its blood components were analyzed, and UHPLC-MS/MS quantitative analysis of six alkaloids in Sophora flavescens seeds were performed. Methods The separation was performed on the Waters HSS T3 column(100 mm × 2.1 mm, 1.7μm), with mobile phase 0.1% formic acid acetonitrile(10 mmol/L ammonium acetate) solution(A) and 0.1% formic acid water(10 mmol/L ammonium acetate) solution(B) for gradient elution. Electrospray(ESI) fog ion source was used in mass spectrometry;Data were collected in positive ion mode for qualitative and quantitative analysis. Results A total of 34 chromatographic peaks were identified in the samples of Sophora flavescens seeds. Most of them were 26 alkaloids. A total of 18 alkaloids were identified in the plasma samples. The content of six alkaloids in Sophora flavescens seeds was up to 18.5%, and the content of oxymatrine was the highest. Conclusion The acute toxity of Sophora flavescens seeds may be due to the alkaloids entering into blood directly. This study provides a reference for the development and utilization of Sophora flavescens seeds.