目的 建立同时测定金耳环药材中3,4,5-三甲氧基甲苯、3,5-二甲氧基甲苯、甲基丁香酚、马兜铃内酰胺I(AL-I)和7-甲氧基马兜铃内酰胺IV(7-OCH3-AL-IV)5种成分含量的超高速液相色谱-质谱联用(UFLC-MS/MS)方法,并研究5种成分在金耳环药材不同药用部位中的分布。方法 岛津Shimadzu Shim-pack XR-ODS Ⅲ色谱柱(2.0 mm×75 mm,1.6 μm),流动相为0.1%甲酸-甲醇,梯度洗脱,柱温40 ℃,大气压化学电离离子源(APCI),正离子检测模式,多反应监测扫描模式(MRM)测定。结果 5种成分的分离度良好,标准曲线在检测范围内均呈良好线性。金耳环药材不同部位中3,4,5-三甲氧基甲苯、3,5-二甲氧基甲苯、甲基丁香酚、AL-I和7-OCH3-AL-IV的含量范围分别是1.50~16.71、28.59~177.20、2.46~22.48、46.39~324.04、12.95~251.04 ng·mg-1;其中花和叶中7-OCH3-AL-IV的含量[(52.14~251.04)ng·mg-1]均高于根中含量[(28.54~40.73)ng·mg-1];花中AL-I的含量[(171.93~324.04)ng·mg-1]远高于根部含量[(78.44~124.56)ng·mg-1];花和叶部两种马兜铃内酰胺总含量大于根部;根茎、叶和花中甲基丁香酚的含量[(3.57~22.48)ng·mg-1]均大于同样品根中的含量[(2.46~7.29)ng·mg-1]。结论 本实验建立的方法操作简单,结果准确,具有较好的重复性和稳定性,其中金耳环中3,4,5-三甲氧基甲苯、3,5-二甲氧基甲苯和甲基丁香酚的含量为首次报道。由AL-I、7-OCH3-AL-IV和甲基丁香酚在不同部位的含量测定结果来看,金耳环应该以根入药,根茎、叶和花部位应该摘除,并谨慎使用。
Abstract
OBJECTIVE To evaluate the effectiveness and safety of Asarum insigne Diels and develop a sensitive, accurate, and efficient ultra-fast liquid chromatography coupled with triple quadrupole mass spectrometer (UFLC-MS/MS) method for simultaneous quantification of five compounds. METHODS Chromatographic separation was performed on a Shimadzu shim-pack XR-ODS Ⅲ column(2.0 mm×75 mm,1.6 μm). The mobile phase was composed of 0.1% aqueous formic acid and methanol and the flow rate was 0.2 mL·min-1. Using the established method, all components could be easily separated within 14 min. All components were detected in multiple reaction monitor mode after positive atmospheric pressure chemical ionization. RESULTS The method was validated, and had good precision, linearity, lower limit of detection, lower limit of quantification, recovery, and stability. This method was used to determine and compare the contents of five components in different medicinal parts of 11 samples, and the contents of 3,4,5-trimethoxytoluene, 3,5-dimethoxytoluene, methyleugenol in different medicinal parts of Asarum insigne were reported for the first time, which (in ng·mg-1) were as follows: 3,4,5-trimethoxytoluene: 1.5-16.71; 3,5-dimethoxytoluene: 28.59-177.20; methyleugenol: 2.46-22.48; AL-I: 46.39-324.04; 7-OCH3-AL-IV: 12.95-251.04. The content of 7-OCH3-AL-IV and total contents of two aristololactams in roots were lower than those in leaves or flower parts, and the contents of AL-I in flowers (171.9-324.0 ng·mg-1) were higher than those in roots (78.44-124.56 ng·mg-1). For methyleugenol, the contents in roots were lower than those in the other parts. CONCLUSION It is recommended to remove the leaves, flowers, and rhizomes when using Asarum insigne and use it with extreme caution. The UFLC-MS/MS method established in this study can provide guidance for the quality evaluation and safe clinical use of traditional Chinese medicine derived from Asarum insigne.
关键词
金耳环 /
超高速液相色谱-质谱联用 /
含量测定 /
3 /
4 /
5-三甲氧基甲苯 /
3 /
5-二甲氧基甲苯 /
安全性
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Key words
Asarum insigne /
UFLC-MS/MS /
content determination /
3,4,5-trimethoxytoluene /
3,5-dimethoxytoluene /
safety
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中图分类号：
R284
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参考文献
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脚注
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基金
国家自然科学基金青年科学基金项目资助(81403096)
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