苗药水冬瓜叶化学成分的HPLC-HESI-HRMS分析

Analysis on Chemical Components for the Leaves of the Miao Medicine Toricellia Angulata Oliv. Var.Intermedia（Harms） Hu by HPLC-HESI-HRMS

关键词:  水冬瓜叶  化学成分  HPLC-HESI-HRMS  高分辨质谱

Key words:  the leaves of Toricellia Angulata Oliv.var.intermedia(Harms) Hu  chemical components  HPLC-HESI-HRMS  HRMS

摘要:

目的 采用HPLC-HESI-HRMS技术对苗药水冬瓜叶中的化学成分进行定性分析。方法 采用Thermo Fisher Hypersil GOLD aQ C₁₈色谱柱（100 mm×2.1 mm，1.9 μm），以0.1%甲酸水溶液-乙腈（0.1%甲酸）为流动相梯度洗脱；质谱使用HESI离子源，正离子和负离子模式下采集数据。同时采用HPLC比较7-羟基香豆素、断氧化马钱苷、金丝桃苷、异槲皮苷对照品与样品中化合物17，20，24，25保留时间（t_R）差异性，比较薄层色谱荧光特征与比移值（Rf值），并参考文献中有关7-羟基香豆素、断氧化马钱苷、金丝桃苷、异槲皮苷质谱裂解碎片特征综合进行验证。结果 通过高分辨质谱正、负离子质谱信息、数据库及相关文献数据对照，共推测出苗药水冬瓜叶中38个化合物，包括3个氨基酸类化合物，5个生物碱类化合物，10个有机酸类化合物，3个苯丙素类化合物，3个芳香含氧衍生物类化合物，3个萜类化合物，6个黄酮类化合物，1个酰胺类化合物，4个其他类化合物。验证结果表明，7-羟基香豆素、断氧化马钱苷、金丝桃苷、异槲皮苷对照品分别与化合物17，20，24，25的t_R相比差异均<0.1 min；在药材样品溶液、7-羟基香豆素、断氧化马钱苷、金丝桃苷、异槲皮苷对照品色谱相应的位置上分别显相同颜色的蓝色荧光斑点，Rf值一致；文献表明化合物17，20，24，25分别与7-羟基香豆素、断氧化马钱苷、金丝桃苷、异槲皮苷具有相似的质谱裂解碎片特征。结论 初步鉴定出化合物17，20，24，25分别为7-羟基香豆素、断氧化马钱苷、金丝桃苷、异槲皮苷。HPLC-HESI-HRMS对苗药水冬瓜叶中化学成分的初步推测具有一定的科学性，为阐明苗药水冬瓜叶药材的药效物质基础提供依据。

Abstract:

OBJECTIVE To qualitatively analyze the chemical components in the leaves of the Miao Medicine Toricellia Angulata Oliv. var. intermedia(Harms) Hu by HPLC-HESI-HRMS. Methods Thermo Fisher Hypersil GOLD aQ C₁₈(100 mm×2.1 mm, 1.9 μm) column was adopted, with 0.1% formic acid water solution-methyl cyanide(0.1% formic) as the mobile phase for gradient elution; HESI ion source was used for mass spectra, the date were collected in positive and negative mode. At the same time, the difference of retention time(t_R) between 7-hydroxycoumarin, secoxyloganin, hyperin and isoquercitrin with compounds 17, 20, 24 and 25 were verified by HPLC. Compared the fluorescence characteristics of TLC with Rf values. And the characteristics of mass spectrometric fragmentation fragments of 7-hydroxycoumarin, secoxyloganin, hyperin and isoquercitrin were comprehensively verified in references. Results Thirty-eight chemical components in the leaves of the Miao Medicine Toricellia Angulata Oliv. var. intermedia(Harms) Hu were inferred by analyzing positive and negative ion spectra information of high resolution mass spectrometer with date from data base and relevant literature, including 3 amino acids, 5 alkaloids, 10 organic acids, 3 phenylpropanoids, 3 aromatic oxygen-containing derivatives, 3 terpenes, 6 flavonoids, 1 amide and 4 other compounds. The results showed that the difference of t_R of 7-hydroxycoumarin, secoxyloganin, hyperin and isoquercitrin were <0.1 min compared with compounds 17, 20,24 and 25, respectively. The same blue fluorescence spots were shown on the corresponding chromatographic positions of the medicinal sample solution, 7-hydroxycoumarin, secoxyloganin, hyperoside and isoquercitrin reference substances, and the Rf values were consistent. And the literature showed that compounds 17, 20, 24 and 25 had similar characteristics of MS fragmentation to 7-hydroxycoumarin, secoxyloganin loganin, hyperoside and isoquercitrin, respectively. Conclusion Compounds 17, 20, 24 and 25 were identified as 7-hydroxycoumarin, secoxyloganin, hyperin and isoquercitrin. It indicates that HPLC-HESI-HRMS has a scientific basis for the preliminary estimation of the chemical composition of the leaves of Toricellia Angulata Oliv. var. intermedia(Harms) Hu. It will provide basis for therapeutic material basis of the leaves of the Miao Medicine Toricellia Angulata Oliv. var. intermedia (Harms) Hu.