中国中药杂志

2021, v.46(03) 645-660

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HPLC-MS~n分析小叶莲乙酸乙酯提取物的主要成分
Identification of chemical constituents in ethyl acetate soluble extract of Sinopodophylli Fructus based on HPLC-MS~n

范珊珊;尚明英;徐风;刘广学;李耀利;蔡少青;
FAN Shan-shan;SHANG Ming-ying;XU Feng;LIU Guang-xue;LI Yao-li;CAI Shao-qing;Division of Pharmacognosy, School of Pharmaceutical Sciences, Peking University;

摘要(Abstract):

采用HPLC-MS~n联用技术对藏族药小叶莲具有抗乳腺癌活性的乙酸乙酯提取物的化学成分进行定性分析。应用Kromasil 100-5 C_(18)色谱柱(4.6 mm×250 mm,5μm),以0.1%甲酸水溶液(A)-乙腈(B)为流动相进行梯度洗脱,流速1.0 mL·min~(-1)。质谱使用ESI离子源,正、负离子分析模式采集数据,通过高分辨质谱解析,并结合相关文献和对照品数据共指认了102个化合物,包括黄酮类成分67个和木脂素类成分35个,其中小叶莲中首次报道的化合物45个(43个为黄酮类化合物,2个为鬼臼木脂素类化合物),新化合物19个(18个为异戊烯基黄酮类化合物,1个鬼臼木脂素类化合物)。利用PharmMapper对首次在小叶莲中报道的化合物进行活性预测,发现有20个化合物具有潜在的抗癌活性。该研究发现小叶莲乙酸乙酯提取物中存在较多的同分异构体,共发现同分异构体19组,其中分子式为C_(21)H_(20)O_8的化合物同分异构体最多,共有18个化合物,均为α-peltatin或其异构体;其次是C_(21)H_(20)O_7,共有10个化合物,均为8-prenylquercetin-3-methyl ether或其异构体。HPLC-MS~n技术能快速指认小叶莲具有抗乳腺癌活性的乙酸乙酯提取物中所含的化学成分,研究结果为阐释小叶莲抗乳腺癌的药效物质和作用机制提供了更多的科学依据。
A high performance liquid chromatography with a diode array detector combined with electrospray ionization ion trap time-of-flight multistage mass spectrometry(HPLC-DAD-ESI-IT-TOF-MS~n, HPLC-MS~n) method was established for qualitative analysis of the chemical components of ethyl acetate extract from Sinopodophylli Fructus. The analysis was performed on a Kromasil 100-5 C_(18)(4.6 mm×250 mm, 5 μm) column, with a mobile phase consisted of 0.1% formic acid(A) and acetonitrile(B) for gradient at a flow rate of 1.0 mL·min~(-1). Electrospray ionization ion trap time-of-flight multistage mass spectrometry was applied for qualitative analysis under positive and negative ion modes. With use of reference substance, characteristic fragmentation and their HR-MS data, 102 components were identified, including 67 flavonoids and 35 lignans. Among them, 45 compounds were reported in Sinopodophylli Fructus for the first time and 19 compounds were identified as new compounds. PharmMapper was used to predict the bioactivity of compounds that were first reported in Sinopodophylli Fructus, and 20 compounds of them were identified to have potential anticancer activity. The results showed that there were many isomers in the ethyl acetate extract of Folium Nelumbinis, and a total of 19 groups of isomers were found. Among them, C_(21)H_(20)O_8 had the highest number of isomers(18 compounds), all of which were α-peltatin or its isomers; C_(21)H_(20)O_7 ranked second, with 10 compounds, all of which were 8-prenylquercetin-3-methyl ether or its isomers. In conclusion, an HPLC-MS~n method was established for qualitative analysis of the ethyl acetate extract(with anti-breast cancer activity) from Sinopodophylli Fructus in this study, which will provide the evidence for clarifying pharmacological active ingredients of the ethyl acetate extract from Sinopodophylli Fructus against breast cancer.

关键词(KeyWords): 小叶莲;HPLC-MS~n;异戊烯基黄酮;环木脂内脂木脂素
Sinopodophylli Fructus;HPLC-MS~n;prenylflavonoid;cycloxylinolide

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(81673590)

作者(Author): 范珊珊;尚明英;徐风;刘广学;李耀利;蔡少青;
FAN Shan-shan;SHANG Ming-ying;XU Feng;LIU Guang-xue;LI Yao-li;CAI Shao-qing;Division of Pharmacognosy, School of Pharmaceutical Sciences, Peking University;

Email:

DOI: 10.19540/j.cnki.cjcmm.20200629.203

参考文献(References):

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