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 Beijing 100191, China.
Zhongguo Zhong Yao Za Zhi. 2021 Feb;46(3):645-660. doi: 10.19540/j.cnki.cjcmm.20200629.203.
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-MSn, HPLC-MSn) 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-MSn 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.
建立了一种高效液相色谱-二极管阵列检测器结合电喷雾电离离子阱飞行时间多级质谱（HPLC-DAD-ESI-IT-TOF-MSⁿ，HPLC-MSⁿ）法，用于定性分析山荷叶果实乙酸乙酯提取物的化学成分。分析在Kromasil 100-5 C₁₈（4.6 mm×250 mm，5 μm）柱上进行，流动相由0.1%甲酸（A）和乙腈（B）组成，梯度洗脱，流速为1.0 mL·min⁻¹。采用电喷雾电离离子阱飞行时间多级质谱在正离子和负离子模式下进行定性分析。通过使用对照品、特征碎片及其高分辨质谱数据，鉴定出102种成分，包括67种黄酮类化合物和35种木脂素类化合物。其中，45种化合物首次在山荷叶果实中报道，19种化合物被鉴定为新化合物。利用PharmMapper预测首次在山荷叶果实中报道的化合物的生物活性，其中20种化合物被鉴定具有潜在的抗癌活性。结果表明，山荷叶乙酸乙酯提取物中存在许多异构体，共发现19组异构体。其中，C₂₁H₂₀O₈的异构体数量最多（18种化合物），均为α-盾叶鬼臼毒素或其异构体；C₂₁H₂₀O₇排名第二，有10种化合物，均为8-异戊烯基槲皮素-3-甲醚或其异构体。总之，本研究建立了一种HPLC-MSⁿ法用于定性分析山荷叶果实具有抗乳腺癌活性的乙酸乙酯提取物，这将为阐明山荷叶果实乙酸乙酯提取物抗乳腺癌的药理活性成分提供依据。
