[关键词]
[摘要]
目的 基于中药质量标志物(Q-marker)理念,采用UPLC-Q-TOF-MS、GC-MS以及网络药理学技术,建立与蜜麸枳壳核心功效关联的潜在Q-marker库,为探讨建立樟帮特色蜜麸枳壳饮片质量标准奠定基础。方法 采用Acquity UPLC BEH C18色谱柱(100 mm×2.1 mm,1.7 μm);以甲醇-0.1%甲酸水溶液为流动相进行梯度洗脱,电喷雾离子源(ESI),正离子模式采集数据,质谱扫描范围m/z 50~1 200,通过对照品、相对分子质量、质谱裂解规律和文献信息鉴定蜜麸枳壳醇提液化学成分;采用水蒸气蒸馏法提取挥发油与芳香水,利用气相色谱-质谱联用(GC-MS)技术对挥发油及芳香水进行检测,进样口温度260℃,传输线温度250℃,载气为氦气,体积流量1.0 mL/min,分流比10:1,进样量1 μL;程序升温设定起始柱温70℃,以2℃/min升至150℃,保持2 min,以6℃/min升至240℃,保持3 min,再以25℃/min升至300℃,保持2 min后结束,质谱图经NIST08标准质谱图库检索鉴定化学成分;利用Cytoscape 3.7.1软件构建蜜麸枳壳中化学成分与功能性消化不良(FD)治疗靶点胃泌素(GAS)、生长抑素(SS)的"饮片-成分-疾病-靶点"网络。结果 通过UPLC-Q-TOF-MS技术鉴定出55个化学成分,GC-MS技术鉴定出63个化学成分,其中的柚皮苷、新橙皮苷、圣草酚-7-O-葡萄糖苷、柚皮素、柠檬苦素、川陈皮素、橙皮素、辛弗林、诺米林、5-羟基-6,7,8,4'-四甲氧基黄酮、香风草苷、咖啡酸、(1R)-(+)-α-蒎烯、D-柠檬烯、间伞花烃、γ-松油烯16个成分可作为蜜麸枳壳饮片Q-marker库。结论 UPLC-Q-TOF-MS及GC-MS技术可快速定性分析蜜麸枳壳中化学成分,网络药理学研究可预测建立与宽中除胀功效相关的潜在Q-marker,为蜜麸枳壳饮片质量标准的建立提供依据。
[Key word]
[Abstract]
Objective Based on the idea of Q-marker, UPLC-Q-TOF-MS, GC-MS and network pharmacology techniques were used to establish a potential Q-Marker component library associated with the core efficacy of Zhangbang Aurantii Fructus with honeyed bran, in order to lay the foundation for exploring the establishment of pieces quality standards. Methods Chromatography separation was achieved on an Acquity UPLC BEH C18 column (100 mm×2.1 mm, 1.7 μm), and eluted with a gradient of methanol-water containing 0.1% formic acid. Data acquisition was carried out in electrospray ionization (ESI) under the positive ion mode, the scanning range was 50-1 200 m/z. Ingredients in alcohol extract of Aurantii Fructus with honeyed bran were identified according to reference substance, relative molecular weight, mass spectrometric cleavage rule and literature information; The volatile oil and aromatic water was extracted by steam distillation and analyzed by gas chromatography-mass spectrometry (GC-MS). The inlet temperature was 260℃, the transmission line temperature was 250℃, the carrier gas was helium, the flow rate was 1.0 mL/min, the split ratio was 10:1, the injection volume was 1 μL; The program temperature was 70℃ for the starting column, 2℃/min to 150℃ for 2 min, 6℃/min to 240℃ for 3 min, and then 25℃/min to 300℃ for 2 min. Mass spectrometry was identified by NIST08 standard mass spectrometry library to identify chemical constituents; Cytoscape 3.7.1 was used to construct a "pieces-compounds-diseases-targets" network of chemical constituents and functional dyspepsia in Aurantii Fructus with honeyed bran. Results A total of 55 chemical components were identified by UPLC-Q-TOF-MS, and 63 chemical components were identified by GC-MS. Naringin, neohesperidin, eriodictyol-7-O-glucoside, naringenin, limonin, nobiletin, hesperitin, 5-hydroxy-6,7,8,4'-tetramethoxyflavone, didymin, D-limonene, cymene, γ-terpinene nomilin, (1R)-(+)-α-pinene, caffeic acid and synephrine can be used as a Q-marker ingredient library for Aurantii Fructus with honeyed bran. Conclusion UPLC-Q-TOF-MS and GC-MS can quickly and qualitatively analyze the chemical constituents in the pieces. The network pharmacology can predict the potential Q-marker library related to efficacy of regulating vital energy and eliminating flatulence, and provide the basis for the establishment of quality standards for the pieces.
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[基金项目]
国家重点研发计划项目(2018YFC1707000);江西省自然科学基金资助项目(20192BAB215055);国家中医药管理局公益性行业科研专项(201507002-2);江西省卫生健康委员会科技计划项目(20195646);江西中医药大学中药学一流学科专项科研基金项目(JXSYLXK-ZHYAO041)