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基于UPLC 指纹图谱结合网络药理学及含量测定的经典名方酸枣仁汤质量评价
陈伟燕, 耿晓梅, 卜明楠, 赵韶华, 刘敏彦, 靳怡然
中国医院药学杂志 ›› 2025, Vol. 45 ›› Issue (2) : 141-147.
PDF(1919 KB)
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基于UPLC 指纹图谱结合网络药理学及含量测定的经典名方酸枣仁汤质量评价
Quality evaluation of the classical prescription Suanzaoren decoction based on ultra-high-performance liquid chromatography fingerprint， network pharmacology and quantitative analysis
({{custom_author.role_en}}), {{javascript:window.custom_author_en_index++;}}目的 基于UPLC指纹图谱、网络药理学和定量分析相结合的研究策略，准确评价酸枣仁汤的质量。 方法 建立酸枣仁汤指纹图谱，对10批酸枣仁汤进行相似度评价和共有峰鉴定；利用网络药理学方法预测酸枣仁汤药效成分，并采用分子对接进行验证；基于获得的药效物质建立酸枣仁汤的含量测定方法，用于评价酸枣仁汤的质量。 结果 通过指纹图谱分析，共得到酸枣仁汤32个共有峰，其中22种化学成分得到鉴定；通过网络药理学构建了“中药-活性成分-靶点-疾病”网络图，基于成分可测性和对照品的可获得性，选择贡献度较高的11种化合物作为药效关联物质，分子对接验证表明其与靶点具有较好的结合能力；对此11种化合物进行含量测定，方法学验证结果表明11种被测化合物在检测范围内均具有良好的线性关系（r2>0.994 0），精密度、准确度和稳定性均良好。 结论 该研究所建立的方法简单、快速、准确可靠，能够用于酸枣仁汤的药效物质筛选和质量评价，同时也为中药的活性成分筛选和质量评价研究提供了思路。
OBJECTIVE To evaluate accurately the quality of Suanzaoren Decoction （SZRD） based on the ultra-high-performance liquid chromatography （UPLC） fingerprint， network pharmacology and quantitative analysis. METHODS Firstly， the fingerprint of SZRD was established， and similarity evaluation and common peak identification were carried out through the analysis of the 10 batches of SZRD. Then， the pharmacodynamic components of SZRD were predicted by network pharmacology and verified by molecular interconnection. Finally， the content determination method of SZRD was established based on the obtained pharmacodynamic substances to evaluate the quality of SZRD. RESULTS A total of 32 common peaks were obtained through the UPLC fingerprint analysis and 22 chemical constituents were successfully identified. A network of “herbs-active components-targets-disease” was conducted through network pharmacology. Based on the measurability of components and availability of reference substances， the 11 compounds with higher contribution were finally selected as the pharmacodynamic substances for content determination. Molecular docking proved that the 11 compounds had good binding ability with the targets. The methodological validation showed that the 11 compounds exhibited a good linear relationship within the assay range （r2>0.9940）， and their precision， accuracy and stability were good. CONCLUSION The combination approach of UPLC fingerprint， network pharmacology and quantitative analysis is simple， rapid， accurate and reliable for screening pharmacodynamic substances and quality evaluation of SZRD. It also provides ideas for the screening of the active ingredients and quality evaluation of traditional Chinese medicine herbals.
酸枣仁汤 / 指纹图谱 / 网络药理学 / 分子对接 / 含量测定 / 质量评价 {{custom_keyword}} /
Suanzaoren decoction / fingerprint / network pharmacology / molecular docking / quantitative analysis / quality evaluation {{custom_keyword}} /
图2 酸枣仁汤共有峰鉴定及归属图A.UPLC-UV指纹图谱（UPLC-UV fingerprint）；B.UPLC-ELSD 指纹图谱（UPLC-ELSD fingerprint） 注（note）：P，茯苓（Poria）；GRR，甘草（Glycyrrhizae Radix et Rhizoma）；CXR，川芎（Chuanxiong Rhizoma）；AR，知母（Anmarrhema Rhizoma）；ZZS，酸枣仁（Ziziphi Spinosae Semen） Fig 2 Identification and attribution of common peaks of SZRD |
表1 化合物与靶点最低结合能 （kcal·mol–1）Tab 1 Minimum binding energy of compounds and targets （kcal·mol–1） |
| 化合物 名称 | 靶点 | 最低 结合能 | 化合物 名称 | 靶点 | 最低 结合能 |
|---|---|---|---|---|---|
| 木兰花碱 | SRC | –43.22 | 甘草苷 | ESR1 | –39.65 |
| 阿魏酸 | ACE | –31.32 | 洋川芎内酯A | IL-1β | –32.60 |
| 甘草素 | SRC | –38.51 | 洋川芎内酯Ⅰ | NR3C1 | –46.31 |
| 藁本内酯 | PTGS2 | –34.85 | 斯皮诺素 | TP53 | –58.12 |
| 绿原酸 | CASP3 | –41.61 | 知母皂苷BⅡ | STAT3 | –8.90 |
| 隐绿原酸 | APP | –48.37 |
表2 待测化合物的线性关系、检测限和定量限结果Tab 2 The linearity， limit of detection and limit of quantitation of the analytes |
| 化合物名称 | 线性关系 | r2 | 范围/ng | 检测限/（μg·mL–1） | 定量限/（μg·mL–1） |
|---|---|---|---|---|---|
| 绿原酸 | y=11 945x ‒ 2 351.7 | 0.999 8 | 4.92～24.58 | 0.049 2 | 0.245 8 |
| 隐绿原酸 | y=8 036x ‒ 942.4 | 0.999 6 | 1.47～7.36 | 0.073 5 | 0.294 0 |
| 木兰花碱 | y=4 441x ‒ 817.1 | 0.999 3 | 5.44～27.18 | 0.108 7 | 0.271 8 |
| 阿魏酸 | y=24 011x ‒ 1 701.7 | 1.000 0 | 2.66～13.32 | 0.026 7 | 0.133 3 |
| 斯皮诺素 | y=6 506x ‒ 1 238.5 | 1.000 0 | 7.99～39.95 | 0.079 9 | 0.266 3 |
| 甘草苷 | y=6 966x ‒ 1 068.9 | 1.000 0 | 4.79～23.95 | 0.095 8 | 0.479 0 |
| 洋川芎内酯Ⅰ | y=21 163x ‒ 666.2 | 1.000 0 | 3.88～19.38 | 0.038 8 | 0.129 2 |
| 洋川芎内酯A | y=214.36x ‒ 468.87 | 0.999 7 | 42.5～255.0 | 0.283 3 | 0.850 0 |
| 甘草素 | y=9 868.5x ‒ 2 433.1 | 1.000 0 | 4.80～24.00 | 0.048 0 | 0.240 0 |
| 藁本内酯 | y=312.81x+114.4 | 0.998 5 | 5.90～17.70 | 0.062 3 | 0.248 9 |
| 知母皂苷BⅡ | y=1.508 6x+6.447 4 | 0.994 1 | 128.2～641.0 | 24.995 0 | 49.990 0 |
表3 待测化合物方法学考察结果Tab 3 Results for methodological investigation of the analytes |
| 化合物名称 | 精密度 | 重复性 | 稳定性 | 加样回收率 | |
|---|---|---|---|---|---|
| RSD/% | RSD/% | RSD/% | 平均值/% | RSD/% | |
| 绿原酸 | 1.30 | 2.06 | 0.80 | 98.46 | 1.77 |
| 隐绿原酸 | 1.13 | 1.28 | 1.12 | 98.77 | 2.07 |
| 木兰花碱 | 0.24 | 0.78 | 0.43 | 98.10 | 2.09 |
| 阿魏酸 | 0.15 | 0.76 | 0.21 | 99.20 | 2.52 |
| 斯皮诺素 | 0.14 | 0.85 | 0.19 | 98.85 | 1.05 |
| 甘草苷 | 0.12 | 0.81 | 0.91 | 98.46 | 1.96 |
| 洋川芎内酯Ⅰ | 0.42 | 0.73 | 0.20 | 98.24 | 1.22 |
| 洋川芎内酯A | 1.09 | 2.05 | 1.35 | 100.88 | 1.46 |
| 甘草素 | 0.07 | 0.85 | 0.27 | 99.05 | 1.77 |
| 藁本内酯 | 1.05 | 1.96 | 0.31 | 98.91 | 2.14 |
| 知母皂苷BⅡ | 1.86 | 2.86 | 2.24 | 97.01 | 1.66 |
表4 10批样品中11种待测物的含量（μg·g–1）Tab 4 Contents of the 11 constituents in the 10 batches of samples （μg·g–1） |
| 化合物 | S1 | S2 | S3 | S4 | S5 | S6 | S7 | S8 | S9 | S10 | RSD/% |
|---|---|---|---|---|---|---|---|---|---|---|---|
| 绿原酸 | 202.51 | 192.57 | 170.38 | 130.15 | 231.37 | 190.81 | 191.71 | 164.42 | 210.14 | 106.10 | 21.02 |
| 隐绿原酸 | 81.13 | 75.02 | 94.55 | 96.95 | 98.21 | 94.89 | 80.66 | 67.84 | 81.64 | 59.50 | 15.84 |
| 木兰花碱 | 477.76 | 459.59 | 362.99 | 442.98 | 419.32 | 374.95 | 457.86 | 461.25 | 470.69 | 377.64 | 10.13 |
| 阿魏酸 | 150.56 | 179.76 | 147.60 | 154.09 | 184.15 | 205.08 | 169.87 | 163.99 | 160.90 | 143.08 | 11.60 |
| 斯皮诺素 | 709.81 | 703.55 | 639.10 | 668.94 | 706.13 | 593.39 | 740.38 | 686.20 | 700.81 | 621.11 | 6.77 |
| 甘草苷 | 415.36 | 622.37 | 512.93 | 492.65 | 548.75 | 579.11 | 563.20 | 541.30 | 492.00 | 556.91 | 10.75 |
| 洋川芎内酯Ⅰ | 116.67 | 163.40 | 119.23 | 145.63 | 168.07 | 140.48 | 170.88 | 152.07 | 133.38 | 162.74 | 13.35 |
| 洋川芎内酯A | 3 342.4 | 1 725.1 | 2 107.2 | 2 850.3 | 2 405.0 | 3 340.4 | 1 724.6 | 2 828.9 | 3 043.5 | 2 233.6 | 23.80 |
| 甘草素 | 118.81 | 120.56 | 110.90 | 99.05 | 102.01 | 104.44 | 107.47 | 105.65 | 118.47 | 101.06 | 7.30 |
| 藁本内酯 | 171.79 | 116.71 | 150.25 | 124.83 | 149.18 | 118.80 | 199.27 | 113.71 | 148.82 | 113.63 | 20.37 |
| 知母皂苷BⅡ | 12 949 | 12 794 | 13 474 | 12 011 | 15561 | 10 496 | 15 633 | 11 347 | 12 766 | 11 342 | 13.31 |
| 总计 | 18 736 | 17 153 | 17 889 | 17 217 | 20573 | 16 238 | 20 039 | 16 632 | 18 326 | 15 817 | 8.82 |
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A sensitive, fast and comprehensive method for the quality assessment of (SZS) standard decoction with characterization of its chemical components was developed and validated. UPLC-Q/TOF-MS/MS system was used to identify thirty-six chemical components of SZS standard decoction which included nucleosides, phenolic acids, alkaloids, and flavonoids. Furthermore, a UPLC-PDA method was validated to simultaneously determine adenosine, protocatechuic acid, magnoflorine, catechin, protocatechin, vicenin II, spinosin, kaempferol-3-rutinoside, and 6'''-feruloylspinosin which represent four species of characteristic compounds. The qualitative method had been validated according to Chinese Pharmacopoeia (2015 edition) in terms of lineary, repeatability, recovery and stability for all analytes, with the results showing good precision, accuracy and stability. In conclusion, the method using UPLC combined with MS and PDA provided a novel way for the standardization and identification of SZS standard decoction, and also offered a basis for qualitative analysis and quality assessment of the preparations for SZS standard decoction.© 2019 Xi'an Jiaotong University. Production and hosting by Elsevier B.V.
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PDF(1919 KB)
图1 10批酸枣仁汤指纹图谱图2 酸枣仁汤共有峰鉴定及归属图图3 酸枣仁汤中“中药-活性成分-靶点-疾病”网络图
表1 化合物与靶点最低结合能 （kcal·mol–1）表2 待测化合物的线性关系、检测限和定量限结果表3 待测化合物方法学考察结果表4 10批样品中11种待测物的含量（μg·g–1）
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