越婢加术汤的HPLC指纹图谱建立、含量测定及化学模式识别分析

目的:建立越婢加术汤的高效液相色谱(HPLC)指纹图谱,测定其中盐酸麻黄碱和盐酸伪麻黄碱的含量,同时进行化学模式识别分析。方法:以甘草酸铵为参照,使用《中药色谱指纹图谱相似度评价系统(2012版)》建立10批越婢加术汤的HPLC指纹图谱并进行相似度评价,结合混合对照品指认共有峰;采用HPLC法测定盐酸麻黄碱和盐酸伪麻黄碱的含量;采用SPSS 26.0软件进行聚类分析,采用SIMCA 13.0软件进行主成分分析和偏最小二乘法-判别分析,并筛选影响越婢加术汤质量的差异性成分。结果:越婢加术汤指纹图谱中共有20个共有峰,与对照指纹图谱的相似度均大于0.92;共指认了2个色谱峰,分别为甘草苷(峰11)和甘草酸铵(峰18)。盐酸麻黄碱、盐酸伪麻黄碱检测质量浓度的线性范围分别为0.98~48μg/mL(r=0.9999)、1.02~51μg/mL(r=0.9992);精密度、重复性、稳定性(24 h)试验的RSD均小于2%;平均加样回收率分别为105.67%(RSD=2.88%,n=9)、104.15%(RSD=2.02%,n=9);含量分别为0.0081~0.0143、0.0025~0.0118 mg/mL。聚类分析结果显示,10批越婢加术汤可聚为3类,S1聚为一类,S3聚为一类,S2、S4~S10聚为一类。主成分分析结果显示,S3位于得分图的最右侧,S1位于得分图的右侧,S2、S4~S10位于得分图的中部。偏最小二乘法-判别分析结果与聚类分析、主成分分析基本一致,且峰9、峰3、峰12、峰8、峰19、峰18(甘草酸铵)、峰13、峰20、峰11(甘草苷)的重要性投影值均大于1。结论:所建HPLC指纹图谱和含量测定方法操作简便、准确,结合化学模式识别可用于越婢加术汤的质量控制。峰9等9个成分为影响越婢加术汤质量的差异性成分。

盐析辅助水蒸气蒸馏法对川芎挥发油提取的影响研究

目的:探讨盐析辅助水蒸气蒸馏法对川芎挥发油提取的影响。方法:采用气相色谱-质谱联用(GC-MS)仪对氯化钠、磷酸氢二钠辅助水蒸气蒸馏法和传统水蒸气蒸馏法提取的川芎挥发油成分进行分析,同时以丁烯基苯酞和藁本内酯的总含量为指标进行定量比较。结果:水蒸气蒸馏所得挥发油中得到17种化合物,挥发油提取率为0.15%,丁烯基苯酞和藁本内酯的平均总含量为4.09 mg/g;氯化钠辅助水蒸气蒸馏法得到15种化合物,挥发油取率为0.29%,丁烯基苯酞和藁本内酯的平均总含量为4.15 mg/g;磷酸氢二钠辅助水蒸气蒸馏法得到15种化合物,挥发油提取率为0.11%,丁烯基苯酞和藁本内酯的平均总含量为3.31 mg/g。结论:盐析辅助水蒸气蒸馏法与传统水蒸气蒸馏法相比,所提川芎挥发油成分差别不大,氯化钠辅助可提高出油率,但对丁烯基苯酞和藁本内酯含量无明显影响;磷酸氢二钠辅助会降低出油率,丁烯基苯酞和藁本内酯含量也稍有降低。

以蟾酥制剂为例,探讨一测多评法在中成药质量评价和控制中的应用研究

一测多评法,作为适合中药特点的质量评价模式,已被广泛用于中药材、饮片和提取物的质量评价,但其在组方复杂的成方制剂中的应用仍处于探索和研究阶段。该文基于《中国药典》2020年版收载的蟾酥质量标准中3种蟾毒配基类成分含量测定的一测多评法,选择7种代表性的蟾酥制剂,探讨了蟾酥药材中华蟾酥毒基对蟾毒灵和脂蟾毒配基的相对校正因子(0.923和1.04)直接应用于其制剂质量评价的可行性。采用与药材相同的色谱条件对制剂进行分析,结合专属性实验,5种制剂(牙痛一粒丸、喉症丸、熊胆救心丸、六神丸和牛黄消炎丸)可以直接应用相对校正因子进行含量计算,并以喉症丸为例,开展了3种成分同步测定的方法学验证,蟾毒灵、华蟾酥毒基和脂蟾毒配基的加样回收率在90.64%106.1%。采用相对校正因子0.923和1.04计算的24批蟾酥制剂中蟾毒灵和脂蟾毒配基的含量与外标法结果无显著性差异。10批六神丸样品中蟾毒灵、华蟾酥毒基和脂蟾毒配基质量分数分别为1.272.61、2.445.66、0.9883.16 mg·g^(-1);3个厂家10批喉症丸样品中蟾毒灵、华蟾酥毒基和脂蟾毒配基质量分数分别为0.7601.32、1.352.39、0.6001.55 mg·g^(-1)。该研究为蟾酥制剂质量标准的完善和提高提供数据支撑,并为拓展一测多评法在中成药质量评价和控制中的应用提供简化的研究思路和方法。

麝香保心丸中蟾酥的质量控制标准提升

目的:建立麝香保心丸中蟾毒灵、华蟾酥毒基和脂蟾毒配基含量测定的一测多评法,为完善麝香保心丸国家标准提供检测方法。方法:采用高效液相色谱法(HPLC)建立麝香保心丸中3种蟾毒配基含量的同步测定方法,并进行方法学验证,色谱条件为Nucleosil 100-5 C_(18)色谱柱(4.6 mm×250 mm,5μm),流动相乙腈-0.1%磷酸二氢钾水溶液(磷酸调节pH 3.2)(48∶52),流速0.6 mL·min^(-1),检测波长296 nm,柱温35℃;以华蟾酥毒基为内参物,建立其对蟾毒灵和脂蟾毒配基的相对校正因子,并对相对校正因子的关键影响因素进行考察,采用相对保留时间进行待测色谱峰定位,结合在线紫外光谱和超高效液相色谱-四级杆飞行时间质谱法(UPLC-QTOF/MS)提供的精确相对分子质量辅助识别待测色谱峰;采用外标法验证一测多评法测定结果的准确性。结果:建立了麝香保心丸中3种蟾毒配基的一测多评质量评价模式,华蟾酥毒基对蟾毒灵和脂蟾毒配基的相对校正因子分别为0.922和1.01;11批样品中三者总量范围33.7~36.0μg/丸,外标法和一测多评法测定结果无显著性差异。结论:建立的方法可用于麝香保心丸中蟾毒灵、华蟾酥毒基和脂蟾毒配基的质量控制,建议麝香保心丸国家标准含量测定项下蟾酥药材的质量控制新增蟾毒灵,并以这3种成分总量制定含量限度值。

Qualitation of Chuanxiong Rhizome essential oil and determination of 3-butylidenephthalide and ligustilide by GC-MS

Chuanxiong Rhizoma(rhizomes of Ligusticumchuanxiong Hort)is a classical Chinese medicine,and it is usually used for the treatment of cardiovascular diseases.The essential oil is the main active constituent of Chuanxiong.In this study,a GC-MS quantitative method based on the SIM scanning mode and batch processing analysis was established,and the total contents of 3-butylidenephthalide and ligustilide were used as the indicators to optimize three extraction methods,including hydro-distillation extraction(HDE),ultrasound-assisted extraction(UAE)and supercritical fluid extraction(SFE).Then the volatile oil obtained by three extraction methods was analyzed qualitatively.The results showed that the volatile oil obtained by the UAE had the highest contents of 3-butylidenephthalide and ligustilide,the volatile oil extracted by HDE had the most types of chemical components,and the SFE volatile oil had the highest oil yield.

Exploring the mechanism of Fu-Zi Decoction in treatment of chronic heart failure based on network pharmacology and molecular docking technology

In the present study, we aimed to explore the mechanism of Fu-Zi Decoction in the treatment of chronic heart failure(CHF) using network pharmacology. Sym Map database was used to analyze the modern medical(MM) symptoms of various medicines. The chemical components of Fu-Zi Decoction were obtained through TCMSP, ETCM database, and previous results. The targets of Fu-Zi Decoction were obtained through STITCH, Swiss Target Prediction, Target NET database, and literature. The targets for the treatment of CHF were obtained from the Dis Ge NET, GEO, and Drug Bank databases, and the common parts of the Fu-Zi Decoction targets were screened out to construct the PPI network. The PPI network was decomposed modularly, its functions were analyzed, and the KEGG pathway enrichment analysis was performed. The key target was verified by Swiss Dock for molecular docking. A total of 205 chemical components of Fu-Zi Decoction, 551 drug targets, and 521 disease targets were collected. Functional enrichment analysis revealed that it was mainly involved in biological processes, such as negative regulation of cell death, oxidative stress, and G protein-coupled receptor regulation. KEGG enrichment findings mainly involved fluid shear stress and atherosclerosis, IL-17 signaling pathway, and so on. The results of molecular docking showed that benzoylaconitine, aconitine, mesaconitine, paeoniflorin, and atractylodes Ⅲ all had a strong affinity with the core target CXCL8, suggesting that Fu-Zi Decoction could negatively regulate cell apoptosis and oxidative stress through fluid shear stress and atherosclerosis, IL-17 signaling pathway, and so on. Collectively, our data showed that Fu-Zi Decoction had a good effect on the treatment of CHF.