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Quantitative Analysis of the Multicomponent and Spectrum–Effect Correlation of the Antispasmodic Activity of Shaoyao-Gancao Decoction

机译:韶药干草汤解痉活性的多组分及谱效相关性的定量分析

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摘要

Shaoyao-Gancao Decoction (SGD) is a well-known classic traditional Chinese medicine (TCM) with antispasmodic, anti-inflammatory, and analgesic effects. This preparation has been widely used to treat spasticity diseases in the clinic. To date, the material basis of SGD remains unclear, and the spectrum-effect correlation of its antispasmodic activity has not been reported yet. In this study, high-performance liquid chromatography (HPLC) was used to establish the fingerprint and determine the multiple components of SGD. The common peaks of fingerprints were evaluated by the similarity with the chromatographic fingerprints of the TCM. Meanwhile, the multiple components were quantified and analysed using the heatmap and box size analysis. Furthermore, data on the antispasmodic effect were extracted through in vitro smooth muscle contraction assay. Grey relational analysis combined with partial least square regression was used to study the spectrum–effect correlation of SGD. Finally, the potential antispasmolytic components were validated using an isolated tissue experiment. The HPLC fingerprint was established, and 20 common peaks were identified. The similarities of 15 batches of SGD were all above 0.965. The HPLC method for simultaneous determination of the multiple components was accurate and reliable. The contents of albiflorin, paeoniflorin, liquiritin, and glycyrrhizic acid were higher than the other components in SGD. The heatmap and box size also showed that X3 (albiflorin), X4 (paeoniflorin), X5 (liquiritin), X11 (liquirtigenin), and X16 (glycyrrhizic acid) could be used as quality indicators in the further establishment of quality standards. The spectrum–effect correlation results indicated that X4, X11, and X16 were highly correlated with antispasmolytic activity. Verification tests showed that paeoniflorin (11.7–29.25 μg/mL) and liquirtigenin (17.19–28.65 μg/mL) could significantly reduce the maximum contractile (P < 0.01). These compounds exerted concentration-dependent spasmolytic effects with the inhibitory response for acetylcholine (Ach)-evoked contraction. Thus, SGD had a significant antispasmodic effect, which resulted from the synergistic activity of its multiple components. These findings can be used for the pharmacodynamics study of SGD and are of great significance for the determination of quality markers and quality control.
机译:韶药干草汤 (SGD) 是一种著名的经典中药 (TCM),具有解痉、抗炎和镇痛作用。这种制剂已广泛用于治疗临床上的痉挛状态。迄今为止,SGD 的物质基础仍不清楚,其解痉活性的光谱效应相关性尚未报道。本研究采用高效液相色谱 (HPLC) 建立指纹图谱并确定 SGD 的多种成分。通过与中药色谱指纹图谱的相似性评价指纹图谱的共同峰。同时,使用热图和盒子尺寸分析对多个成分进行量化和分析。此外,通过体外平滑肌收缩试验提取了解痉效果的数据。采用灰色关系分析结合偏最小二乘回归研究 SGD 的谱效应相关性。最后,使用分离的组织实验验证潜在的解痉成分。建立 HPLC 指纹图谱,鉴定出 20 个常见峰。15 批 SGD 的相似度均在 0.965 以上。用于同时测定多种组分的 HPLC 方法准确可靠。白药苷、芍药苷、甘草素和甘草酸的含量高于 SGD 中的其他成分。热图和盒子大小还表明,X3 (白花素)、X4 (芍药苷)、X5 (liquiritin)、X11 (甘草素) 和 X16 (甘草酸) 可以作为进一步建立质量标准的质量指标。谱效应相关性结果表明,X4 、 X11 和 X16 与解痉活性高度相关。验证试验表明,芍药苷 (11.7–29.25 μg/mL) 和甘草素 (17.19–28.65 μg/mL) 可显著降低最大收缩力 (P < 0.01)。这些化合物发挥浓度依赖性解痉作用,对乙酰胆碱 (Ach) 诱发的收缩产生抑制反应。因此,SGD 具有显着的解痉作用,这是其多种成分协同作用的结果。这些发现可用于 SGD 的药效学研究,对质量标志物的测定和质量控制具有重要意义。

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