经典名方当归补血汤物质基准质量评价研究
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1.云南中医药大学中药学院;2.云南中医药大学

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云南省生物医药重大专项 (202202AA100009, 202302AA310006), 云南高校工程研究中心 (2020YGG01), 云南省教育厅重点实验室 (2022YGZ02)


Study on quality evaluation of classical prescription substance benchmarks of Danggui Buxue Decoction
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1.Department of Chinese Medicine,Yunnan University of Chinese Medicine;2.Yunnan University of Chinese Medicine

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    摘要:

    目的 建立经典名方当归补血汤物质基准HPLC特征图谱及多指标成分的含量测定方法,为其物质基准的质量评价及后续S开发研究提供参考。方法 采用Waters Symmetry C18色谱柱(250mm×4.6mm,5 μm),乙腈-0.1 %甲酸水溶液为流动相进行梯度洗脱,建立15批当归补血汤物质基准的HPLC特征图谱,进行相似度分析,并对共有峰进行归属,运用聚类分析(CA)、主成分分析(PCA)和偏最小二乘判别分析(PLS-DA)进行质量分析,并对其中5个指标成分进行含量测定。结果 15批当归补血汤物质基准HPLC特征图谱相似度均>0.93,确定共有峰8个。聚类分析、主成分分析可将样品分为3类,偏最小二乘判别分析筛选出4个质量差异标志物。5个成分毛蕊异黄酮葡萄糖苷、阿魏酸、芒柄花苷、毛蕊异黄酮、芒柄花素在其各自进样浓度范围内与峰面积呈良好线性关系(r>0.999),平均加样回收率为98.62 %~103.49 %,RSD均<2.00 %,含量分别为0.357~1.286、0.072~1.138、0.115~0.684、0.068~0.544、0.026~0.297 mg/g。结论 本研究建立的方法操作简单、专属性强、分离度好、灵敏度高,可用于经典名方当归补血汤物质基准质量控制和评价。

    Abstract:

    ABSTRACT:Objective To establish HPLC characteristic chromatogram and content determination methods for multi index components of Danggui Buxue decoction substance benchmark, providing reference for quality control and subsequent development and research of this famous classical prescriptions' substance benchmark. Methods The separation was performed on Waters Symmetry C18 column (250 mm × 4.6 mm, 5 μm) for gradient elution with acetonitrile-0.1% formic acid aqueous solution, and detection wavelength 260 nm. Then, HPLC fingerprint spectra of 15 batches of Danggui Buxue decoction substance standards were established, after that, analyzed similarities and assigned common peaks. Using cluster analysis (CA), principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA) for quality analysis, and conducted quantitative analysis on 5 of the indicator components. Results The HPLC fingerprint of 15 batches of Danggui Buxue decoction identified 8 common peaks with a similarity of > 0.93. Cluster analysis and principal component analysis could divide the samples into three categories, and partial least squares discriminant analysis might screen out four quality difference biomarkers. The linear relationship between the five components of calycosin-7-O-β-D-glucosid, ferulic acid, ononin, calycosin, and formononetin was good within their respective injection volume ranges, with correlation coefficients>0.999. The average sample recovery rates were 98.62% - 103.49%, and the RSD was<2.00%, The contents of 5 components ranged from 0.357-1.286, 0.072-1.138, 0.115-0.684, 0.068-0.544, 0.026-0.297 mg/g, respectively. Conclusion The method established in this study has simple operation, strong specificity, good separation, and high sensitivity, can be used for the reference quality control of Danggui Buxue decoction substance benchmark.

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  • 收稿日期:2024-02-20
  • 最后修改日期:2024-03-12
  • 录用日期:2024-03-14
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