[关键词]
[摘要]
目的 建立当归标准汤剂HPLC指纹图谱,为当归标准汤剂质量控制提供依据。方法 采用HPLC法建立15批当归标准汤剂指纹图谱,采用相似度评价结合聚类分析(CA)、主成分分析(PCA)、偏最小二乘判别分析(PLS-DA)等模式识别技术探讨影响当归标准汤剂质量的主要色谱峰,并同时测定这些主要色谱峰中阿魏酸、洋川芎内酯H、洋川芎内酯I、藁本内酯及色氨酸的含量。色谱条件为色谱柱Kromasil C18(250 mm×4.6 mm,5 μm),以乙腈-0.1%甲酸水溶液为流动相,梯度洗脱,体积流量1.2 mL/min,检测波长280 nm,柱温35℃。结果 建立的当归标准汤剂HPLC指纹图谱共标定出17个共有峰,15批样品相似度在0.788~0.983;CA、PCA将样品大致分为3类,结合PLS-DA,筛选出7个标志性化合物,其中指认出阿魏酸(8号峰)、藁本内酯(17号峰)、洋川芎内酯I(10号峰)、洋川芎内酯H(12号峰)。定量分析条件通过方法学验证,平均加样回收率为99.43%~104.35%。15批样品中阿魏酸为0.041%~5.596%、洋川芎内酯H为0.026%~1.583%、洋川芎内酯I为0.201%~6.461%、藁本内酯为0.126%~4.942%、色氨酸含为0.481%~2.753%。结论 建立的分析方法稳定可靠,重复性好,为当归标准汤剂及其相关制剂的质量评价提供参考。
[Key word]
[Abstract]
Objective To establish an HPLC method for the fingerprint analysis and content determination of standard decoction of Angelicae Sinensis Radix (ASR), and to provide an effective method to ensure the quality of standard decoction of ASR. Methods Fingerprint of standard decoction of ASR was established by HPLC. The similarity evaluation combined with cluster analysis (CA), principal component analysis (PCA), and partial least squares discriminant analysis (PLS-DA) were applied to explore chromatographic peak of main affecting the quality of standard decoction of ASR, and to determine the contents of ferulic acid, ligustilide H, ligustilide I, ligustilide and tryptophan. All samples were analyzed by Kromasil C18 column (250 mm×4.6 mm, 5 μm) maintained at 35℃ and eluted with acetonitrile-0.1% formic acid at the flow rate of 1.2 mL/min, and the detection wavelength was 280 nm. Results There were 17 common peaks in the HPLC fingerprint of established standard decoction of ASR, and the similarity of 15 batches of standard decoction of ASR was between 0.788 and 0.983. The samples were broadly divided into three categories by CA and PCA. Seven markers were verified by PLS-DA, and peaks 8, 17, 10, and 12 were identified as ferulic acid, ligustilide, ligustilide I, and ligustilide H, respectively. In quantitative analysis, the five components showed good regression (r2 > 0. 999 0) with linear range, the content respectively was 0.041%-5.596% in ferulic acid, 0.026%-1.583% in ligustilide H, 0.201%-6.461% in ligustilide I, 0.126%-4.942% in ligustilide, and 0.481%-2.753% in tryptophan, and the average recoveries were in the range of 99.43%-104.35%. Conclusion The analysis method established in this experiment is stable, reliable, and repeatable, which can provide reference for the quality evaluation of standard decoction of ASR.
[中图分类号]
[基金项目]
国家中医药管理局中药标准化项目(ZYBZH-C-JS-30);江苏省科技成果转化专项资金项目(BA2016104)