目的 揭示川牛膝药材经酒炙和盐炙前后的化学成分变化规律,为修订和完善川牛膝质量标准提供参考。方法 采用Agilent Zorbax Eclipse XDB-C₁₈(250 mm×4.6 mm,5μm)色谱柱,0.1%磷酸水溶液-乙腈为流动相梯度洗脱,检测波长266 nm,体积流量0.5 mL/min,建立HPLC指纹图谱,记录各色谱峰的保留时间和峰面积数据。采用对照品指认主要色谱峰物质,采用向量夹角法计算指纹图谱相似系数,采用主成分分析和聚类分析方法进行模式识别。结果 从24批次样品中一共提取出25个主要色谱峰,指认了其中的葛根素、杯苋甾酮、大豆苷元3个色谱峰。主成分分析、聚类分析以及相似度结果显示,酒炙和盐炙对川牛膝化学成分的整体组成与其量水平具有显著影响。葛根素、杯苋甾酮、大豆苷元3个已知的药效成分在经酒炙后其量均未得到显著提升,经盐炙后仅有葛根素的量得到一定程度地提升。结论 现行药效指标成分对于川牛膝酒炙品和盐炙品的质控作用较弱,有关川牛膝炮制品的质控指标成分还需进一步筛选。实验所建立的HPLC化学指纹图谱及其分析方法可以用于川牛膝生品、酒炙品、盐炙品之间的鉴定及其质量控制。

Objective To reveal the variation regularity of chemical components in crude Cyathula officinalis and its processed products (stir-fried with yellow rice wine and salt), and promote the quality control of the herb. Methods Agilent Zorbax Eclipse XDB-C₁₈ column (250 mm×4.6 mm, 5 μm) was used with acetonitrile and 0.1% phosphoric acid solution in gradient elution mode. The detective wavelength was 266 nm and the flow rate was 0.5 mL/min. The fingerprints for 24 herbal samples were set up and 25 peaks were recorded with different retention time and peak areas. Three peaks were successfully identified as puerarin, cyasterone, and daidzein by comparing with the retention time of reference substances. The vectorial angle method was used to evaluate the similarity between fingerprints. The cluster analysis and principal component analysis were applied to studying the HPLC fingerprint and chemical pattern recognition. Results The two processing techniques, stir-frying with wine and salt, both had significant effect on herbal chemical profiles. The contents of three known active components, puerarin, cyasterone, and daidzein, were not observed except a little increasing of the content of puerarin after the crude herbs were processed with salt. Conclusion More precise active components of the processed products of C. officinalis need to future study to improve the current quality standard of C. officinalis in Chinese Pharmacopoeia. The method provided by this study is a powerful tool to identify and quality control between crude and processed C. officinalis because of its quantificational and visual evaluation system.

国家"十二五"科技支撑计划(2011BAI13B02-7)