HPLC同时测定何首乌中5种成分及指纹图谱的建立

Simultaneous Determination of Five Components in Polygoni Multiflori Radix by HPLC and Establishment of Fingerprints

目的建立HPLC同时测定不同产地何首乌中5种指标性成分含量的方法,并建立何首乌HPLC指纹图谱分析方法。方法采用Agilent Eclipse XDB-C18色谱柱(4.6 mm×250 mm,5μm),流动相为乙腈-0.1%磷酸水溶液,梯度洗脱,测定何首乌样品中五没食子酰葡萄糖、没食子酸、二苯乙烯苷、大黄素、大黄素甲醚含量,用SPSS19.0统计软件对数据进行聚类分析,建立何首乌HPLC指纹图谱并进行分析。结果何首乌中5种指标性成分的进样量在线性范围内与峰面积积分值呈良好的线性关系,相关系数均大于0.999 4。安徽阜阳样品中五没食子酰葡萄糖、没食子酸、二苯乙烯苷、大黄素甲醚含量最高(分别为0.048 9%、0.036 1%、3.179 9%、0.551 2%),贵州安顺样品中大黄素含量最高(0.423 8%);安徽阜阳样品中指标成分平均总含量最高(3.966 6%),其次为安徽亳州(2.769 8%)、贵州(2.469 4%)及四川(2.306 3%)。云南何首乌样品与其他产地的指纹图谱差异较大,且二苯乙烯苷、大黄素、大黄素甲醚成分未检出。14批样品指纹图谱有18个共有峰,指认了2、4、7、17、18号峰分别为五没食子酰葡萄糖、没食子酸、二苯乙烯苷、大黄素、大黄素甲醚。结论本试验建立的方法稳定可靠,重现性好,能有效用于何首乌的质量控制。

Objective To establish an HPLC method to determine the contents of five kinds of index components in Polygoni Multiflori Radix from different producing areas;To establish HPLC fingerprint analysis method of Polygoni Multiflori Radix. Methods Agilent Eclipse XDB-C18 column (4.6 mm × 250 mm, 5 μm) was used, with acetonitrile - 0.1% phosphoric acid aqueous solution as the mobile phase, gradient elution, to determine the contents of galloyl glucose, gallic acid, stilbene glycoside, emodin, and emodin methyl ether in Polygoni Multiflori Radix, and SPSS19.0 software was used to conduct clustering analysis for the data. HPLC fingerprints of Polygoni Multiflori Radix were established and analyzed. Results The injection volume of five index components of Polygoni Multiflori Radix showed a good linear relationship with the peak area integral value in the linear range, and the correlation coefficient were all higher than 0.999 4. The results of the content determination showed that the contents of galloyl glucose, gallic acid, stilbene glycoside, and emodin methyl ether in Polygoni Multiflori Radix from Fuyang, Anhui Province was the highest (0.048 9%, 0.036 1%, 3.179 9%, 0.551 2%, respectively), and the content of emodin of Polygoni Multiflori Radix from Anshun, Guizhou Province was the highest (0.423 8%). The average total content of index components of sample from Fuyang, Anhui Province was the highest (3.966 6%), followed by samples from Bozhou, Anhui Province (2.769 8%), Guizhou Province (2.469 4%) and Sichuan Province (2.306 3%). The fingerprints of Polygoni Multiflori Radix samples from Yunnan Province were quite different from other producing areas, and stilbene glycoside, emodin, and emodin methyl ether were not detected. There were 18 common peaks in the fingerprints of 14 batches of samples, and the peak 2, 4, 7, 17, and 18 were determined to be galloyl glucose, gallic acid, stilbene glycosides, emodin, and emodin methyl ether. Conclusion The method established by the experiment is stable, reliable, and reproducible, and can be effectively applied to the quality control of Polygoni Multiflori Radix.