中国中药杂志

2018, v.43(08) 1667-1674

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基于HPLC二维指纹图谱结合化学计量分析对生地黄质量评价研究
Study on quality evaluation of Dihuang (Rehmannia glutinosa) by two-dimension HPLC fingerprints and chemometrics methods

黄春跃;欧阳丹薇;牛莉鑫;周靖;林珊;胡晓;
HUANG Chun-yue;OUYANG Dan-wei;NIU Li-xin;ZHOU Jing;LIN Shan;HU Xiao;Innovation Center of Chinese Medicine,China State Institute of Pharmaceutical Industry;

摘要(Abstract):

对22批安徽产生地黄饮片进行二维指纹图谱分析,采用Waters Atlantis~T3色谱柱(4.6 mm×250 mm,5μm),乙腈-0.1%磷酸水溶液为流动相,梯度洗脱,检测波长203 nm,柱温35℃,建立了HPLC-PDA指纹图谱;同时采用Welch Ultimate~Hilic-NH_2色谱柱(4.6 mm×250 mm,5μm),乙腈-水为流动相,梯度洗脱,建立了HPLC-ELSD指纹图谱。运用《中药色谱指纹图谱相似度评价系统》,分别确定了26个和10个色谱峰作为生地黄HPLC-PDA和HPLC-ELSD指纹图谱的共有峰。对多个来源、不同种类的41批地黄样品的二维指纹图谱进行化学计量分析,包括相似度分析、聚类分析和主成分分析,结果显示HPLC-PDA指纹图谱能很好的区分不同来源的生地黄样品,HPLC-ELSD指纹图谱能敏锐的鉴定生地黄和熟地黄。建立的生地黄二维指纹图谱方法色谱峰分离度高、化学信息丰富、指认的化学成分多,包括2个核苷类(尿苷和腺苷)、4个环烯醚萜苷类(梓醇、地黄苷D、地黄苷A及益母草苷),2个苯乙醇苷类(毛蕊花糖苷和肉苁蓉苷A)以及9个糖类成分,该方法简便易行,可用于生地黄的鉴别,也可为生地黄的质量评价提供依据。
The study is to establish the two-dimension HPLC fingerprints of Dihuang( Rehmannia glutinosa),by HPLC-PDA and HPLC-ELSD methods. The separations were performed on Waters Atlantis~ T3( 4. 6 mm × 250 mm,5 μm) and Welch Ultimate~ Hilic-NH_2( 4. 6 mm × 250 mm,5 μm) columns with the gradient elution of acetonitrile-0. 01% phosphoric acid and acetonitrile-water,respectively. The chromatographic display wavelength for PDA detector was set at 203 nm. For HPLC-ELSD,the nebulizer was set as cooling mode,the drift tube temperature was set at 60 ℃ and the gas pressure was 35. 0 psi. Based on similarity evaluation system for chromatographic fingerprint of traditional Chinese medicine,26 and 10 chromatographic peaks were determined as common components for HPLC-PDA and HPLC-ELSD fingerprints,respectively. Chemometrics analyses,such as similarity analysis; cluster analysis and principal component analysis,were performed on the common peak areas in two-dimension fingerprints for 41 batches of Dihuang from multiple sources. The results showed that the HPLC-PDA fingerprint could distinguish dried rehmannia root between different sources,and HPLC-ELSD fingerprint could differentiate dried rehmannia root from prepared rehmannia root. The two-dimension fingerprints were established with advantages of a good degree of separation,abundant chemical information and multi-components identified including two nucleosides( adenosine and uridine),four iridoid glycosides( catalpa alcohol,rehmaionoside D,rehmaionoside A and leonuride),two phenylethanoid glycosides( acteoside and cistanoside A) and nine sugars. The method is simple and practical,which could be used for the identification and quality assessment for Dihuang.

关键词(KeyWords): 生地黄;二维指纹图谱;化学计量分析;聚类分析;主成分分析
Dihuang (Rehmannia glutinosa);two-dimension fingerprints;chemometrics analysis;cluster analysis;principal component analysis

Abstract:

Keywords:

基金项目(Foundation): 国家中药标准化项目(ZYBZH-C-GZ-10);; 国家自然科学基金项目(81402824)

作者(Author): 黄春跃;欧阳丹薇;牛莉鑫;周靖;林珊;胡晓;
HUANG Chun-yue;OUYANG Dan-wei;NIU Li-xin;ZHOU Jing;LIN Shan;HU Xiao;Innovation Center of Chinese Medicine,China State Institute of Pharmaceutical Industry;

Email:

DOI: 10.19540/j.cnki.cjcmm.20180224.001

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