地黄炮制过程中HPLC-RID指纹图谱及模式识别研究

doi:10.14088/j.cnki.issn0439-8114.2022.23.017

湖北农业科学 ›› 2022, Vol. 61 ›› Issue (23): 93-96.doi: 10.14088/j.cnki.issn0439-8114.2022.23.017

地黄炮制过程中HPLC-RID指纹图谱及模式识别研究

张丽先^a, 宁二娟^a, 李智宁^a, 魏悦^a, 李飞飞^b, 张桃桃^a

河南省科学院,a.河南省纳普生物技术有限公司; b.天然产物重点实验室,郑州 450002

收稿日期:2022-01-04 出版日期:2022-12-10 发布日期:2023-01-27
通讯作者: 魏悦（1983-）,男,助理研究员,硕士,主要从事中药质量评价及创新中药的研究工作,（电话）0371-55935573（电子信箱）125186147@qq.com。
作者简介:张丽先（1989-）,女,河南洛阳人,助理研究员,硕士,主要从事中药新药研发与质量评价工作,（电话）0371-55935573（电子信箱）704271895@qq.com。
基金资助:
河南省科学院助推科技成果转化项目（210213014）; 河南省科学院科技开放合作项目（210913009）; 河南省科技攻关项目（222102310021）; 中央引导地方科技发展专项（211213004）; 河南省科学院基本科研业务费项目（220613105）

Study on HPLC-RID fingerprint chromatogram and pattern recognition in the processing of Rehmanniae radix

ZHANG Li-xian^a, NING Er-juan^a, LI Zhi-ning^a, WEI Yue^a, LI Fei-fei^b, ZHANG Tao-tao^a

a. Henan Natural Product Biotechnology Co., Ltd.; b. Key Laboratory of Natural Products, Henan Academy of Sciences, Zhengzhou 450002, China

Received:2022-01-04 Online:2022-12-10 Published:2023-01-27

摘要/Abstract

摘要： 为建立地黄（Rehmanniae radix）不同炮制品炮制过程中HPLC-RID特征图谱及多种糖含量测定方法,结合模式识别方法分析炮制过程中糖类成分变化趋势,采用Agilent ZORBAX NH₂ 色谱柱（250.0 mm×4.6 mm,5 μm）,以乙腈-纯水（70∶30）混合溶液为流动相,体积流量1.0 mL/min,柱温40 ℃,流通池温度50 ℃,建立地黄不同炮制品HPLC-RID特征图谱分析方法,外标法测定地黄不同炮制品中果糖、葡萄糖、蔗糖、棉子糖、水苏糖的含量;借助主成分分析（PCA）和聚类分析（HCA）模式识别方法分析鲜地黄-生地黄-熟地黄糖成分的差异与关联。结果表明,分析了12批地黄样品,匹配出HPLC-RID特征图谱共有峰6个,指认其中5个色谱峰,地黄片和鲜地黄无显著差异;鲜地黄中水苏糖含量最高;鲜地黄-生地黄-熟地黄炮制过程中,水苏糖含量锐减,果糖含量一直递增;PCA和HCA能较好地区分地黄不同炮制品,且结果一致。HPLC-RID特征图谱能反映地黄炮制过程中的变化,HPLC-RID法同时测定地黄中5种糖含量简便可靠,为地黄糖类成分提取工艺筛选及产品开发奠定研究基础。

关键词: 地黄（Rehmanniae radix）, 指纹图谱, 糖, 模式识别, 含量测定

Abstract: Aiming to establish the HPLC-RID characteristic chromatogram and various sugar content determination methods of different processing products of Rehmanniae radix, and analyze the change trend of sugar composition in the processing process combining pattern recognition, Agilent ZORBAX NH₂ column （250.0 mm×4.6 mm, 5 μm） was used with acetonitrile-pure water （70∶30） mixed solution as mobile phase, the flow rate was 1.0 mL/min, column temperature was 40 ℃, flow cell temperature was 50 ℃, and a HPLC-RID characteristic chromatographic analysis method for different processed products of Rehmanniae radix was established. The contents of fructose, glucose, sucrose, raffinose and stachyose in different processed products of Rehmanniae radix were determined by the external standard method. Principal component analysis （PCA） and hierarchical cluster analysis （HCA） pattern recognition methods were used to analyze the differences and associations of sugar components in different processed products of fresh Rehmanniae radix （including Rehmanniae radix slice）, Rehmanniae radix and Rehmanniae radix praeparata. The results showed that, 12 batches of Rehmanniae radix samples were analyzed, 6 common peaks in the HPLC-RID characteristic chromatogram were matched, and 5 chromatographic peaks were identified. There was no significant difference between Rehmanniae radix slice and fresh Rehmanniae radix. The content of stachyose in fresh Rehmanniae radix was the highest. In the processing from fresh to dried to prepared Rehmanniae radix, the content of stachyose decreased sharply and the content of fructose increased continuously. PCA and HCA could distinguish different processed products, and the results were consistent. It was concluded that, HPLC-RID characteristic map could reflect the changes in the processing of Rehmanniae radix. Simultaneous determination of five sugars in Rehmanniae radix by HPLC-RID method was simple and reliable, which provided research foundation for the extraction technology screening of sugar components and product development of Rehmanniae radix.

Key words: Rehmanniae radix, fingerprint chromatogram, sugars, pattern recognition, content determination

中图分类号:

R917

张丽先, 宁二娟, 李智宁, 魏悦, 李飞飞, 张桃桃. 地黄炮制过程中HPLC-RID指纹图谱及模式识别研究[J]. 湖北农业科学, 2022, 61(23): 93-96.

ZHANG Li-xian, NING Er-juan, LI Zhi-ning, WEI Yue, LI Fei-fei, ZHANG Tao-tao. Study on HPLC-RID fingerprint chromatogram and pattern recognition in the processing of Rehmanniae radix[J]. HUBEI AGRICULTURAL SCIENCES, 2022, 61(23): 93-96.

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地黄炮制过程中HPLC-RID指纹图谱及模式识别研究

Study on HPLC-RID fingerprint chromatogram and pattern recognition in the processing of Rehmanniae radix

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