知母HPLC-UVD-ELSD指纹图谱的建立与多特征成分含量测定的质控方法研究
Study on Quality Control Method for Simultaneous Analysis of Fingerprint and Multi-Characteristic Components of Anemarrhenae Rhizoma<\i> by HPLC-UVD-ELSD
-
摘要: 目的 建立知母指纹图谱结合多特征成分含量测定的HPLC-UVD-ELSD分析方法，并应用于多批次市售药材的质量控制。方法 采用UltimateTMXB-C18色谱柱（4.6 mm×250 mm，4.5 μm），检测波长258 nm，ELSD漂移管温度65 ℃，载气流速1.0 L/min，建立知母指纹图谱与主含黄酮及皂苷类成分含量测定的HPLC-UVD-ELSD分析方法，运用相似度软件结合化学计量学分析对多批次药材进行质量评价并同时完成多特征成分的含量测定。结果 建立了高效、快捷的知母HPLC-UVD-ELSD指纹图谱与多特征成分含量测定的方法；18批市售药材的指纹图谱相似度在0.786~0.999，并指认了10个色谱峰。化学计量学结果表明造成18批药材差异较大的成分为新芒果苷、知母皂苷BⅡ和知母皂苷BⅢ；进而建立了同时测定新芒果苷、芒果苷、知母皂苷N、知母皂苷BⅡ和知母皂苷BⅢ 5个成分含量的方法，结果表明知母中主含知母皂苷BⅡ。结论 建立的指纹图谱结合多特征成分含量测定的HPLC-UVD-ELSD分析方法，更全面而准确地描述了知母药材化学成分整体轮廓与各类特征成分含量贡献，为提升市售知母药材的质量控制方法奠定了基础。Abstract: OBJECTIVE To establish the fingerprint and simultaneously determine the multi-characteristic components of Anemarrhenae rhizoma by HPLC-UVD-ELSD for conducting the quality analysis in the market. METHODS The chromatographic separation was conducted on an UltimateTM XB-C18 column (4.6 mm × 250 mm， 4.5 μm)， the monitoring wavelength was 258 nm， the temperature of drift tube was maintained at 65 ℃， and the carrier gas flow rate was 1.0 L/min. The fingerprint and quantitative analysis methods of flavonoids and saponins in Anemarrhenae rhizoma were set up by HPLC-UVD-ELSD. Similarity evaluation combined with stoichiometry analysis were used to evaluate the quality of multiple batches of Anemarrhenae rhizoma samples， and multi-characteristic components in Anemarrhenae rhizoma were selected for simultaneous quantification. RESULTS An efficient and convenient HPLC-UVD-ELSD analysis method was used for fingerprint and quantitative analysis of multi-characteristic components in Anemarrhenae rhizoma. Ten main characteristic peaks were chemically identified in the established fingerprint， and the similarity ranged from 0.786 to 0.999 for 18 batches of Anemarrhenae rhizoma. The results of PLS-DA indicated that neomangiferin， timosaponin BⅡ and timosaponin BⅢ were shown greatly different in 18 batches of Anemarrhenae rhizoma samples. In addition， a quantification method of neomangiferin， mangiferin， timosaponin N， timosaponin BⅡ and timosaponin BⅢ in Anemarrhenae rhizoma was accomplished. The quantitative results of 18 batches of samples showed that the content of timosaponin B Ⅲ was the highest in Anemarrhenae rhizoma. CONCLUSION The established fingerprint and quantification of multi-characteristic components based on HPLC-UVD-ELSD analysis method can more comprehensively and accurately describe the chemical profiles and contents of characteristic compounds in Anemarrhenae rhizoma， which lays a foundation for improving effective and overall quality control method of Anemarrhenae rhizoma.
-
Key words:
- Anemarrhenae rhizoma<\i> /
- flavonoids /
- saponins /
- HPLC-UVD-ELSD /
- fingerprint /
- content determination /
- quality control method
-
[1] 国家药典委员会.中华人民共和国药典：一部[S].北京：中国医药科技出版社，2015：212. [2] 龚林，邱彦，刘静，等.知母总皂苷对血管性抑郁小鼠抑郁行为和脑组织病理学的影响[J].中国药物应用与监测，2017，14（2）：88-92. [3] LU WQ，QIU Y，LI TJ，et al.Timosaponin B-Ⅱ inhibits pro-inflammatory cytokine induction by lipopolysaccharide in BV2 cells[J].Arch Pharm Res，2009，32（9）：1301-1308. [4] 丛悦，柳晓兰，余祖胤，等.知母皂苷抑制血小板聚集的活性成分筛选及构效关系分析[J].解放军医学杂志，2010，35（11）：1370-1373. [5] 吴薛明，许婷婷，潘扬，等.知母中具抑瘤活性的芒果苷的微波辅助提取工艺[J].南京中医药大学学报，2011，27（3）：260-262. [6] 钟艳梅，钟静君，石忠峰，等.知母总皂苷干预T2DM大鼠的药效学研究[J].时珍国医国药，2017，28（1）：71-74. [7] SHAN L，WU Y，YUAN L，et al. Rapid screening of chemical constituents in Rhizoma anemarrhenae<\i> by UPLC-Q-TOF/MS combined with data postprocessing techniques[J].Evid Based Complement Altern Med，2017，2017：1-14. [8] 许费昀.知母总黄酮片治疗阴虚热盛证2型糖尿病Ⅲ期临床研究[D].南京：南京中医药大学，2009. [9] 原源.知母防治缺血性脑中风的物质基础及品质评价[D].上海：第二军医大学，2005. [10] 朱建新，冯亭亭，罗飞，等.张家口地区知母药材UPLC指纹图谱研究[J].河北北方学院学报（自然科学版），2018，34（4）：11-17. [11] 张洪坤，黄玉瑶，路丽，等.基于HPLC-ELSD指纹图谱的亳州和河北知母的聚类和判别研究[J].时珍国医国药，2017，28（11）：2670-2673. [12] 魏静娜，刘征辉，赵琳琳，等.知母药材的HPLC/PDA/ELSD指纹图谱研究[J].天津中医药，2015，32（10）：618-621. [13] 李春兰，毛春芹，陆兔林，等.不同产地知母饮片中菝葜皂苷元及芒果苷的含量测定[J].南京中医药大学学报，2010，26（1）：59-61. [14] 陈素娟，张艳秋，聂静，等.不同产地知母药材的含量测定及指纹图谱研究[J].西北药学杂志，2019，34（3）：285-288. [15] CHONG J，SOUFAN O，LI C，et al. MetaboAnalyst 4.0：Towards more transparent and integrative metabolomics analysis[J].Nucleic Acids Res，2018，46：W486-W494. [16] CHONG J，WISHART DS，XIA J. Using MetaboAnalyst 4.0 for comprehensive and integrative metabolomics data analysis [J].Curr Protoc Bioinform，2019，68（1）：e86. [17] GENG JL，DAI Y，YAO ZH，et al.Metabolites profile of Xian-Ling-Gu-Bao capsule，a traditional Chinese medicine prescription，in rats by ultra performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry analysis[J].J Pharm Biomed Anal，2014，96：90-103. -
点击查看大图
计量
- 文章访问数: 439
- HTML全文浏览量: 4
- PDF下载量: 330
- 被引次数: 0
下载:
