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

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。 

Study on Quality Control Method for Simultaneous Analysis of Fingerprint and Multi-Characteristic Components of Anemarrhenae Rhizoma<\i> by HPLC-UVD-ELSD

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.