豨莶草HPLC-CAD指纹图谱的建立及多成分含量测定

Establishment of an HPLC-CAD fingerprint and multi-component content determination of Siegesbeckiae Herba

关键词:  豨莶草  高效液相色谱  电喷雾检测器  指纹图谱  含量测定  奇壬醇

Key words:  Siegesbeckiae Herba  high performance liquid chromatography  charged aerosol detector  fingerprint  content determination  kirenol

摘要:

[目的] 建立豨莶草药材高效液相色谱串联电喷雾检测器（HPLC-CAD）指纹图谱及多成分同时定量的整合分析方法，为豨莶草药材的质量评价提供参考。[方法] 采用Eclipse Plus C₁₈色谱柱（4.6 mm×250 mm，5 μm），以0.1%（v/v）甲酸水（A）-乙腈（B）为流动相进行梯度洗脱，进样量10 μL，柱温35 ℃。CAD雾化器温度35 ℃，采集频率10 Hz，过滤常数5.0 s。建立不同产地的21批豨莶草HPLC-CAD指纹图谱，并对豨莶草药材中8个指标性成分的含量进行测定。[结果] 21批豨莶草指纹图谱相似度为0.912 ~ 0.978，标定了17个共有峰，并指认出8个共有峰，分别为绿原酸、芦丁、3，4-二咖啡酰奎宁酸、4，5-二-O-咖啡酰奎宁酸、奇壬醇、豨莶苷、Hythiemoside B、豨莶精醇。8个成分在各自浓度范围内线性关系良好（r≥0.999 3），方法精密度、重复性，样品稳定性良好，相对标准偏差（RSD）均小于5.00%，平均加样回收率为100.87%~102.99%，RSD≤1.71%。[结论] 研究建立的HPLC-CAD指纹图谱和多成分含量测定方法准确、稳定，可有效评价豨莶草质量。

Abstract:

[Objective] To establish an integrated analytical method combining HPLC-CAD fingerprint and simultaneous multi-component quantification of Siegesbeckiae Herba，and to provide a reference for its quality evaluation. [Methods] An Eclipse Plus C18 column（4.6 mm×250 mm，5 μm） was used with a mobile phase consisting of 0.1%（v/v） formic acid in water（A） and acetonitrile（B） for gradient elution. The injection volume was 10 μL，and the column temperature was 35 ℃. The CAD atomizing chamber temperature was 35 ℃，the acquisition frequency was 10 Hz，and the filter constant was 5.0 s. HPLC-CAD fingerprints of 21 batches of Siegesbeckiae Herba from different regions were established，and the contents of eight indicator components were determined. [Results] The similarities of the 21 batches of fingerprints ranged from 0.912 to 0.978，and 17 common peaks were assigned. Among them，eight common peaks were identified as chlorogenic acid，rutin，3，4-dicaffeoylquinic acid，4，5-di-O-caffeoylquinic acid，kirenol，darutoside，hythiemoside B，and darutigenol. The eight components showed good linearity within their respective concentration ranges（r≥00.999 3）. The method exhibited good precision，repeatability，and sample stability，with relative standard deviations（RSD） all below 5.00%. The average recovery rates ranged from 100.87% to 102.99%，with RSDs≤1.71%. [Conclusion] The established HPLC-CAD fingerprint and multi-component content determination method are accurate and stable，and can effectively evaluate the quality of Siegesbeckiae Herba.