蒙药材悬钩子木特征图谱研究及6种成分含量测定

Study on Characteristic Chromatogram of Mongolian Medicine Rubus Sachalinensis Leveile and Content Determination of 6 Components

关键词:  悬钩子木  特征图谱  含量测定  HPLC-Q-Exactive-MS/MS

Key words:  Rubus sachalinensis Leveile  characteristic chromatogram  content determination  HPLC-Q-Exactive-MS/MS

摘要:

目的 建立11批不同产地悬钩子木特征图谱,并测定6种主要成分含量。方法 采用CAPCELL PAK MGII C₁₈色谱柱(4.6 mm×250 mm,5 μm),以乙腈(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱,检测波长254 nm;建立不同产地的悬钩子木特征图谱,通过HPLC-Q-Exactive-MS/MS进行共有峰的指认,进行相似度评价,并测定6种主要成分的含量。结果 悬钩子木特征图谱共标定24个共有峰,指认了其中18个峰。S1~S8批次悬钩子木指纹图谱与对照指纹图谱的相似度均>0.900,其余3批产地较远,相似度差异较大。经方法学考察,6种成分在一定浓度范围内呈良好的线性关系;精密度、稳定性、重复性均符合要求;平均加样回收率为97.72%~98.94%。其中特征性成分鞣花酸含量较大,且产地分布均匀,可作为质量控制指标性成分。结论 本研究建立的悬钩子木HPLC特征图谱、定量测定分析方法和HPLC-Q-Exactive-MS/MS定性鉴定方法简便、准确、可靠,可为悬钩子木的质量控制提供参考。

Abstract:

OBJECTIVE To establish the characteristic chromatogram of 11 batches of Rubus sachalinensis Leveile from different producing areas and determine the contents of 6 main components. METHODS Used the CAPCELL PAK MGII C₁₈ column(4.6 mm×250 mm, 5 μm), acetonitrile(A)-0.1% formic acid aqueous solution(B) as mobile phase for gradient elution, detection wavelength of 254 nm. Established the characteristic chromatogram of Rubus sachalinensis Leveile from different places. The common peaks were identified by HPLC-Q-Exactive-MS/MS, and the similarity was evaluated, the content of 6 main components was determined. RESULTS A total of 24 common peaks were demarcated and 18 of them were identified. The similarity between the fingerprint of S1-S8 batches of Rubus sachalinensis Leveile and the control fingerprint was >0.900, and there was a significant difference in similarity among the other three batches with distant origins. Through methodological investigation, the six components showed a good linear relationship within certain concentration range. Precision, stability and repeatability met the requirements. The average recoveries were 97.72%-98.94%. The content of characteristic component (ellagic acid) was high, and the origin distribution was uniform, which could be used as the quality control index ingredient. CONCLUSION In this study, HPLC characteristic chromatogram, quantitative determination and analysis method and HPLC-Q-Exactive-MS/MS qualitative identification method of Rubus sachalinensis Leveile are established, which are simple, accurate and reliable, and can provide reference for the quality control of Rubus sachalinensis Leveile.