基于HPLC特征图谱、UPLC-Q-TOF/MS定性及多成分定量的黄芩酒炙前后化学成分变化研究

Comparison of chemical compositions before and after wine-frying of Scutellaria baicalensis based on HPLC characteristic chromatogram,UPLC-Q-TOF/MS qualitative and multi-component quantitative analysis

目的比较黄芩酒炙前后化学成分变化,为建立全面的黄芩饮片质量评价方法提供参考。方法HPLC法建立黄芩片、酒黄芩的特征图谱,并生成对照特征图谱,标定共有峰并进行相似度评价。采用UPLC-Q-TOF/MS定性分析黄芩酒炙前后化学成分,利用UPLC-TQMS对2种饮片中11种黄酮类成分(黄芩苷、黄芩素、汉黄芩苷、汉黄芩素、千层纸素A、千层纸苷、野黄芩苷、芹菜素、高车前素、木犀草苷、白杨素)进行定量分析,并以含量为变量进行多元统计分析。结果建立了黄芩片、酒黄芩的特征图谱,标定9个共有峰,2种饮片相似度均在0.947以上。在黄芩饮片中,共发现50种成分,通过多级质谱数据分析,保留时间匹配,并结合对照品及数据库检索,鉴定了其中44种成分。UPLC-TQMS定量结果显示酒炙后黄芩中黄芩苷、汉黄芩苷等黄酮苷类成分含量略有下降,黄芩素、汉黄芩素等黄酮苷元类成分含量稍有增加。经多元统计学分析,2种饮片有明显的分离趋势,载荷图结果表明黄芩苷、汉黄芩苷、千层纸苷的含量差异可能是引起黄芩酒炙前后化学成分变化的主要因素。结论黄芩酒炙后没有新增或消失成分,但成分含量有所变化。所建立黄芩饮片的定性、定量方法重现性好,分析快速、准确,可用于黄芩饮片酒炙前后的质量评价。

Objective To compare the chemical compositions of Scutellariae Radix(SR)before and after wine-frying,and provide a reference for establishing a comprehensive quality evaluation method for SR decoction pieces.Methods Characteristic chromatogram of SR and wine-processed Scutellariae Radix(wpSR)were established using HPLC,the control characteristic chromatogram maps were generated,the common peaks were calibrated and the similarity was evaluated.Chemical compositions of SR and wpSR were identified by UPLC-Q-TOF/MS.The content profiles of 11 flavonoids(baicalin,baicalein,wogonoside,wogonin,oroxylin A,oroxyloside,scutellarin,apigenin,hispidulin,luteoloside and chrysin)of SR and wpSR was determined by UPLC-TQMS,and PCA was used to analysis the content of 11 flavonoids.Results The characteristic chromatogram of SR and wpSR were established,and nine common peaks were calibrated.The similarity of two pieces were all greater than 0.947.Through the analysis of the multistage tandem mass spectrometry,retention time matching combined with the software of database search and literatures,50 compositions were found and 44 compositions were identified in two pieces.The quantitative results showed that the content of flavonoid glycosides such as baicalin and wogonoside decreased slightly,while the content of flavonoid aglycones such as baicalein and wogonin increased slightly.After multivariate statistical analysis,two pieces were divided into two types.The differences of the content of baicalin,wogonoside and oroxyloside may be the main factors causing the change of chemical compositions of wine-frying of SR.Conclusion Chemical profiles did not change after wine-frying of SR,but the content profiles of some compositions were changed.The established method can provide reproducible,efficient,as well as accurate analysis for Chinese medicinal materials.And it should be an eligible tool for the quality evaluation of SR.