超临界流体色谱法快速定量分析吴茱萸中的吴茱萸碱和吴茱萸次碱

Rapid Quantification of Evodiamine and Rutae⁃carpine in Evodia Rutaecarpa(Juss.)Benth.Using Supercritical Fluid Chromatography

采用超临界流体色谱(SFC)对吴茱萸中的吴茱萸碱和吴茱萸次碱进行快速定量分析。通过优化后的超临界流体萃取(SFE)条件得到目标生物碱,利用优化后的SFC方法在6 min内完成目标生物碱的分析,并实现了吴茱萸碱与吴茱萸次碱的基线分离。验证结果表明,SFC方法的线性较好,相关系数(r^(2))均为0.9998,精密度良好,相对标准偏差(RSD)均低于0.50%,回收率为102%~109%。吴茱萸碱的检出限和定量下限分别为1.00、3.33μg/mL,吴茱萸次碱的检出限和定量下限分别为0.95、3.17μg/mL。应用该方法检测4个产地的10个吴茱萸样品,目标生物碱的总含量依次为2.95%(广东,1)、0.51%(广东,2)、1.27%(贵州,3)、1.00%(湖南,4)、0.93%(湖南,5)、1.81%(江西,6)、0.73%(江西,7)、0.58%(江西,8)、0.41%(江西,9)和0.36%(江西,10)。尽管含量均符合药典要求,但产地差异显著,且同一产地的药材质量也明显不同。此外,将该方法与2020版中国药典方法进行了比较。两种方法的定量结果相似,但SFC的分析时间明显少于药典方法。研究结果表明SFC在中药活性成分定量方面具有潜力。

This study proved the feasibility of using supercritical fluid chromatography(SFC)in rapid quantification ofevodiamine and rutaecarpine in Evodia rutaecarpa(Juss.)Benth.(E.rutaecarpa).The two alkaloids were firstly extracted by supercritical fluid extraction(SFE).As a result,the developed SFE conditions were as follows,sample(120-200 mesh)/cellulose acetate=1∶1,20% ethanol as modifier,40 ℃,25 MPa and the CO_(2) flow rate of 8 mL/min.The developed SFC method was carried out on a 2-EP column,with methanol as modifier at a flow rate of 1.4 m L/min,a column temperature of 35 ℃ and a back pressure of 15 MPa.The baseline separation of evodiamine andrutaecarpine was completed within 6 min.After verification,the SFC method was confirmed to be linear with correlation coefficients(r^(2))of 0.999 8,good precision(RSD below 0.50%)and accuracy(recoveries of 102%-109%).Meanwhile,limits of detection(LODs)and limits of quantitation(LOQs)for evodiamine were 1.00 and 3.33 μg/m L,while LOD and LOQ for rutaecarpine were 0.95 and 3.17 μg/m L,respectively.Finally,the method was applied tothe determination of contents of evodiamine and rutaecarpine in ten E.rutaecarpa samples from 4 areas.The totalcontent of target alkaloids accounted 2.95%for Guangdong,1,0.51%for Guangdong,2,1.27%for Guizhou,3,1.00%for Hunan,4,0.93%for Hunan,5,1.81%for Jiangxi,6,0.73%for Jiangxi,7,0.58%for Jiangxi,8,0.41%for Jiangxi,9 and 0.36%for Jiangxi,10.Although the ten samples all meet the requirements for the ChinesePharmacopoeia,the origin difference of sample quality is very obvious,and the samples from the same place alsohave differences.In addition,the developed method was compared with the Pharmacopoeia method(2020 Edition).The results of content determination by the two methods were similar(1.27%by SFC vs.1.12%by Pharmacopoeia),while the time required by SFC(6 min)was significantly shorter than that by the Pharmacopoeia method(25 min).This work showed the potential of SFC in the fast quantification of active components in traditional Chinese medicines.