UPLC法测定芭蕉药材不同部位中羽扇豆酮和豆甾醇的含量

Content Determination of Lupanone and Stigmasterol in Different Parts of Musa basjoo by UPLC

目的:建立检测同植株芭蕉根茎、茎和叶中羽扇豆酮和豆甾醇含量的方法,并为寻找芭蕉根茎有效成分的替代资源提供依据。方法:采用超高效液相色谱(UPLC)法进行测定,色谱柱为Zorbax Rrhd Eclipse Plus C18(100 mm×2.1 mm,1.8μm),流动相为乙腈-甲醇(78.5∶21.5,V/V),检测波长为210 nm,流速为0.15 mL/min,柱温为30℃,进样量为1μL。对9批同植株芭蕉根茎、茎和叶中羽扇豆酮和豆甾醇的含量测定结果进行组间差异性分析、主成分分析和聚类分析。结果:羽扇豆酮和豆甾醇的质量浓度分别在11.16~357.10、8.83~160.40µg/mL范围内与各自峰面积呈良好的线性关系(R2分别为0.9992、0.9991);精密度、重复性、稳定性试验的RSD均小于3%;平均加样回收率分别为101.44%、98.32%,RSD分别为1.77%、1.81%(n=6)。芭蕉茎中羽扇豆酮和豆甾醇的平均含量显著高于根茎和叶(P<0.05);同植株芭蕉根茎与叶中羽扇豆酮和豆甾醇的含量比较,差异无统计学意义(P>0.05)。主成分分析结果表明,同植株芭蕉根茎、茎、叶中羽扇豆酮和豆甾醇的含量具有差异。聚类分析将芭蕉根茎、茎和叶分为3大类,其中以芭蕉茎与其余2个部位的差异较明显。结论:该方法简单、快捷、专属性强、重复性好、准确性高,可用于芭蕉药材不同部位中羽扇豆酮和豆甾醇的含量测定;芭蕉茎可替代芭蕉根茎作为羽扇豆酮和豆甾醇的原药材来源。

OBJECTIVE:To establish a method for determining the contents of lupenone and stigmasterol in the rhizome,stem and leaf of Mosa basjoo from the same plant,and to provide reference for the substitute resource for the effective components of M.basjoo.METHODS:UPLC method was adopted.The determination was performed on Zorbax Rrhd Eclipse Plus C18 column(100 mm×2.1 mm,1.8μm)with mobile phase consisted of acetonitrile-methanol(78.5∶21.5,V/V).The detection wavelength was set at 210 nm;the flow rate was 0.15 mL/min;the column temperature was 30℃and the sample size was 1μL.The results of content determination of lupinone and stigmasterol in the rhizome,stem and leaf of 9 batches of M.basjoo from the same plant were analyzed by the methods of comparative analysis between groups,principal component analysis and cluster analysis.RESULTS:The mass concentration of lupenone and stigmasterol had a good linear relationship with the corresponding peak area within the range of 11.16-357.10 and 8.83-160.40 g/mL(R2 were 0.9992 and 0.9991,respectively).RSDs of precision,repeatability and stability tests were all less than 3%.The average recovery rates of lupenone and stigmasterol were 101.44%and 98.32%,and the RSDs were 1.77%and 1.81%(n=6),respectively.The average contents of lupenone and stigmasterol in stems of M.Basjoo were significantly higher than those of rhizome and leaves of M.basjoo(P<0.05).There was no statistical significance in the contents of lupenone and stigmasterol between stem and leaf of M.basjoo from same plant(P>0.05).Results of principal component analysis showed that the contents of lupanone and stigmasterol were different in rhizome,stem and leaf of M.basjoo from the same plant.Rhizome,stem and leaf of M.basjoo were divided into three types through cluster analysis,among which the rhizome had significant difference with the other two parts.CONCLUSIONS:The method is simple,rapid,specific,reproducible and accurate.It can be used for the content determination of lupenone and stigmasterol in different parts of M.basjoo.The stem of M.basjoo can replace the rhizome of M.basjoo as the source of lupinone and stigmasterol.