维药驱虫斑鸠菊HPLC-ELSD特征图谱和2个主要成分含量测定

HPLC-ELSD specific chromatogram of Uyghur medicine Fructus Vernoniae and content determination of two main components

目的:采用HPLC-ELSD法建立维药驱虫斑鸠菊药材的特征图谱,并同时测定2个主要成分的含量。方法:采用植化方法分离鉴定12R,13R-二羟基-9-十八碳烯酸和斑鸠菊酸作为对照品,采用Waters Xbridge C18色谱柱(250 mm×4.6 mm,5μm),0.05%三氟乙酸水溶液(A)-乙腈(B)为流动相,梯度洗脱(0~5 min,60%B;5~20 min,60%B→90%B;20~25 min,90%B→92.5%B),流速1.0 mL·min^-1;柱温30℃;ELSD漂移管60℃,载气为N2,流速1.6 L·min^-1。进样量10μL。结果:建立驱虫斑鸠菊特征图谱,标定5个特征峰,25 min内驱虫斑鸠菊的主要色谱峰能够达到完全分离,对15批药材中2个成分进行含量测定。12R,13R-二羟基-9-十八碳烯酸和斑鸠菊酸分别在0.150 8~3.016μg(r=0.999 3,n=6)和0.812 0~16.24μg(r=0.999 2,n=6)范围内线性关系良好,方法的平均回收率(n=6)分别为99.0%(RSD=1.9%)和98.5%(RSD=1.6%);15批驱虫斑鸠菊中上述2个成分含量分别为0.27~1.11%和2.36~6.34%。结论:所建立特征图谱专属性强,结合2个主要成分含量测定能够为驱虫斑鸠菊的质量控制提供科学依据。

Objective:To establish the HPLC-ELSD specific chromatogram and determine two main components of Vernoniae Fructus.Methods:12 R,13 R-dihydroxy-9-octadecenoic acid and vernolic acid were isolated and identified as reference substances by phytochemical methods.HPLC analysis was performed on a Waters Xbridge C18 column(250 mm×4.6 mm,5 μm)with gradient elution of acetonitrile-0.05% trifluoroacetic acid solution at a flow rate of 1.0 mL·min-1(0-5 min,60% B;5-20 min,60%B → 90% B;20-25 min,90%B → 92.5% B).The carrier gas was N2,and the pressure was 1.6 L·min-1.The drift tube in this experiment was set at 60 ℃.The injection volume was 10 μL.Results:HPLC specific chromatogram of Vernoniae Fructus was established and five characteristic peaks were marked.Two characteristic peaks were simultaneously determined by HPLC within 30 min.The linear ranges of 12 R,13 R-dihydroxy-9-octadecenoic acid and vernolic acid were 0.150 8-3.016 μg(r=0.999 3,n=6)and 0.812 0-16.24 μg(r=0.999 2,n=6),respectively.The average recoveries(n=6)of two components were 99.0%(RSD=1.9%),and 98.5%(RSD=1.6%),respectively.The contents of the two components in 15 batch samples of Vernoniae Fructus were 0.27%-1.11% and 2.36%-6.34%,respectively.Conclusion:The HPLC specific chromatogram of Vernoniae Fructus,together with determination of two components showes strong specificity,and it can be used for the quality control of the crude drug.