金叶子二萜类化学成分高效液相色谱-蒸发光散射检测法指纹图谱研究

HPLC-ELSD Fingerprint of Diterpenoids from Craibiodendri Yunnanensis Folium

目的建立测定金叶子药材二萜类化学成分的高效液相色谱-蒸发光散射检测(HPLC-ELSD)法指纹图谱,并对其共有峰进行分离鉴定。方法色谱柱为Ecosil C18柱(250 mm×4.6 mm,5μm),流动相为乙腈-水(梯度洗脱),流速为1.0 mL/min,检测器为ELSD检测器,柱温为30℃。采用硅胶、反向(RP)硅胶C18柱色谱等方法对特征图谱中的共有峰化合物进行分离,应用质谱、核磁共振等方法进行结构鉴定;参照各对照品的色谱行为,对共有峰进行归属和指认。结果11批金叶子药材的相似度在0.926~0.999之间;确定了8个共有色谱峰,并指认了4个色谱峰,分别为craiobioside B,grayanoside B,g rayanoside C,paniculosideⅣ,其中paniculosideⅣ为首次从金叶子中分离得到。结论所建立的金叶子二萜类化学成分高效液相色谱指纹图谱灵敏度高,专属性强,重复性好,可为金叶子药材的质量控制提供参考。

Objective To establish a high-performance liquid chromatography-evaporative light scattering detection(HPLC-ELSD)fingerprint for the determination of diterpenoids from Craibiodendri Yunnanensis Folium,and to isolate and identify their common peak compounds.Methods The chromatographic column was Ecosil C18 column(250 mm×4.6 mm,5μm),the mobile phase was acetonitrile-water(gradient elution),t he flow rate was 1.0 mL/min,the detector was evaporative light scattering detector(ELSD),and the column temperature was 30℃.The common peak compounds in the fingerprints were separated by silica gel and reverse silica gel C18 column chromatography,and their structures were identified by mass spectrometry(MS)and nuclear magnetic resonance(NMR).The common peaks were assigned and identified according to the chromatographic behavior of each reference substance.Results The similarity of 11 batches of Craibiodendri Yunnanensis Folium ranged from 0.926 to 0.999.Eight common chromatographic peaks were found and four chromatographic peaks were identified,which were craioboside B,grayanoside B,grayanoside C and paniculosideⅣ.PaniculosideⅣwas isolated from Craibiodendri Yunnanensis Folium for the first time.Conclusion The established HPLC fingerprint of the diterpenoids from Craibiodendri Yunnanensis Folium has high sensitivity,strong specificity,and good repeatability,which can be used as a reference for the quality control of original medicinal materials of Craibiodendri Yunnanensis Folium.