HPLC法测定溪黄草不同部位中5个二萜的含量

Simultaneous Determination of Five Diterpenoids in Different Parts of Herba Rabdosiae Serrae by Reverse Phase HPLC

目的建立反相高效液相层析（RP-HPLC）法同时测定溪黄草不同部位中5个对映-贝壳杉烷型二萜类延命素[enmein,（1）],黄花香茶菜甲素[sculponeatin A,（2）],诺多星[nodosin,（3）],毛果青茶菜素[isodocarpin,（4）]和毛栲利素[lasiokaurin,（5）]含量的方法,并进行多批次药材成分分析,为溪黄草质量评价提供依据。方法采用Kromasil C18色谱柱（4.6 mm×250 mm,5μm）,流动相为甲醇-0.1%甲酸,梯度洗脱,流速0.7 m L·min-1,检测波长254 nm。结果 5个二萜的标准曲线及线性范围依次为：（1）Y=5246.4X＋13.86（0.075~1.50μg,r=0.9999）;（2）Y=4673.4X＋78.09（0.121~2.43μg,r=0.9999）;（3）Y=3459.4X＋22.33（0.372~7.43μg,r=1）;（4）Y=4432.8X＋14.71（0.144~2.88μg,r=1）;（5）Y=2434.1X＋0.69（0.010~1.960μg,r=0.9999）。平均加样回收率为99.14%~100.7%,RSD为0.48%~2.53%。结论经过系统的方法学考察,该法简便,准确,重复性好,可用于溪黄草药材的质量分析及控制。

Objective To develop a reverse phase HPLC method for the simultaneous determination of five ent-kauranes diterpenoids of enmein（1）,sculponeatinA（2）,nodosin（3）,isodocarpin（4）and lasiokaurin（5）in Herba Rabdosiae Serrae and to assess their quality. Methods The samples were separated on a Kromasil RP-C18（4.6 mm× 250 mm,5μm）column eluted with methanol and 0.1%formic acid as mobile phase in gradient elution. The flow rate was 0.7 mL·min-1,and the detection wavelength was set at 254 nm. Results The calibration curves and linear range of the above five diterpenoids were determined as （1）Y=5246.4X＋13.86（0.075~1.50 μg,r=0.9999）,（2）Y=4673. 4X＋78.09 （0.121~2.43 μg, r=0.9999）,（3）Y=3459.4X＋22.33（0.372~7.43 μg, r=1）,（4）Y=4432.8X＋14.71 （0.144~2.88μg,r=1）,（5）Y=2434.1X＋0.69（0.010~1.960μg,r=0.9999）,respectively. The average recovery of the analytes were from 99.14 % to 100.7 % and the RSD values were from 0.48 % to 2.53 %. Conclusion The method is simple,accurate and repeatable,and can be used for the quality assessment of Herba Rabdosiae Serrae.