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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 establish a reverse-phase high-performance liquid chromatography (RP-HPLC) method for the simultaneous determination of five enantiomers of kaempferol-diterpene enmein (1) Methods of content of [sculponeatin A (2)], nodosin (3), isodocarpin (4) and lasiokaurin (5) And analyzed the ingredients of multiple batches of medicinal herbs to provide the basis for the quality evaluation of Methods Kromasil C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase consisted of methanol-0.1% formic acid and eluted with a gradient of 0.7 m L · min-1 at a detection wavelength of 254 nm. Results The standard curve and linear range of five diterpenes were as follows: (1) Y = 5246.4X + 13.86 (0.075-1.50μg, r = 0.9999); (2) Y = 4673.4X + 78.09 = 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); X + 0.69 (0.010 ~ 1.960 μg, r = 0.9999). The average recovery was 99.14% ~ 100.7% with RSD of 0.48% ~ 2.53%. Conclusions After systematical methodological study, the method is simple, accurate and reproducible, and can be used for the quality analysis and control of Xihuangcao medicinal materials.