HPLC法测定尖叶假龙胆中雏菊叶龙胆酮和去甲基雏菊叶龙胆酮含量

Determination of Bellidifolin and Norswertianolin from Gentianella Acuta by HPLC

目的：建立HPLC法同时测定尖叶假龙胆中雏菊叶龙胆酮和去甲基雏菊叶龙胆酮的含量。方法：色谱柱：Thermo Syncronis C18（4.6 mm×250 mm,5μm）;流动相：0.5%磷酸水（A）-乙腈（B）,梯度洗脱,0～15 min,40%～45%B,15～30 min,45%～60%B;检测波长：254 nm;柱温：30℃;流速：1.0 m L·min-1。结果：雏菊叶龙胆酮和去甲基雏菊叶龙胆酮分别在1.99～79.52μg·m L-1（R2=0.999 9）、2.28～56.90μg·m L-1（R2=0.999 7）范围内线性关系良好;平均加样回收率（n=9）分别为100.01%（RSD=1.50%）、100.65%（RSD=1.14%）。结论：该方法简便、快速、准确、可靠,为尖叶假龙胆的质量控制提供参考。

Objective： To establish the quantitative analysis method for bellidifolin and norswertianolin from Gentianella acuta. Methods： High performance liquid chromatography（HPLC） was performed on a Thermo Syncronis C18（4. 6 mm× 250 mm,5 μm）. The water with 0. 5% phosphoric acid（v/v） was used as an aqueous solvent（A） and CH3 CN as an organic solvent（B） with gradient at a flow rate of 1. 0 m L·min-1. The detection wavelength was 254 nm and the column temperature was set at 30 ℃. Results： The quantitative analysis by HPLC showed that the linear range of bellidifolin and norswertianolin was 1. 99 ～ 79. 52 μg·m L-1（R2= 0. 999 9）,2. 28 ～ 56. 90 μg·m L-1（R2= 0. 999 7）,respectively. The average recovery of samples above was,in order,100. 01%（RSD = 1. 50%） and 100. 65%（RSD = 1. 14%）.Conclusion： The method was simple,rapid,accurate and reliable,which provided a reference for the quality control of Gentianella acuta.