高效液相色谱法测定秋鼠麴草中总黄酮的含量

目的建立秋鼠麴草提取物中总黄酮含量高效液相色谱的测定方法。方法选择乙醇浓度、料液比和提取时间这3个因素进行正交试验,初步优选出常规热回流法提取总黄酮的最佳工艺。色谱柱为HypersilODS2 C18柱（250 mm×4.6 mm,5μm）;流动相为甲醇∶水=33∶67;紫外检测波长为359 nm;流速1.0 ml/min;柱温35℃;以芦丁为对照计算总黄酮含量;进样量10μl。结果最优的总黄酮提取条件为：60%乙醇溶液、料液比为1∶35、提取时间为60 min。芦丁在0.013 2～0.105 6 mg/ml范围内线性良好,样品平均回收率为98.98%,相对标准偏差（RSD）为1.86%。结论利用高效液相色谱法测定秋鼠麴草热回流提取物中总黄酮含量的方法回收率高、精密度好、重复性好,能为药材鼠麴草的质量评价和控制提供可行方法。

Determination of the total flavonoid of Gnaphafium affine D. Don by high-performance liquid chromatography

Objective To develop a high-performance liquid chromatography （HPLC） method for determination of the amount of total flavonoid from Gnaphaliurn hypoleucum DC. Methods The preliminary optimum extracting conditions of total flavonoid were investigated by using orthogonal experimental design. The experimental factor, such as the percentage of ethanol, the ratio of material to liquid, time of the ethanol refluxing technology was studied. HypersilODS2 C18 （250 mm × 4.6 mm, μm） column of （HPLC） was adopted in the flavonoid determination. The MEOH-water （33：67）was used as mobile phase. The flow rate was 1.0 ml/min. The temperature of HypersilODS2 ClS was 35%. The detective wavelength was 359 nm. The concentration of Rutin was used as control and the injection volume was 101μL. Results The optimal extracting conditions were as follows： 60% ethanol solution, the ratio of material to liquid was 1：35 ; the extracting time was 60 min. Rutin in 0.0132 -0. 1056mg/ml range was obtained good linearity. The average recovery was 98.98% with the RSD of 1.86%. Conclusion It was found that HPLC method could be used for determination the concentration of total flavonoid of hot reflux extraction of Gnaphalium hypoleucum DC. This good repeatability providing a feasible means for method proved to be high recovery, good precision and quality evaluation and control of Herb Gnaphalii.