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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。
OBJECTIVE To establish a HPLC method for the determination of total flavonoids in the extract of Radix Aconiti Kusnezoffii. Methods Three factors of ethanol concentration, solid-liquid ratio and extraction time were selected for orthogonal experiment. The optimal extraction process of total flavonoids by conventional hot reflux method was preliminarily optimized. The column was Hypersil ODS2 C18 (250 mm × 4.6 mm, 5 μm). The mobile phase was methanol: water = 33:67. The UV detection wavelength was 359 nm. The flow rate was 1.0 ml / min. The column temperature was 35 ℃. Calculation of total flavonoids; injection volume 10μl. The optimum conditions for the extraction of total flavonoids: 60% ethanol solution, solid-liquid ratio of 1:35, extraction time was 60 min. Rutin showed good linearity in the range of 0.013 2-0.105 6 mg / ml. The average recovery of rutin was 98.98% and the relative standard deviation (RSD) was 1.86%. Conclusions The method of HPLC for the determination of total flavonoids in the extract of Cirrhudicum korshinskii has high recovery rate, good precision and good reproducibility. It can provide a feasible method for the quality evaluation and control of the Chinese kennel.