高效液相色谱法测定披针叶黄花中黄酮含量

Determination of Flavonoid Content in Thermopsis Lanceolata by High Performance Liquid Chromatography

目的 以芦丁为标准品,用高效液相色谱法测定披针叶黄花中黄酮的含量。方法 通过单因素实验分别比较用三种常用的提取方法和6个不同溶剂浓度所得提取液的黄酮浓度,选出最佳提取方法为热回流法,最佳溶剂浓度为60%;进一步通过正交实验确定最佳提取工艺条件为浸泡0 h,回流时间3 h,溶剂体积130 mL,乙醇浓度50%。采用C18色谱柱(250 mm×4.6 mm,5μm),以乙腈∶0.4%磷酸水溶液=20∶80进行等度洗脱,流速1.0 mL/min,柱温30℃,紫外检测器检测波长360 nm,测定披针叶黄花中黄酮的含量。结果 芦丁标准溶液在浓度为0.0015~0.0900 g/L范围内线性关系良好,线性回归方程为y=3002 x,R^(2)=0.9996。最优条件下测得2.0000 g披针叶黄花中含有5.4210 mg的黄酮。结论 测定结果具有较好的稳定性和精密度,回收率为99.20%。该方法具有样品处理简单、测定结果准确度高、提取效果好的特点。

Objective This experiment used reeds as the standard to determine the content of flavonoids in conifer yellow flowers by high-efficiency liquid chromatography.Methods By comparing flavonoid concentrations of extracts obtained from three commonly used extraction methods and 6 different solvent concentrations,the best extraction method was selected as thermal reflux method and the optimal solvent concentration was 60%,and the best extraction conditions were 0 h soaking time,3 h reflux and 130 mL 50% ethanol for solvent by orthogonal experiments.Using C18 chromatography column(250 m×4.6 mm,5μm),ethyl acetylene∶0.4% phosphate solution=20∶80 isometric elution,flow rate 1.0 mL/min,column temperature of 30℃,UV detector detection wavelength 360 nm,to determine the content of flavonoids in conifer flavonoids.Results The standard solution of rudin had good linear relations in the concentration range of 0.0015~0.09 g/L,and the linear regression equation was y=3002 x and R^(2)=0.9996.Under optimal conditions,2.0000g of conifer yellow flowers were measured to contain 5.4210 mg of flavonoids.Conclusion The results have good stability and precision,and the recovery rate is 99.20%.The method has the characteristics of simple sample processing,high accuracy of the measurement results and good extraction effect.