HPLC法测定三叶青中槲皮素和山奈酚含量研究

目的:建立HPLC法测定三叶青中槲皮素和山奈酚的方法。方法:三叶青组培物样品经甲醇超声提取,HLB固相萃取小柱净化,在色谱条件:色谱柱Varian ODS C18(4.6mm×250mm×5μm)、柱温30℃、流动相0.2%磷酸水-甲醇、流速1.0m L/min、检测波长360nm下进行测定。结果:槲皮素和山奈酚的方法检出限和线性范围分别为0.05mg/kg和0.05~50μg/m L,且线性范围内的相关系数均大于0.999 9,加标回收率为71.9%~105.8%,相对标准偏差为1.5%~3.4%。结论:HPLC法结合固相萃取技术可用于测定三叶青中槲皮素和山奈酚的含量,方法检出限低,结果准确可靠,重复性好。

Study on the Determination of Quercetin and Kaempferol in Radix tetrastigmae by High Perfor-mance Liquid Chromatography

Objective:A high performance liquid chromatography method for determination of quercetin and kaempferol in Radix tetrastigmae was studied. Method:The samples of tissue cultures were extracted with methanol by ultrasonic and purified with HLB solid phase extraction. Then,the analysis was performed on a Varian ODS C18(4.6mm × 250mm × 5μm)chromatographic column with methanol and 0.2% phosphoric acid as mobile phase at a flow-rate of 1.0mL/min,the column temperature was 30℃ and the detection wave?length was 360nm.Results:method detection limits for quercetin and kaempferol were both 0.05mg/kg,and good linearity for them over the range of 0.05~50μg/mL(r>0.999 9). The average recovery and RSD of the method for quercetin and kaempferol were 71.9%~105.8% and 1.5%~3.4%,respectively. Conclusion:high performance liquid chromatography(HPLC)method combined with solid phase extraction technology can be used for the determination of quercetin and kaempferol in Radix tetrastigmae with low method detection limits,high accuracy and good repeatability.