维药蜀葵花的多成分质量分析研究

Quality aanalysis of Marshmallow Flowers based on multi-component determination

目的:建立HPLC法测定蜀葵花药材中芦丁、金丝桃苷、异槲皮苷和槲皮素的含量,以上述4个成分的含量为指标,采用Box-Behnken响应面法考察甲醇体积分数、料液比和提取时间对提取效果的影响,优化维药蜀葵花的提取工艺。方法:HPLC条件:SB-C18色谱柱(250 mm×4.6 mm,5μm),流动相为乙腈(A)-0.2%磷酸水(B),梯度洗脱,流速1.0 mL·min^(-1),柱温35℃,检测波长360 nm;在单因素试验基础上,采用Box-Behnken响应面法,以芦丁、金丝桃苷、槲皮素、异槲皮苷成分含量为指标,优化蜀葵花的提取条件。结果:芦丁、金丝桃苷、异槲皮苷和槲皮素质量浓度在5.025~50.25、10.20~102.0、19.91~199.1、16.85~168.5μg·mL^(-1)范围内呈良好的线性关系(r=0.9991~0.9993),平均加样回收率为91.0%~96.2%;不同产地和批次之间,4个成分的含量相差很大;蜀葵花提取的最佳条件:甲醇体积分数为59%,料液比为1∶50,提取时间为90 min,在此条件下建立的模型预测性与实际值相符。结论:建立的含量测定方法稳定、可靠,重复性好,建立的数学模型精度预测度好,可用于蜀葵花的质量评价及提取工艺条件优化。

Objective:To optimize an HPLC method for determination of rutin,hypericin,isoquercitrin and quercetin in Marshmallow Flower.The contents of the above 4 components were taken as the index,the effects of methanol volume fraction,solid-liquid ratio and extraction time on the extraction efficiency was investigated by Box-Behnken response surface method,and the extraction process was optimized.Methods:HPLC was conducted on SB-C_(18)column(250 mm×4.6 mm,5μm),with the mobile phase comprising of acetonitrile and 0.2%phosphoric acid,with gradient elution at a flow rate of 1.0 mL·min^(-1).The column temperature was 35℃and the detection wavelength was 284 nm.Based on single factor experiment,Box-Behnken response surface method was used to optimize the extraction process of Marshmallow Flower,using rutin,hypericin,isoquercitrin and quercetin as indicators.Results:Rutin,hypericin,isoquercitrin and quercetin exhibited good linearity(0.9991-0.9993)among the ranges of 5.025-50.25,10.20-102.0,19.91-199.1 and 16.85-168.5μg·mL^(-1),respectively.The average recoveries were between 91.0%-96.2%.There were significant differences in component contents between different habitats and batches.The optimum process conditions for Marshmallow Flower were as follows:methanol volume fraction was 59%,solid-liquid ratio was 1∶50,extraction time was 90 min,and the predicted results were similar to the method verification results,and the method was feasible and reliable.Conclusion:The established content determination method was stable,reliable and reproducible,Model established by Box-Behnken response surface methodology has good predictability,which can be used for the quality evaluation and optimizing the extraction process of Marshmallow Flower.