马鞭草中总黄酮测定方法优化及其对DPPH自由基清除动力学过程  

An optimized experimental designation for total flavonoid contents determination from Herba Verbenae Officinalis extracts and its DPPH radical scavenging activity by dynamic approach

作　　者：王旭[1,2] 刘妍如[2,3] 杨建云[2] 肖炳坤[2] 甄蓓[2] 董俊兴[2] 黄荣清[2] 

机构地区：[1]安徽医科大学研究生学院,安徽合肥230032 [2]中国人民解放军军事医学科学院放射与辐射医学研究所,北京100850 [3]沈阳药科大学制药工程学院,辽宁沈阳110016

出　　处：《时珍国医国药》2013年第7期1543-1546,共4页Lishizhen Medicine and Materia Medica Research

基　　金：国家自然科学基金(No.81072613);十二五国家科技重大专项(No.2012ZX09301003-001-010)

摘　　要：目的以响应曲面实验优化马鞭草中总黄酮的光谱测定条件,对马鞭草清除DPPH自由基活性进行动力学评价。方法采用Plackett-Barman设计及响应曲面法优化马鞭草中总黄酮的测定条件并进行方法学验证。动力学方法测定0～600 s内总黄酮对DPPH自由基的清除过程。总黄酮测定的最优显色条件为:水浴温度:50℃,水浴时间:25 min,超声时间:30 min。方法优化后,在0.002～0.002 4 mg/ml范围内线性关系良好(R2=0.999 8)。精密度RSD=0.6%,加样回收率RSD=3.14%。结果与对照相比,马鞭草对DPPH自由基具有缓慢而持续的清除活性。结论以响应曲面优化设计对马鞭草提取物中总黄酮的定量分析优化的方法准确、可靠、快速。Objective This paper aims to establish a robust UV approach for the determination of total flavonoid and total phenolic content from Herba Verbenae Officinalis extracts that performing as antioxidant components via an optimized experiment method.Meanwhile,an optimization strategy for the reaction of total flavonoid by UV is presented,with the aid for response surface methodology（RSM）.Methods A Plackett-Barman experimental design was used to screen the main effects of ultrasound frequency,ultrasound duration,the water bathing temperature,the water bathing duration,and the placing-time for coloration on absorbance and concentration.ANOVA result showed that ultrasound duration,the water bathing temperature and the water bathing duration were statistically significant（P 0.05） factors affecting the absorbance and concentration.Then the Box-Behnken experimental design with response surface methodology was applied to optimized the model and the best assay conditions were： ultrasound duration is 30 min,the water bathing temperature is at 50℃,the water bathing duration is 25 min.The optimized method were further validated according to ICH guidelines with linearity and range precision accuracy and stability.At last,the DPPH radical scavenging activity of Herba Verbenae Officinalis extracts was tested via dynamic approach.Results According to the optimized methods,standard calibration carves were prepared with seven calibrators over a concentration range of 0.002 ～ 0.002 4 mg / ml for rutin.The correlation coefficients（R2） were 0.999 8.The precision and accuracy RSD（%） of the method were 0.6% and 3.14% respectively that met the proposed requirement.The total flavonoid contents of Herba Verbenae Officinalis extracts in triplicate,were 18.8,19.4,19.5 mg RE / g.Conclusion This study indicated that the design of experimental method could be effectively,applied to optimize the total flavonoid assay conditions.

关 键 词：马鞭草提取物 总黄酮 Plackett-Barman设计 响应曲面设计 动力学 

分 类 号：R284.1[医药卫生—中药学]