山银花、金银花中绿原酸和总黄酮含量及抗氧化活性测定

Determination of chlorogenic acid, total flavones, and anti-oxidant activity of Flos Lonicerae Japonicae and Flos Lonicerae

目的对3个不同产地山银花和金银花中绿原酸、总黄酮含量及抗氧化活性进行分析,并比较之间的差异。方法对绿原酸提取条件进行优化,最佳提取条件为65℃下60%乙醇溶液按料液比1∶20超声提取30 min;总黄酮最佳提取条件为65℃下用60%甲醇溶液按料液比1∶10超声提取30 min。分别采用高效液相色谱法和紫外分光光度法对样品的绿原酸和总黄酮进行含量测定,并用DPPH自由基清除率来检测山银花、金银花的抗氧化活性。结果建立了良好的绿原酸和总黄酮含量分析方法,绿原酸在0.119～1.190 mg/m L内线性关系良好,r2为0.999 2(n=6);总黄酮在8.48～50.88μg/m L内线性关系良好,r2为0.999 5(n=6)。湖南产地山银花绿原酸量、总黄酮量及自由基清除率分别为3.99%、13.43%、62.41%。重庆产地山银花绿原酸量、总黄酮量及自由基清除率分别为3.29%、10.08%、51.48%。广西产地金银花绿原酸量、总黄酮量、自由基清除率分别为2.55%、7.10%、39.51%。结论提出了结合化学成分分析和抗氧化性活性来比较不同产地山银花和金银花之间区别的分析方法,从"谱-效"结合思路,为山银花和金银花质量控制及鉴别提供一种新的模式。

Objective To determine the content of chlorogenic acid, total flavones, and anti-oxidant activity in Flos Lonicerae japonicae and Flos Lonicerae obtained from three different origins and compare their differences. Methods The optimized extraction conditions of chlorogenic acid were ultrasonic extraction 30 min in 65 ℃ with ethanol: water(60︰40) and solid-liquid ratio(1︰20). The optimized extraction conditions of total flavonoids were ultrasonic extraction 30 min at 65 ℃ with 60% methanol solution, solid-liquid ratio(1︰10). HPLC and UV spectrophotometry were used to determine the content of chlorogenic acid and total flavonoids in the samples, and then estimation of anti-oxidative activity of Flos Lonicerae japonicae and Flos Lonicerae by DPPH radical scavenging capacity method. Results A method for the analysis of chlorogenic acid and total flavonoids was established, which have a good linear relationship of chlorogenic acid in 0.119-1.190 mg/mL and R^2 was 0.999 2(n = 6);A good linear relationship between 0.008 and 0.050 mg/mL and r^2 was 0.999 5(n = 6) for analysis of the total flavonoids. The average content of chlorogenic acid, total flavonoids, and free radical scavenging rate of Flos Lonicerae from Hunan Province was 3.99%, 13.43%, and 62.41%, respectively. The average content of chlorogenic acid, total flavonoids and free radical scavenging rate of Flos Lonicerae from Chongqing were 3.29%, 10.08%, and 51.48% respectively. The average content of chlorogenic acid, total flavonoids, and free radical scavenging rate of Flos Lonicerae japonicae from Guangxi Province were 2.55%, 7.10%, and 39.51%, respectively. Conclusion This study proposed an analytical method combining the chemical composition analysis and anti-oxidant activity to compare the differences between the different producing areas of Flos Lonicerae japonicae and Flos Lonicerae. Combining the "spectrum-effect", it provides a new model for the quality control and identification of two plants.