HPLC结合响应面法优化柳叶蜡梅总黄酮提取工艺及其抑菌活性研究

采用高效液相色谱法测定柳叶蜡梅中黄酮类成分芦丁、槲皮素、山奈酚的含量,在单因素考察基础上,采用响应面法研究乙醇浓度、提取时间、提取温度对柳叶蜡梅黄酮类成分得率的影响,并采用滤纸片法对优化提取的柳叶蜡梅黄酮成分进行抑菌效果研究。结果表明,影响黄酮含量的因素为乙醇浓度>提取时间>提取温度;最佳提取工艺为提取温度90 ℃,料液比1:15,乙醇浓度79.0%,提取时间1.7 h;柳叶蜡梅中黄酮类成分为芦丁0.861 mg·g^-1,槲皮素0.504 mg·g^-1,山奈酚0.492 mg·g^-1,总提取量为1.857 mg·g^-1,与预测值的偏差为1.50%。优化提取的总黄酮对多种病原菌有抑制作用,对蜡样芽孢杆菌和大肠埃希菌有较强的抑制作用,抑菌圈直径分别为13.21、11.18 mm。可见,响应面优化柳叶蜡梅总黄酮提取工艺方法可行,提取的总黄酮具较好的抑菌效果,具有良好的开发前景。

Optimization of extraction process of total flavonoids from Chimonanthus salicifolius S.Y.H by HPLC combined with response surface methodology and its antibacterial activity

The contents of rutin, quercetin and kaempferol extracted from Chimonanthus salicifolius were determined by high performance liquid chromatography (HPLC). Based on the single factor analysis, the effect of ethanol concentration, extraction time and extraction temperature on the yield of flavonoids from C. salicifolius were studied by response surface methodology. Meanwhile, the antibacterial activity of flavonoids from C. salicifolius was assessed by filter paper method. The results showed that main factors affecting the extraction of total flavonoids from C. salicifolius were ethanol concentration, extraction time, extraction temperature. The optimum extraction conditions of total flavonoids were: extraction temperature of 90 ℃, solid-liquid ratio of 1:15, ethanol concentration of 79%, extraction time of 1.7 h. Under the optimal conditions, contents of rutin, quercetin, kaempferol were 0.861, 0.504, 0.492 mg·g^-1, respectively; and the extraction yield of total flavonoids was 1.857 mg·g^-1 which differed from the theoretical value of 1.50%,the total flavonoids extracted from Chimonanthus salicifolius had antibacterial activity against many kinds of bacteria, especially Bacillus cereus and Escherichia coli, the mean diameters of the inhibition zones about them were 13.21 and 11.18 mm, respectively. Therefore, response surface methodology was feasible for the extraction optimization of total flavonoids from C. salicifolius. The total flavonoids from C. salicifolius had good antibacterial activity, indicating that it might have great application prospects in the fields of medicine.