油菜蜂花粉中黄酮类化合物的提取与鉴定

Extraction and Identification of Flavonoids in Rape Bee Pollen

以湖北大宗油菜蜂花粉为材料,对其中微量特征成分黄酮类化合物进行鉴定。采用超声波辅助技术提取,运用二次回归正交旋转组合实验优化提取工艺,通过响应面分析和岭嵴寻优分析,确定黄酮类化合物的最佳提取条件;采用HPLC-DAD以及HPLC-MS对酸解后的提取液进行分析。结果表明:超声波辅助技术提取油菜蜂花粉中黄酮类化合物的最佳条件为体积分数76%乙醇、料液比1:30、超声波时间33min,超声波温度为51℃。蜂花粉中总黄酮含量的理论得率为2.12%,实际得率为1.89%,实测得率达到理论预测值的89.15%。HPLC-DAD以及HPLC-MS分析结果证实油菜蜂花粉中可能含有9种黄酮苷元。其中可以推定的黄酮类物质是槲皮素、山奈酚、异鼠李素和柚皮素。可为识别蜂花粉的植物来源,鉴别真伪提供研究方法和实验依据。

Rape bee pollen was used as the raw material to identify its characteristic component-flavonoids, The optimal ultrasound-assisted extraction processing conditions for flavonoids from rape bee pollen were explored by quadratic regression orthogonal rotation experiments, response surface analysis and ridge analysis. The extracted sample with acid hydrolysis was analyzed using HPLC-DAD and HPLC/MS. Results showed that the optimal ultrasound-assisted extraction conditions were ethanol concentration of 76%, material-liquid ratio of 1：30, ultrasonic treatment time of 33 min, ultrasonic treatment temperature of 51 ℃. Under the optimal conditions, the yield of flavonoids was 1.89%, which was close to theoretical yield of 2.12%. HPLC- DAD and HPLC/MS analyses exhibited that nine kinds of flavonoid aglycones were identified in rape bee pollen. The identified flavonoids included quercetin, kaempferot, isorharnnetin and naringenin. These investigations provide a research method and theoretical reference for the identification of plant sources and authenticity of bee pollen.