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目的研究油菜花粉中黄酮类化合物的提取方法并进行提取工艺优化,同时建立HPLC-UV法测定槲皮素、山柰酚和异鼠李素3种黄酮类化合物的方法。方法以总黄酮得率为指标,优选油菜花粉中黄酮类化合物的最佳提取方法和工艺参数;采用Capcell PAK-C18(250 mm×4.6 mm,5μm)色谱柱,乙腈-pH 2.2磷酸水溶液(35∶65)洗脱,检测波长360 nm,体积流量1.00 mL/min,柱温40℃,对提取物中3种黄酮类成分进行分析测定。结果渗漉法为提取油菜花粉中黄酮类化合物的理想方法;3种黄酮类成分分离度良好,各成分质量浓度与峰面积在测定范围内线性关系良好,重现性、稳定性和加样回收率均符合要求。结论渗漉法适合提取油菜花粉中的黄酮类化合物,所建立的HPLC法稳定可靠、简便易行,可用于油菜花粉中黄酮类化合物的同时定量分析,为油菜花粉原料及其提取物的质量控制和评价奠定了基础。
OBJECTIVE To study the extraction method of flavonoids from rape pollen and to optimize the extraction process. At the same time, three flavonoid compounds, quercetin, kaempferol and isorhamnetin were determined by HPLC-UV. Methods The optimal extraction method and process parameters of flavonoids in rape pollen were optimized with the yield of total flavonoids as the index. The chromatographic column of Capcell PAK-C18 (250 mm × 4.6 mm, 5 μm), acetonitrile-pH 2.2 phosphoric acid : 65), the detection wavelength was 360 nm, the volume flow rate was 1.00 mL / min and the column temperature was 40 ℃. The three flavonoids in the extract were analyzed and determined. Results The percolation method was an ideal method for the extraction of flavonoids from rape pollen. The separation of the three flavonoids was good. The linearity of the mass concentration and peak area of each component in the determination range was good. The reproducibility, stability and sample recovery Rate are in line with requirements. Conclusion Percolation method is suitable for the extraction of flavonoids from rape pollen. The established HPLC method is stable and reliable, simple and convenient, and can be used for the simultaneous quantitative analysis of flavonoids in rape pollen, which can be used for quality control of rape pollen raw materials and their extracts And evaluation laid the foundation.