苹果花粉中异鼠李素和绿原酸鉴别及总黄酮测定

目的:建立苹果花粉的定性定量分析方法以应用于质量研究。方法:采用电镜鉴别观察花粉形态;采用薄层色谱法,以甲苯-乙酸乙酯-甲酸(5:4:0.2)为展开剂,鉴别异鼠李素;以乙酸丁酯-甲酸-水(7:2.5:2.5)上层溶液为展开剂,鉴别绿原酸。以芦丁为对照,采用紫外-可见分光光度法测定总黄酮含量,以NaNO₂-Al(NO₃)₃-NaOH体系显色,在500 nm波长处测定。结果:苹果花粉电镜特征明显;薄层鉴别斑点清晰、易于识别、专属性强;芦丁的质量浓度在0.004~0.048 mg·mL^-1范围内与吸光度线性关系良好,r=0.9993,平均回收率(n=9)为100.4%,苹果花粉总黄酮含量在13.56~46.96 mg·g^-1之间。结论:建立的定性、定量分析方法经方法学验证,可有效评价苹果花粉的质量。

Objective: To establish a qualitative and quantitative analysis method for apple pollen quality assay.Methods: The pollen morphology was identified and observed by electron microscope.The isorhmnetin was identified by TLC with toluene-ethyl acetate-formic acid(5:4:0.2).The chlorogenic acid was identified using butyl acetate-formic acid-water(7:2.5:2.5) upper solution as developing solvent.The UV-visible spectrophotometry was adopted to determine the total flavonoids using rutin as control, which was developed with NaNO₂-Al(NO₃)₃-NaOH color system at 500 nm wavelength.Results: The SEM feature of apple pollen was obvious.TLC spots were clear, easy to identify, and specific.The linear range was 0.004-0.048 mg(r=0.999 3), the average recovery(n=9) was 100.4%, and the content of total flavonoids in apple pollen was 13.56-46.96 mg·g^-1.Conclusion: The established qualitative and quantitative analysis method which has been proved effective, can be applied to the quality evaluation of apple pollen.