高效液相色谱法测定马铃薯中绿原酸和原儿茶酸
Determination of chlorogenic acid and protocatechuic acid in potatoes by HPLC
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摘要: 目的:建立高效液相色谱法测定马铃薯块茎中绿原酸和原儿茶酸的含量,并测定了两种马铃薯贮藏期间绿原酸、原儿茶酸含量的变化。方法:采用高效液相色谱法梯度洗脱,使用反相C18色谱柱,以1%甲酸水溶液和100%甲醇为流动相,流速为0.8 m L/min,柱温:40℃,在326、280 nm波长处分别对绿原酸和原儿茶酸进行测定。结果:绿原酸、原儿茶酸分别在50~500、200~1000μg/m L范围内与峰面积线性良好,相关系数分别为0.9999、0.9992;精密度RSD均小于0.35%,稳定性RSD均小于1.68%,绿原酸、原儿茶酸重复性RSD分别为2.07%、3.58%,绿原酸、原儿茶酸平均加样回收率分别为98.29%、97.81%,RSD分别为1.46%、1.31%(n=5)。结论:该方法线性范围宽,分离效果好,快速、准确,可用于马铃薯块茎中绿原酸和原儿茶酸含量的测定。Abstract: Objective: To establish a method of detecting the content of chlorogenic acid and protocatechuic acid in potato tuber by High Performance Liquid Chromatography( HPLC),this method was used to determine the change of chlorogenic acid,protocatechuic acid content in two species potatoes during storage.Methods: Gradient elution HPLC was used to determine chlorogenic acid and protocatechuic acid.The samples were separated by reversed- phase C18 chromatographic column using 1% formic acid and 100% methanol as a mixed mobile phase. The detection wavelength was set at 326 nm and 280 nm respectively.The column temperature was set 40 ℃. Results:The calibration curves showed good linear relationship between the peak areas and concentrations was50~500 μg / m L for chlorogenic acid,200 ~ 1000 μg / m L for protocatechuic acid,the related coefficient was 0.9999,0.9992. The RSD of precision were all lower than 0.35%.The RSD of stability were all lower than 1.68%.The RSD of repeatability was 2.07% for chlorogenic acid,3.58% for protocatechuic acid. Standard addition recovery of chlorogenic acid,protocatechuic acid was 98.29%,97.81% with relative standard deviation( RSD) of 1.46%,1.31%( n = 5).Conclusion: The method was rapid,accurate with wind linear range and good separation results,which was suitable for the analysis of chlorogenic acid,protocatechuic acid in potato tuber.
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