苹果树皮中12种酚类化合物的HPLC测定方法

Determination method of 12 phenolic compounds in apple bark by HPLC

【目的】为了建立高效液相色谱法(HPLC)同时测定苹果树皮中没食子酸、儿茶酚、儿茶素、绿原酸、咖啡酸、表儿茶素、香豆酸、阿魏酸、芦丁、根皮苷、槲皮素、根皮素12种酚类化合物的方法,【方法】选用Eclipse XDB C18色谱柱(150 mm×4.6 mm i.d,5μm),以甲醇和1%乙酸溶液作为流动相进行梯度洗脱,利用VWD检测器,在各种酚类物质的最优吸收波长下进行检测,【结果】得到12种酚类化合物的线性回归方程,相关系数均在0.999以上,检出限(3RSN)分别为0.02、0.02、0.03、0.01、0.02、0.03、0.01、0.01、0.03、0.03、0.05和0.01 mg.L-1。色谱测定方法和包含样品处理方法的加标回收率分别在56.54%～98.83%和90.95%～100.90%。【结论】整个提取和分析测定流程简便、快速、准确,适合苹果树皮中酚类化合物的分析测定。

[Objective]A high performance liquid chromatographic （HPLC） method for the simultaneous determination of 12 phenolic compounds in apple bark was developed, including gallic acid, catechol, catechin, chlorogenic acid, caffeic acid, epicatechin, coumaric acid, ferulic acid, rutin, phlorizin, quercetin and phloretin. [Method]The separation was performed on an Eclipse XDB C18 column （150 mm x4.6 mm i.d., 5 μm） with linear gradient elution composed of methanol and 1% acetate acid solution. UV- detections were set at wavelengths of absorption maxima of each phenolic compound except coumarie acid by using VWD detector. [Resuh]Linearity ranges of the working curves were tested and their linear re- gression equations, with correlation coefficients of 0.999 were derived. Detection limits （3RSN） of the 12 com- pounds found were 0.02, 0.02, 0.03, 0.01, 0.02, 0.03, 0.01, 0.01, 0.03, 0.03, 0.05 and 0.01 mg·L^（-1）, respectively. The spiking recoveries of HPLC method and analysis method containing sample treatment were in the range of 56.54% to 98.83% and 90.95% to 100.90%, respectively. [Conclusion]It was proved that this procedure is simple, rapid and accurate for efficient determination of phenolic compounds in apple bark.