反相高效液相色谱法测定石榴皮多酚类物质方法的建立及分析

摘要

建立了同时测定石榴皮多酚中绿原酸、表儿茶素、鞣花酸和槲皮素含量的反相高效液相色谱方法。采用色谱柱Hypersil ODS2（250 mm×4�6 mm，5μm），其流动相为乙腈0�4％磷酸溶液（体积比17∶83），流速为1�0 mL／min，检测波长310 nm，柱温为30℃。建立安石榴甙测量方法，色谱柱为 Hypersil ODS2（250 mm ×4�6 mm，5μm）；流动相为甲醇2％冰醋酸（体积比7∶93）；体积流量为1�0 mL／min；检测波长为232 nm；柱温25℃。结果表明：绿原酸、表儿茶素、鞣花酸、槲皮素和安石榴甙在一定浓度范围内与峰面积呈良好的线性关系，其平均回收率分别为100�66％（ RSD＝2�06％）、100�05％（ RSD＝0�58％）、100�05％（ RSD＝0�51％）、99�51％（ RSD＝1�22％）和99�79％（ RSD＝0�52％）。此结果说明反相高效液相色谱可用于测定石榴皮多酚类物质。%We established reverse⁃phase HPLC to separate and determine the contents of chlorogenic acid, epicatechin, ellagic acid and quercetin in pomegranate peel polyphenol. The chromatographic conditions were as follows:Hyspersil ODS2(250 mm×4�6 mm,5 μm) at 30 ℃ with UV detector at 310 nm, mobile phase of acetonitrile⁃0�4% phosphoric acid ( 17 ∶ 83 ) , and flow rate 1�0 mL/min. Simultaneously, We established a method to measure the content of pomegranate glycosides. The chromatographic conditions were as follows:Hyspersil ODS2(250 mm×4�6 mm,5 μm)at 25℃ with UV detector at 232 nm, mobile phase of methanol⁃0�4% glacial acetic acid ( 7∶93 ) , and flow rate 1�0 mL/min�Chlorofenic acid,epicatechin,ellagic acid,quercetin and pomegranate glycosides showed a good linear relationship with peak area in an appropriate concentration range,and the average recoveries were 100�66%( RSD=2�06%) , 100�05%( RSD=0�58%) , 100�05%( RSD=0�51%) , 99�51%( RSD=1�22%) and 99�79%( RSD=0�52%) , respectively. Our findings prove that the reverse⁃phase HPLC developed is suitable to determine polyphenols from pomegrunate.