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

Determination of polyphenols in pomegranate peel by reverse-phase HPLC

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1.0m L/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 m L/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.