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RP-HPLC法同时测定原花青素中儿茶素、表儿茶素、没食子酸、原花青素B2的含量

Determination of catechin, epicatechin, gallic acid and procyanidin B2 in procyanidins by RP-HPLC

  • 摘要: 建立同时测定原花青素中儿茶素(C)、表儿茶素(EC)、没食子酸(GA)、原花青素B2(PCB2)含量的方法,比较不同厂家的原花青素中这4种物质含量的差异。采用RP-HPLC法,利用Waters高效液相色谱仪,色谱柱为Hypersil ODS2(4.0 mm×200 mm,5 μm),以2%冰醋酸溶液和甲醇为流动相,梯度洗脱,检测波长280 nm。结果显示:C、EC、GA和PCB2的质量浓度分别在0.1~50 μg/mL(r=0.998 6),0.1~50 μg/mL(r=0.994 5),0.05~50 μg/mL(r=0.999 9)和0.1~50 μg/mL(r=0.992 2)范围内与峰面积呈良好的线性关系;C、EC、GA、PCB2的平均加样回收率(%)分别为98.36、98.21、89.60、98.47,RSD(%)分别为1.39、0.84、2.12、2.46。该方法简便、准确、重复性好,可用于同时测定原花青素中C、EC、GA、PCB2的含量,不同厂家不同批号的原花青素中这4种物质的含量差异较显著。

     

    Abstract: To determine catechin(C), epicatechin(EC), gallic acid(GA)and procyanidin B2(PCB2)in procyanidins and compare their contents in procyanidins from different manufacturers, an A RP-HPLC method was developed. The determination was carried out on a Hypersil ODS2 column(4. 0 mm×200 mm, 5 μm). The mobile phase consisted of methanol and 2% acetic acid with gradient elution and the detection wave-length at 280 nm. There was a good linear relationship between concentration and the peak area in the range of 0. 1-50 μg/mL(r=0. 998 6)for catechin, 0. 1-50 μg/mL(r=0. 994 5)for epicatechin, 0. 05-50 μg/mL(r=0. 999 9)for gallic acid and 0. 1-50 μg/mL(r=0. 992 2)for procyanidin B2, respectively. The average recoveries of catechin, epicatechin, gallic acid and PCB2 were 98. 36%, 98. 21%, 89. 60% and 98. 47%, respectively and the RSDs were 1. 39%, 0. 84%, 2. 12% and 2. 46%, respectively. The method is simple, accurate, reproducible and can be used for assay of C, EC, GA, PCB2 in procyanidins. There was a great difference in the content of four substance in procyanidins from different manufacturers.

     

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