石榴皮多酚分离纯化及对脂肪酸合成酶抑制作用的研究

Purification of Polyphenols from Pomegranate Peel and Their Inhibitory Effect on Fatty Acid Synthase

以总多酚质量分数、吸附率、解吸率为指标,进行石榴皮总多酚的分离纯化研究,并通过反相高效液相色谱(reverse phase high performance liquid chromatography,RP-HPLC)法测定石榴皮多酚中的鞣花酸、安石榴苷、绿原酸、槲皮素和表儿茶素质量分数。同时研究石榴皮多酚纯化物对脂肪酸合成酶的抑制作用。结果表明:石榴皮多酚纯化的最佳条件为采用D101大孔树脂,石榴皮溶液进柱质量浓度为10 mg/mL,流速为2 BV/h,清洗用水5 BV,乙醇洗脱剂的体积分数为70%,用量为5.5 BV;纯化后石榴皮多酚质量分数为71.64%,较纯化前多酚质量分数49.31%有明显提高;纯化后石榴皮多酚中安石榴苷、鞣花酸、绿原酸、槲皮素和表儿茶素质量分数分别为46.91%、9.44%、0.53%、0.75%、0.32%。石榴皮多酚提取物对脂肪酸合成酶的半最大效应浓度(concentration for50%of maximal effect,EC50)为0.72 mg/mL,说明石榴皮多酚对脂肪酸合成酶具有较好的抑制作用。

The purification of polyphenols from the crude aqueous extract of pomegranate peel by macroporous adsorption resin was investigated by evaluating total polyphenol content, adsorption rate and desorption rate as functions of the adsorption and desorption conditions. Meanwhile, the contents of punicalagin, ellagic acid, chlorogenic acid, quercetin and epicatechin were determined by reverse phase high performance liquid chromatography(RP-HPLC). In addition, the inhibitory effect of purified polyphenols on fatty acid synthase was explored. The results showed that D101 macroporous resin was the suitable resin for the purification of polyphenols and the purification process involved the following steps: adjustment of the sample concentration to 10 mg/mL, sample loading at a flow rate of 2 BV/h, column washing with 5 BV of distilled water, and elution with 5.5 BV of 70% ethanol. The purified product contained 71.64%, significantly higher than that(49.31%) reported for the crude extract. The contents of punicalagin, ellagic acid, chlorogenic acid, quercetin and epicatechin in the purified polyphenols were 46.91%, 9.44%, 0.53%, 0.75% and 0.32%, respectively. Moreover, the polyphenols from pomegranate peel had a potent inhibitory effect on fatty acid synthase with an EC50(concentration for 50% of maximal effect) of 0.72 mg/mL.