山楂叶多酚的纯化及其抗氧化特性与组分分析

Purification,antioxidant capacity and component analysis of hawthorn leaf polyphenols

探讨山楂叶多酚的纯化工艺,并分析其抗氧化活性和组成成分。比较SP-825,D101,AB-8,XDA-2等四种树脂对山楂叶多酚的吸附性能,筛选最佳树脂,并优化其纯化参数,分析其抗氧化活性、组成成分和红外光谱特性。结果表明,4种树脂中AB-8树脂纯化山楂叶多酚效果较好,其最佳纯化参数为样液初始pH 4.5、初始质量浓度0.064 mg/mL、上样流速1.5 mL/min、洗脱剂乙醇体积分数为70%和洗脱流速为0.6 mL/min,在此条件下,山楂叶多酚的回收率为78.25%,纯化后多酚的纯度提高了4.15倍。山楂叶多酚具有较好的抗氧化能力,其粗提液和纯化液清除DPPH·的IC 50值分别为(1.28±0.14)μg/mL和(0.78±0.09)μg/mL;对螯合Fe 2+的IC 50值分别为(55.58±1.24)μg/mL和(34.13±1.02)μg/mL;总还原能力的IC 50值分别为(3.66±0.18)μg/mL和(2.12±0.23)μg/mL,纯化液清除·OH的IC 50值为(4.49±0.16)μg/mL;表明纯化后其抗氧化能力有显著提高。FTIR分析表明其具有多酚和黄酮的特征峰;HPLC分析表明山楂叶多酚主要含绿原酸、表儿茶素、阿魏酸、芦丁和熊果酸,说明山楂叶多酚具有较高的开发应用价值。

The purification process of hawthorn leaf polyphenols was investigated,and its antioxidant activity and constituents were analyzed.The adsorption properties of selected four macroporous resins(SP-825,D101,AB-8,XDA-2)for hawthorn leaf polyphenols were compared,the optimum purification resins were screened and the purification parameters were optimized.The antioxidant activity,composition and infrared spectroscopic characteristics of hawthorn leaf polyphenols were investigated.The results showed that AB-8 resin possessed the best effect on the purification of hawthorn leaf polyphenols.The optimum purification parameters were as follows,initial sample solution pH 4.5,sample concentration 0.064 mg/mL,sampling flow rate 1.5 mL/min,elution flow rate 0.6 mL/min with 70%ethanol.Under these conditions,the recovery rate was 78.25%,the purity of polyphenol was increased by 4.15 times after purification.The crude extract and purified substance of hawthorn leaf polyphenols showed good antioxidant activity,the IC 50 values of DPPH·scavenging capacity were(1.28±0.14)μg/mL and(0.78±0.09)μg/mL respectively,the IC 50 values of Fe 2+chelating capacity were(55.58±1.24)μg/mL and(34.13±1.02)μg/mL respectively,the IC 50 values of total reduction power were(3.66±0.18)μg/mL and(2.12±0.23)μg/mL respectively,the IC 50 values of purified hawthorn leaf polyphenols for·OH scavenging capacity were(4.49±0.16)μg/mL,which indicated that the antioxidant capacity of the purified substance of hawthorn leaf polyphenols was significantly improved compared with the crude extracts.FTIR analysis showed that the purified hawthorn leaf polyphenols had characteristic peaks of polyphenols and flavonoids,HPLC analysis showed that the purified hawthorn leaf polyphenols mainly contained chlorogenic acid,epicatechin,ferulic acid,rutin and ursolic acid,which indicated that hawthorn leaf polyphenols have good utilization and development prospects.