天然产物研究与开发 ›› 2016, Vol. 28 ›› Issue (8): 1325-1331.doi: 10.16333/j.1001-6880.2016.8.028

• 开发研究 • 上一篇    下一篇

雪松松针总多酚的纯化工艺和抗氧化活性研究

石晓峰1,2*,沈薇1,宁红霞1,2,王斌利2,张莉霞2   

  1. 1 甘肃省医学科学研究院药物研究所,兰州 730050;2甘肃中医大学药学院,兰州 730030
  • 出版日期:2016-08-30 发布日期:2017-01-17

Purification and Antioxidant Activity of Polyphenol from Pine Needles of Cedrus deodara

SHI Xiao-feng1,2*,SHEN Wei1,NING Hong-xia1,2,WANG Bin-li2,ZHANG Li-xia2   

  1. 1 Institute of Materia Medica,Gansu Academy of Medical Science,Lanzhou 730050,China;2Pharmacy School,Gansu University of TCM,Lanzhou 730030,China
  • Online:2016-08-30 Published:2017-01-17

摘要: 通过吸附-解吸附能力考察,从树脂NKA-9、S-8、AB-8、D101、HPD-100及HPD-600中筛选出适合纯化雪松松针总多酚的大孔树脂,并确定纯化工艺参数。结果表明,HPD-600树脂为纯化雪松松针总多酚的优良材料;最佳纯化工艺为:上样量为8 BV、浓度1.5 mg/mL、pH 4.0~5.0,以2 BV/h流速进行动态吸附;以10 BV/h流速的6 BV蒸馏水冲洗除杂后,用3 BV的70%乙醇以4 BV/h的流速进行解吸。在此条件下,多酚的纯度由9.88%提高到34.56%,约为纯化前的3.5倍。利用DPPH和ABTS自由基法对纯化前后雪松松针总多酚的抗氧化活性进行了比较,结果显示,清除DPPH和ABTS自由基能力依次为Vc>纯化后的总多酚提取物>BHT>纯化前的总多酚提取物。通过测定纯化后雪松松针总多酚对野生型秀丽隐杆线虫体内活性氧的水平评价其体内抗氧化能力,结果显示,雪松松针总多酚降低了线虫体内的活性氧水平,10 μg/mL雪松松针总多酚提取物溶液与4000 μM/L的Vc对线虫的体内抗氧化能力相当,表现出良好的抗氧化活性。

关键词: 雪松松针, 总多酚, 大孔树脂, 纯化工艺, 抗氧化性能, DPPH自由基, ABTS自由基, 野生型秀丽隐杆线虫

Abstract: According to adsorption and desorption property of macroporous resins to polyphenol,NKA-9,S-8,AB-8,D101,HPD-100 and HPD-600 resins were compared for purifying total polyphenol from pine needles of Cedrus deodara.The results showed that HPD-600 was the most appropriate resin.The optimal conditions were as follows:the loading amount was 8 BV (bed volume),the concentration of total polyphenol was 1.5 mg/mL with pH 4.0~5.0,the loading flow rate was 2 BV/h for dynamic absorption,the volume and flow rate of elution water was 6 BV and 10 BV/h,respectively.The eluting solvent was 70% alcohol,the volume and flow rate of alcohol was 3 BV and 4 BV/h.Under these conditions,the purity of polyphenol increased from 9.88% (crude samples) to 34.56% (purified samples).With BHT and Vc as two positive controls,comparative study on the antioxidant ability was performed between crude and purified samples by DPPH and ABTS free radical scavenging assays.The results showed that the scavenging activity on DPPH radical and ABTS radical were as follows:Vc> purified samples >BHT> crude samples,and DPPH and ABTS radical scavenging activity of purified samples was equivalent to that of Vc.The level of reactive oxygen species (ROS) in C.elegans wildtype (Bristol N2) were detected for evaluation the effects of purified total polyphenol from pine needles of C.deodara on antioxidant activity in vivo.A significant decrease in the level of ROS in C.elegans was observed,after treatment with purified samples,the effects of 10 μg/mL of purified polyphenol from pine needles of C.deodara on antioxidant activity in vivo was equivalent to that of 4000 μM/L of Vc.

Key words: pine needles of Cedrus deodara, polyphenol, macroporous resin, purification technology, antioxidant ability, DPPH·, ABTS·, Caenorhabditis elegans wildtype (Bristol N2)

中图分类号: 

R284.2