屏边三七多糖的含量测定及抗氧化活性研究

Determination of Polysaccharide Content and Antioxidant Activity of Panax stipuleanatus Tsai

采用可见光分光光度法,在单因素考察多糖显色条件的基础上,采用正交实验设计法确定了多糖的最佳显色条件,并采用苯酚-硫酸法测定屏边三七的多糖含量;通过过氧化氢、1,1-二苯基-2-三硝基苯肼(DPPH)自由基、2,2′-联氮-双-3-乙基苯并噻唑啉-6-磺酸(ABTS+)自由基和羟基自由基的清除能力及总还原力测定多糖的抗氧化活性评价了抗氧化能力。结果表明,最佳显色条件为:质量分数为6%的苯酚1.2 mL、浓硫酸4.5 mL、60℃加热6 min;葡萄糖标准品检测质量浓度为2.0024~52.0624 g/mL,与吸光度呈良好的线性关系(r)=0.9991。屏边三七多糖的平均加样回收率为99.92%,平均质量分数为19.31%。其体外抗氧化能力为:羟自由基>过氧化氢>DPPH>ABTS+自由基。优化后的苯酚-硫酸法测定屏边三七多糖总含量的方法简单、重复性好、灵敏度好、结果准确,屏边三七多糖具有较强的抗氧化能力。

Visible spectrophotometry was used to determine the content of total polysaccharides in Panax stipuleanatus tsai by the phenol-sulfuric acid method,while single factor and L9(34)orthogonal test was employed to optimize the color reaction conditions.The antioxidant activity of polysaccharides in Panax stipuleanatus tsai was first determined by the scavenging capacity for hydrogen peroxide,1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)radical,2,2′-diazo-bis-3-ethylbenzothiazoline-6-sulfonic acid(ABTS+)free radical and hydroxyl radical as well as total reducing power.The results showed that the best color conditions were that heating at 60℃for 6 min with 1.2 mL of phenol(6%)and 4.5 mL of concentrated sulfuric acid.The calibration curve was linear within 2.0024~52.0624 g/mL(r=0.9991)for glucose standard.The average recovery and the content of polysaccharide in Panax stipuleanatus tsai were 99.92%and 19.31%,respectively.Its antioxidant capacity in vitro was in turn of hydroxyl radical>hydrogen peroxide>DPPH>ABTS+free radical.After optimization,the phenol-sulfuric acid method was proved to be simple,reproducible,sensitive and accurate for the determination of polysaccharides in Panax stipuleanatus tsai.