橘叶黄酮的纯化及其体外抗氧化能力研究
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摘要
优化大孔吸附树脂纯化橘叶总黄酮的工艺条件,对比考察纯化前后橘叶黄酮的体外抗氧化能力。在单因素试验的基础上,采用响应面法设计优化总黄酮吸附工艺,以吸附率为指标,考察上样液质量浓度、上样液pH值、上样液流速3个因素对吸附效果的影响。并对洗脱剂及其用量进行考察。对比研究橘叶黄酮纯化前后清除羟自由基、DPPH自由基的能力。D101型大孔树脂对橘叶总黄酮纯化的较佳工艺为:上样液质量浓度2 mg/mL,上样液流速1 BV/h,上样液pH 2.5;其他条件为上样液总黄酮量与树脂体积之比为7.7∶1(mg/mL),洗脱剂用量与树脂质量之比为5.6∶1(mL/g)。纯化后,总黄酮含量从4.8%提升至72.8%。同一黄酮浓度下,纯化后体外抗氧化能力均较纯化前强。大孔树脂纯化工艺对橘叶黄酮类成分具有良好的富集作用,且纯化后黄酮是总黄酮中发挥抗氧化能力的主要贡献成分。
Purification condition of total flavonoids from citrus leaves by macroporous resin and its antioxidant ability in vitro was studied. Based on the single factor experiment,the response surface method was used to design and optimize,taking the adsorption rate as the index,the effects of 3 factors,such as the concentration of the sample solution,the pH value of the sample solution and the flow rate of the sample solution,on the adsorption effect were investigated. And the eluting agent and its dosage were also investigated. The ability to remove hydroxyl radicals and DPPH free radicals before and after purification of citrus leaves flavonoids was studied. The results showed that the D101 macroporous resin had better purification of total flavonoids from citrus leaves.The extraction of total flavonoids concentration 2 mg/mL,the desorption velocity 1 BV/h,the pH of the sample liquid was 2.5,other conditions are the total flavonoids of the sample liquid:resin volume 7.7∶1(mg/L),the ratio of detergent dosage to resin quality(mL/g)was 5.6∶1. After purification,the total flavone content from 4.8 % to 72.8 %. In the same flavonoid concentration,the antioxidant capacity in vitro was better than that before purification. Macroporous resin purification process has a good enrichment effect on flavonoids in citrus leaves,and the purified flavonoids were the main contributors to the antioxidant capacity in total flavonoids.
Purification condition of total flavonoids from citrus leaves by macroporous resin and its antioxidant ability in vitro was studied. Based on the single factor experiment,the response surface method was used to design and optimize,taking the adsorption rate as the index,the effects of 3 factors,such as the concentration of the sample solution,the pH value of the sample solution and the flow rate of the sample solution,on the adsorption effect were investigated. And the eluting agent and its dosage were also investigated. The ability to remove hydroxyl radicals and DPPH free radicals before and after purification of citrus leaves flavonoids was studied. The results showed that the D101 macroporous resin had better purification of total flavonoids from citrus leaves.The extraction of total flavonoids concentration 2 mg/mL,the desorption velocity 1 BV/h,the pH of the sample liquid was 2.5,other conditions are the total flavonoids of the sample liquid:resin volume 7.7∶1(mg/L),the ratio of detergent dosage to resin quality(mL/g)was 5.6∶1. After purification,the total flavone content from 4.8 % to 72.8 %. In the same flavonoid concentration,the antioxidant capacity in vitro was better than that before purification. Macroporous resin purification process has a good enrichment effect on flavonoids in citrus leaves,and the purified flavonoids were the main contributors to the antioxidant capacity in total flavonoids.
引文
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[14]车冠达,李倩,程宣,等.大孔树脂分离纯化辣椒叶总黄酮的工艺研究[J].现代生物医学进展,2016,16(3):401-409
[15]魏星任,宋晓娟,龙洁仪,等.仙草总黄酮提取纯化工艺优化及其抗氧化活性[J].中成药,2018,40(5):1185-1188
[16]叶松华,冯夏珍,王晓燕,等.Box-behnken设计优化黄刺玫果总黄酮纯化工艺研究[J].中国食品添加剂,2017(10):73-79
[17]陈佳龄,张凯,郭平叶,等.迷迭香、柠檬草、薄荷水提液清除羟自由基能力的研究[J].日用化学品科学,2014,37(3):27-29,37
[2]吴良发,吴燕红,杨明,等.乳块消片中丹参、橘叶与地龙的同板薄层色谱鉴别[J].赣南医学院学报,2015,35(3):350-351
[3]肖培根.新编中药志[M].北京:化学工业出版社,2002
[4]Guihua Cao,Qingrong Fu,Cangman Zhang,et al.Rapid characterization of 96 chemical constituents in Citri Reticulatae Folium(leaves of'Fuju')using HPLC-DAD-ESI-MS[J].Journal of Chinese Pharmaceutical Sciences,2016,25(2):91-110
[5]曹纬国,刘志勤,邵云,等.黄酮类化合物药理作用的研究进展[J].西北植物学报,2003,23(12):2241-2247
[6]Wan P F,Sheng Z L,Han Q,et al.Enrichment and purification of to-tal flavonoids from Flos Populi extracts with macroporous resins andevaluation of antioxidant activities in vitro[J].Journal of Chromatog-raphy B,2014,945:68-74
[7]恽祥惠,邢丽娜,李云,等.建立基于96孔板检测的FRAP法和DPPH法及其在橘核抗氧化活性研究中的应用[J].时珍国医国药,2014,25(5):1275-1278
[8]余兰彬,徐国良,谭志伟,等.DPPH法对5种抗衰老中药抗氧化活性的研究[J].江西中医药,2016,47(8):74-76
[9]汪金玉,帅欧,林励,等.橘叶HPLC指纹图谱研究[J].中药新药与临床药理,2011,22(1):82-85
[10]汪金玉,帅欧,林励,等.橘叶黄酮类成分的研究与薄层色谱鉴别[J].广州中医药大学学报,2011,28(2):191-193
[11]汪金玉,帅欧,林励,等.RP-HPLC法同时测定橘叶中3个黄酮类成分的含量[J].药物分析杂志,2011,31(11):2161-2163
[12]汪金玉,庄满贤,林励,等.正交设计优选橘叶总黄酮提取工艺[J].食品研究与开发,2012,33(4):72-74,84
[13]郝自新,汪玉萍,王和协,等.橘叶药材质量标准研究[J].安徽科技学院学报,2017,31(4):56-61
[14]车冠达,李倩,程宣,等.大孔树脂分离纯化辣椒叶总黄酮的工艺研究[J].现代生物医学进展,2016,16(3):401-409
[15]魏星任,宋晓娟,龙洁仪,等.仙草总黄酮提取纯化工艺优化及其抗氧化活性[J].中成药,2018,40(5):1185-1188
[16]叶松华,冯夏珍,王晓燕,等.Box-behnken设计优化黄刺玫果总黄酮纯化工艺研究[J].中国食品添加剂,2017(10):73-79
[17]陈佳龄,张凯,郭平叶,等.迷迭香、柠檬草、薄荷水提液清除羟自由基能力的研究[J].日用化学品科学,2014,37(3):27-29,37
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