香叶天竺葵组培苗与扦插苗有效活性成分测定及其抗氧化活性研究

Determination of Effective Active Components and Antioxidant Activity in Tissue Culture Plantlets and Seedling Cuttings of Pelargonium graveolens

利用醇提法提取了总黄酮、总酚,通过紫外-可见分光光度法、Folin-酚比色法对组培苗与扦插苗中总黄酮、总酚含量进行了测定比较,并采用DPPH法和羟自由基(·OH)法对抗氧化活性进行了综合评价。研究显示,在香叶天竺葵总黄酮、总酚含量测定方法中,芦丁质量浓度在0~0.03 mg/mL内线性良好(R^(2)=0.9995),平均回收率为97.30%,相对标准偏差为1.29%(n=9);没食子酸在0~0.036 mg/mL内线性良好(R^(2)=0.9991),平均回收率为97.94%,相对标准偏差为0.58%(n=9)。扦插苗总黄酮(0.86%)和组培苗总黄酮(0.911%)含量无显著差异(P>0.05),组培苗中总酚含量为1.124%,显著高于扦插苗总酚含量0.92%(P<0.05);组培苗有效成分对DPPH自由基和·OH半清除能力(IC)分别为16.72和443.92μg/mL,分别是L-抗坏血酸清除能力的33.6%和68.7%;扦插苗的半清除能力(IC)分别为18.86和467.92μg/mL,分别是L-抗坏血酸清除能力的29.8%和65.2%。研究表明,建立的香叶天竺葵总黄酮和总酚含量测定方法操作简单、快捷,具有良好的稳定性、线性、重复性、精密度和回收率。组培苗与扦插苗中总黄酮含量无显著差异,但组培苗总酚含量高于扦插苗含量,组培苗与扦插苗的醇提取对DPPH自由基和·OH具有一定的清除能力,且组培苗的抗氧化活性强于扦插苗,因此可采用组培的方式进行大量繁殖。

Total flavonoids and total phenols in Pelargonium graveolens were extracted by alcohol extraction,and the contents of total flavonoids and total phenols in tissue culture plantlets and seedling cuttings were determined and compared by ultraviolet-visible spectrophotometry and Folin-phenol colorimetric method.The antioxidant activities were evaluated by DPPH method and hydroxyl radical(·OH)method.The results showed that in the method for determining the total flavonoids and total phenols content of Pelargonium graveolens,rutin had sound linearity within the concentration range of 0~0.03 mg/mL(R^(2)=0.9995).The average recovery was 97.30%,and relative standard deviation(RSD)was 1.29%(n=9).Gallic acid had sound linearity within the concentration range of 0~0.036 mg/mL(R^(2)=0.9991).The average recovery was 97.94%,and relative standard deviation(RSD)was 0.58%(n=9).There was no significant difference(P>0.05)in the total flavonoids between seedling cuttings(0.86%)and tissue culture plantlets(0.911%).The total phenolic content in tissue culture plantlets was 1.124%,which was significantly higher(P<0.05)than that in seedling cuttings(0.92%).The semi-scavenging capacity(IC)of effective components of tissue culture plantlets to DPPH free radical and OH was 16.72μg/mL and 443.92μg/mL,which were 33.6%and68.7%of L-ascorbic acid,respectively.The ICof seedling cuttings was 18.86μg/mL and 467.92μg/mL,which were 29.8%and 65.2%of that of L-ascorbic acid,respectively.The established method for determine the content of total flavonoids and total phenols in Pelargonium graveolens eas simple,rapid,and had good stability,linearity,repeatability,precision and recovery rate.There was no significant difference in total flavonoids content between tissue culture plantlets and seedling cuttings,but the total phenols content of tissue culture plantlets was higher than that of seedling cuttings.Alcohol extraction of tissue cultured plantlets and seedling cuttings had certain scavenging ability on DPPH free radicals and·OH,and the antioxidant activity of tissue culture plantlets was stronger than that of seedling cuttings,so tissue culture plantlets could be used for propagation.