HPLC同时测定甜高粱中10种黄酮类化合物含量及抗氧化活性对比研究

Study on simultaneous determination of 10 flavonoid compounds and a comparative analysis of antioxidant activity in sweet sorghum by HPLC

试验旨在建立同时测定甜高粱不同部位中10种黄酮类化合物含量的高效液相色谱(HPLC)检测方法,并对总黄酮提取物抗氧化能力进行评价。试验以甜高粱陇甜粱2号成熟期籽粒、茎秆及全株为研究对象制备总黄酮提取物,利用HPLC同时测定不同部位总黄酮提取物中的儿茶素、氯化芹菜定、花旗松素、芦丁、槲皮素、木犀草素、柚皮素、高北美圣草素、芹菜素及麦黄酮含量。结果显示,10种黄酮类化合物分别在各自线性范围内与色谱峰峰面积线性关系良好,R^(2)为0.9984~0.9999,平均加样回收率为95.0%~105.8%,RSD为1.03%~2.86%。抗氧化性测定结果表明,甜高粱不同部位总黄酮提取物对羟基自由基、超氧阴离子自由基、1,1-二苯基-2-三硝基苯肼(DPPH)自由基和2,2-联氮-二(3-乙基-苯并噻唑-6-磺酸)二铵盐(ABTS^(+))自由基均具有一定的清除能力,不同浓度下清除率均表现为籽粒>全株>茎秆;籽粒总黄酮提取物浓度为1.00 g/L时,籽粒总黄酮提取物对DPPH自由基的清除率高于VC和2,6-二叔丁基-4-甲基苯酚(BHT)。相关性分析结果表明,不同部位总黄酮提取物含量与不同自由基清除率及总还原能力(FRAP)均呈正相关。研究表明,该试验方法简便,结果准确性较高,可用于甜高粱中黄酮类化合物含量的测定;甜高粱不同部位总黄酮提取物具有抗氧化活性。

HPLC method was developed for the simultaneous determination of 10 flavonoid compounds in different parts of sweet sorghum,and the antioxidant activity of total flavonoid extracts was evaluated.The total flavonoid extracts were prepared from mature seeds,stems,and the whole plant of sweet sorghum cultivar Longtianliang No.2.HPLC was employed to simultaneously determine the content of catechin,apigenin,quercetin,rutin,kaempferol,hesperidin,naringin,hyperoside,luteolin,and maysin content in different parts of total flavonoid extracts.The results showed that the 10 flavonoid compounds exhibited good linearity with peak area within their respective linear ranges,with R^(2)ranging from 0.9984 to 0.9999.The average recovery rates were between 95.0%and 105.8%,and the RSD was in the range of 1.03%to 2.86%.The antioxidant activity assays revealed that the total flavonoid extracts from different parts of sweet sorghum had a certain scavenging capacity against hydroxyl radical,superoxide anions radical,DPPH free radical,and ABTS^(+)free radical,and the scavenging rate at different concentrations was in the order of seeds>whole plant>stem.At a concentration of 1.00 g/L,where the DPPH free radical scavenging rate of seed total flavonoid extract exceeded that of VC and BHT.Correlation analysis indicated a positive correlation between the content of total flavonoid extracts and the scavenging rates of different free radicals,as well as the FRAP.The study demonstrates that the developed HPLC method is simple and provides accurate results,suitable for the determination of flavonoid content in sweet sorghum.The total flavonoid extracts from different parts of sweet sorghum exhibit antioxidant activity.