阿勒泰锦鸡儿花及叶中总黄酮的超声提取工艺、抗氧化活性及化学成分研究

Ultrasonic extraction process,antioxidant activity and chemical constituents of total flavonoids from flowers and leaves of caragana altaica

目的研究阿勒泰锦鸡儿中总黄酮的最佳超声提取工艺,对花和叶中总黄酮进行抗氧化研究和成分分析。方法采用响应面试验确定锦鸡儿根、茎、叶和花中总黄酮的最佳提取工艺。采用DPPH自由基、·OH自由基清除作用和铜离子还原能力试验评价阿勒泰锦鸡儿中总黄酮抗氧化能力,通过LC-MS对锦鸡儿叶和花中主要黄酮成分进行鉴定。结果锦鸡儿中总黄酮超声提取最佳工艺为:超声提取时间50 min,料液比1∶50 g/mL,乙醇体积分数60%;花和叶中总黄酮含量分别为22.3、27.3 mg/g。总黄酮浓度为0.04 mg/mL时,花、叶对DPPH自由基清除率分别为84.4%、79.7%;总黄酮浓度为4.0 mg/L时,花、叶对·OH自由基清除率分别为30.5%、27.1%;总黄酮浓度为3.2μg/mL时,花、叶对铜离子总还原百分率为91.8%、85.4%,3种抗氧化试验均显示阿勒泰锦鸡儿花抗氧化能力强于叶。通过LC-MS鉴定表明阿勒泰锦鸡儿叶中有8种黄酮苷,花中有9种黄酮苷,其花和叶中共有的黄酮苷为:槲皮素-3-O-(二鼠李糖)-芸香糖-7-O-鼠李糖苷、山奈酚-3-O-(二鼠李糖)-芸香糖-7-O-鼠李糖苷、槲皮素-3-O-(二鼠李糖)-芸香糖苷、山奈酚-3-O-(二鼠李糖)-芸香糖苷、芦丁、异鼠李素-3-O-芸香糖苷、山奈酚-3-O-芸香糖苷。异鼠李素-3-O-葡萄糖苷只存在于叶中,而杨梅素-3-O-芸香糖苷和异鼠李素-3-O-鼠李糖(1→2)鼠李糖(1→6)葡萄糖苷存在于花中。结论锦鸡儿叶和花中总黄酮含量比根和茎丰富,叶和花中黄酮种类较为相似,由于叶和花中黄酮种类和含量有差异,锦鸡儿花抗氧化活性强于叶。

Objective To study the optimum ultrasonic extraction process of total flavonoids from Caragana altaica,and analyze the antioxidant study and components of total flavonoids in flowers and leaves.Methods The optimum extraction process of total flavonoids from the root stem,leaves and flowers of Caragana.Use of DPPH free radical,·OH free radical and copper ion reduction test to evaluate antioxidant capacity,the main flavonoids in Caragana leaves and flowers were identified by LC-MS.Results The best ultrasonic extraction technology of total flavonoids from Caragana showed:ultrasonic extraction time is50min,solid-liquid ratio is150g/mL,60%ethanol;The content of total flavonoids in the flowers and leaves22.3mg/g,27.3mg/g.When the concentration of flavonoids was0.04mg/mL,the scavenging rates of flowers and leaves on DPPH free radical were84.4%and79.7%;when the concentration of flavonoids was4.0mg/L,the scavenging rates of flowers and leaves on·OH free radical were30.5%and27.1%;when the concentration of flavonoids was3.2μg/mL,the percentage of total reduction of flowers and leaves on copper ion reduction ability were91.8%and85.4%.Three kinds of antioxidant tests showed that the antioxidant ability of flowers was stronger than that of leaves.Being identified by LC-MS,there are8flavonoid glycosides in the leaves,the flowers have9flavonoid glycosides,the flowers and leaves are common in Quercetin-3-O-(two rhamnose)-rutin-7-O-rhamnoglycoside,kaempferol-3-O-(two rhamnose)-rutin-7-O-rhamnoglycoside,quercetin-3-O-(two rhamnose)rutinoside,kaempferol-3-O-(two rhamnose)rutinoside,rutin,isorhamnetin-3-O-rutinoside,kaempferol rutinoside.isorhamnetin-3-O-glucoside is only exists in leaf,and Arbutin-3-O-rue glycoside and isorhamnetin-3-O-rhamnose(1-2)rhamnose(1-6)in flower glucoside.Conclusion The content of total flavone in Caragana is rich in leaves and flowers of the root.The stem leaf and flower are similar in lavones species.The antioxidant activity of Caragana flower is stronger than that in leaves due to the difference in the species and content of Flavonoids from leaves and flowers.