不同提取方式对萱草花中酚类物质及抗氧化活性的影响

Effects of different extraction methods on phenolic compounds and antioxidant activity in Hemerocallis flower

为了研究超声、酶解-超声以及发酵-超声3种不同提取方式对萱草花中酚类物质的含量、组成形态及抗氧化活性的影响,以明黄、桔黄、桔红、朱红、大红5种颜色的萱草花为原料,检测不同萱草花中总黄酮和总多酚含量以及1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)和2,2'-联氮-双-3-乙基苯并噻唑啉-6-磺酸(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid,ABTS)自由基清除率;利用高效液相色谱对游离态、共轭态和结合态3种形态酚类化合物进行测定,并研究其与抗氧化活性的关系。结果表明,不同颜色的萱草花中总黄酮和总多酚含量各不相同,其含量从高到低排列为:“红鹦鹉”、“63#”、“健壮力兰”、“玫瑰犀牛”、“太阳舞”。不同品种和不同提取方式的萱草花抗氧化性均存在显著(P<0.05)差异,发酵-超声提取方式下“红鹦鹉”、“63#”、“健壮力兰”、“玫瑰犀牛”、“太阳舞”对DPPH自由基清除率的维生素C当量分别为44.32、40.63、38.24、37.64、35.60 mg/g,对ABTS自由基清除率的维生素C当量分别为39.74、36.24、30.88、28.38、24.88 mg/g;“红鹦鹉”对DPPH清除率的维生素C当量由超声时的38.24 mg/g升高到酶解-超声后的41.49 mg/g和发酵-超声后的44.32 mg/g;对ABTS自由基清除率的维生素C当量由超声时的31.82 mg/g上升到发酵-超声后的39.74 mg/g和酶解-超声时的35.12 mg/g;可知发酵-超声提取下萱草花的抗氧化性优于酶解-超声和超声。在萱草花酚类物质的3种形态中,游离态含量最高,占总含量的75%以上;经酶解和发酵后显著(P<0.05)提高了“红鹦鹉”中大部分游离态酚类化合物的含量,新检出了对香豆酸,发酵后游离态槲皮素含量是超声提取时的3倍;游离酚的DPPH和ABTS自由基清除率的维生素C当量分别从超声时的40.53和35.57 mg/g升高到了发酵-超声时的48.20和47.40 mg/g。游离态中的绿原酸、芦丁、咖啡酸和槲皮素与抗氧化活性相关性高,对抗氧化活性贡献大。研究为萱草花在食品和化妆品的开发利用提供了一定的理论基础。

Hemerocallis fulva is one of the most popular flowers rich in bioactive substances,such as polyphenols,flavonoids,and alkaloids anthraquinones,particularly on antioxidant,antidepressant and anticancer.The extraction of phenolic compounds and antioxidant properties can also greatly contribute to the popularization of H.flower in the agricultural industry.However,only a few reports were focused on the extraction,forms and antioxidant activities of phenolic compounds in the H.flower.The present study aims to explore the effects of different extraction on the content,composition and antioxidant activity of phenolic compounds in various Hemerocallis flowers.Five kinds of flowers with different colors(yellow,orange,red-orange,jujube red,and bright red)were selected as raw materials.Three processing treatments(ultrasound,enzymolysis-ultrasound,and fermentation-ultrasound)were also utilized to determine the content of total flavonoids,total polyphenols,and scavenging rate of free radicals.Furthermore,high-performance liquid chromatography was adopted to identify the free,conjugated,and bound fractions of phenolic compounds,where the relationship was also established between phenolic compounds and antioxidant activity.Specifically,the types of total flavonoids and polyphenols were as follows:'Hongyingwu','63#','Jianzhuanglilan','Meiguixiniu',and'Taiyangwu'.The results showed that there was a significant difference(P<0.05)of antioxidant activity in the different varieties of flowers,where the antioxidant activity was positively correlated with phenolic content.Furthermore,the antioxidant activities of flowers depended mainly on the varieties and extraction.More importantly,the vitamin C equivalents of DPPH(1,1-diphenyl-2-picrylhydrazyl)were 44.32,40.63,38.24,37.64,and 35.60 mg/g,respectively,for the'Hongyingwu','63#','Jianzhuanglilan','Meiguixiniu'and'Taiyangwu'under the fermentation-ultrasonic processing.The corresponding ABTS(2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid))vitamin C equivalents were 39.74,36.24,30.88,28.38 and 24.88 mg/g,respectively.The vitamin C equivalent of DPPH of'Hongyingwu'increased from 38.24 mg/g by ultrasonic extraction to 41.49 mg/g by enzymatic ultrasonic extraction and 44.32 mg/g by fermentation-ultrasonic extraction.The vitamin C equivalent of ABTS of'Hongyingwu'increased from 31.82 mg/g by ultrasonic extraction to 35.12 mg/g by enzymatic ultrasonic extraction and 39.74 mg/g by fermentation-ultrasonic extraction.Among the three fractions,the content of phenolic compounds in the free fraction was the highest,accounting for more than 75%of the total contents.Enzymatic hydrolysis and fermentation significantly(P<0.05)improved the contents of most phenolic compounds in the free fraction of'Hongyingwu'.Particularly,the p-Coumaric acid was newly detected,while the free quercetin content after fermentation reached three times that of the original.The Vitamin C equivalent of DPPH and ABTS free radical scavenging rate of free phenols increased from 40.53 and 35.57 mg/g for ultrasonic processing to 48.20 and 47.40 mg/g for fermentation-ultrasonic processing,respectively.Correlation analysis demonstrated that the chlorogenic acid,caffeic acid,quercetin,and rutin in the free fraction were highly correlated with the antioxidant activity,indicating a great contribution to antioxidant activity.The finding can provide a theoretical basis for the development and utilization of H.flower in functional foods and cosmetics.