荷叶黄酮分离鉴定及不同干燥方法对荷叶黄酮及荷叶碱含量影响  

Isolation and identification of flavonoids from lotus leaf and effects of drying methods on the content of flavonoids and nuciferine

作　　者：凌智辉 肖蓓[1] 岳芊羽 欧梓轩 谢红旗[1,2] 曾建国 陆英[1,2] LING Zhi-hui;XIAO Bei;YUE Qian-yu;OU Zhi-xuan;XIE Hong-qi;ZHENG Jian-guo;LU Ying(College of Horticulture,Hunan Agricultural University;National Herbal Medicine Production (Hunan) Technology Center,Changsha 410128,China)

机构地区：[1]湖南农业大学园艺学院 [2]国家中药材生产(湖南)技术中心,长沙410128

出　　处：《天然产物研究与开发》2020年第10期1730-1736,1746,共8页Natural Product Research and Development

基　　金：现代农业产业技术体系建设专项(CARS-21)。

摘　　要：首先采用高速逆流色谱及制备液相色谱分离荷叶中3个主要黄酮类化合物,然后采用高效液相色谱法对不同干燥方法及不同采收时期荷叶样品中该3个黄酮及荷叶碱含量进行测定。以正己烷∶乙酸乙酯∶正丁醇∶甲醇∶0.2%冰乙酸水=1∶3∶2∶1∶4(V∶V)为两相溶剂体系分离荷叶黄酮提取物,得到两个流分,其中A流分为纯度98.78%的化合物3,流分B再经制备液相进行分离,得到纯度为98.36%的化合物1和纯度为96.68%的化合物2。化合物1、2、3经质谱和核磁共振鉴定分别为金丝桃苷、异槲皮苷和槲皮素-3-O-葡萄糖醛酸苷。荷叶中黄酮类化合物含量高低为:槲皮素-3-O-葡萄糖醛酸苷>金丝桃苷>异槲皮苷;60℃烘干样品中三个黄酮及总量显著高于阴干和烘干,荷叶碱含量无显著差异;不同烘干温度处理样中,随着烘干温度升高,各组分含量增大,100℃最佳;不切制整片干燥样品中黄酮总量显著高于切制干燥,荷叶碱无影响;7月到9月,荷叶黄酮总量趋于稳定,荷叶碱呈上升趋势,因此8月和9月为荷叶较佳采收时间。A method of high speed countercurrent chromatography(HSCCC)and liquid chromatography was developed for preparation and separation of three major flavonoids from lotus leaf at first.Then,HPLC was used to determine the contents of flavonoids and nuciferine in lotus leaf with different drying methods and harvesting time.Using n-hexane:ethyl acetate:n-butyl alcohol:methanol:0.2%water ice acetic acid=1∶3∶2∶1∶4(V/V)as two-phase solvent system,elute A and elute B were isolated and prepared from the flavonoids extract from lotus leaf by high speed counter-current chromatography(HSCCC).Elute A was compound 3 with purity of 98.78%.Elute B was further separated by pre-HPLC to get compound 1 with purity of 98.36%and compound 2 with purity of 96.68%.Compounds 1,2 and 3 were identified as hypericin,isoquercitrin and quercetin-3-O-glucuronide respectively by mass spectrometry and NMR.The content order from high to low was quercetin-3-O-glucuronide>hyperoside>isoquercetin.There was no difference in nuciferine content between three drying methods.The content of flavonoids and total content in 60℃oven-dried samples was significantly higher than that in shade-dried and sun-dried samples,while there was no significant difference in the content of nuciferine.The content of each component increased with the drying temperature,and 100℃was the best.There was a significant difference in the content of flavonoids between drying with slice and drying with whole,while there was no difference in the content of nuciferine.From July to September,the content of flavonoids tend to be stable,and the nuciferine tend to rise,August and September are the best harvest time.

关 键 词：荷叶 荷叶黄酮 分离 荷叶碱 干燥方法 

分 类 号：S609+.2[农业科学—园艺学]