罗布麻叶黄酮类成分酶解前后的液相色谱-质谱分析及活性比较  

HPLC-MS Analysis and Activity Evaluation of Flavonoids of Apocynum venetum Leaves before and after Hydrolyzed by Enzyme

作　　者：张语迟[1] 刘春明[1] 刘志强[2] 吴桂梅[1] 李赛男[1] 

机构地区：[1]长春师范学院中心实验室,吉林长春130032 [2]中国科学院长春应用化学研究所绿色化学与过程实验室,吉林长春130022

出　　处：《分析测试学报》2010年第10期1073-1077,共5页Journal of Instrumental Analysis

基　　金：国家自然科学基金资助项目(30970299);吉林省科技厅重点资助项目(20090936)

摘　　要：采用高效液相色谱-电喷雾质谱(HPLC-ESIMS)联用技术,对罗布麻叶中的黄酮类化合物及酶解产物进行了系统的研究。结果表明,罗布麻叶所含黄酮成分极性较大,包含白麻苷、芦丁、山萘酚-3-O-β-D-芸香糖苷、异槲皮素、扁蓄苷和乙酰化金丝桃苷。罗布麻叶黄酮类成分经过β-葡萄糖苷酶和α-鼠李糖苷酶混合酶液酶解后发生水解,水解产物包括金丝桃苷、异槲皮素、乙酰化金丝桃苷、三叶豆苷、紫云英苷、槲皮素和山萘酚。将酶解前后各化合物的高效液相色谱峰面积进行比对,证明白麻苷、山萘酚-3-O-芸香糖苷和扁蓄苷为酶解前罗布麻叶主要成分,金丝桃苷、异槲皮素、扁蓄苷、槲皮素和山萘酚为酶解后的主要成分。细胞实验结果表明,酶解后3个剂量组的细胞相对增殖率较酶解前分别提升了14.61%、22.48%和20.22%。因此,通过β-葡萄糖苷酶和α-鼠李糖苷酶混合酶液酶解可以有效地将罗布麻叶黄酮苷转化为对应的黄酮苷元及疏水性的黄酮苷,大大提高了罗布麻叶黄酮提取物的抗抑郁活性。The major flavonoids and its hydrolysates in Apocynum venetum leave were analyzed by high performance liquid chromatography coupled with electrospray ionization mass spectrometry（HPLC-ESI MS）.The experimental results indicated that the flavonoids extracted from Apocynum venetum leave had the hydrophilic characteristic,which contained quercetin-3-O-β-D-galactosyl-β-D-glucopyranoside,rutin,kaempferol-3-O-β-D-rutinnoside,isoquercetin,avicularin and acetyled hyperoside.After hydrolyzed by complex enzyme of β-glucosidase and α-rhamnosidase,the compounds contained in the extract of raw plant materials of Apocynum venetum were transformed into hydrophobic compounds,e.g.hyperoside,isoquercetin,acetyled hyperoside,astragalin,trifolin,quercetin and kaempferol.The experimental results indicated that quercetin-3-O-β-D-galactosyl-β-D-glucopyranoside,kaempferol-3-O-β-D-rutinnoside and avicularin were the major compounds of Apocynum venetum before hydrolysis,hyperoside,isoquercetin,avicularin,quercetin and kaempferol were the major compounds in hydrolysates.Active evaluation experiment results also showed that the pc 12 cell proliferation of three dose groups after hydrolysis were 68.26%,101.69% and 105.05%,respectively,which raised 14.61%,22.48% and 20.22% respectively,compared with those of three dosage groups before hydrolysis.Therefore,the hydrolysis of β-glucosidase and α-rhamnosidase could be effectively converted flavone glycosides in Apocynum venetum leave into aglycones,and enhanced evidently the anti-depressant ability of flavone glycosides.

关 键 词：高效液相色谱-电喷雾质谱 罗布麻叶 Β-葡萄糖苷酶 α-鼠李糖苷酶 抗抑郁 

分 类 号：O767.72[理学—晶体学] O623.672