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海带中细胞激动素的纯化分离方法
引用本文:黄冰心,范 晓,韩丽君.海带中细胞激动素的纯化分离方法[J].海洋与湖沼,2002,33(5):509-514.
作者姓名:黄冰心  范 晓  韩丽君
作者单位:中国科学院海洋研究所,青岛,266071
基金项目:国家高技术研究发展计划(863)资助项目,2001AA620503号.
摘    要:海带样品于 2 0 0 1年采自青岛太平角海湾 ,用三种不同方法处理样品 ,通过对实验方法及条件的筛选 ,确立了提取、纯化和分离海带中细胞激动素的方法步骤和流程。结果表明 ,最佳方法的基本步骤为 :( 1 )用甲醇∶水∶醋酸 ( 70∶ 30∶ 3,V/V/V)从海带组织中提取出细胞激动素 ,再用PVPP柱去除提取液中的酚类和其他杂质 ;( 2 )将提取液通过连在一起的PVPP柱、DEAE柱和C1 8柱 ,以纯化样品并同时从自由基、核苷、核苷酸和葡糖苷型细胞激动素混合物中分离出核苷酸型细胞激动素 ;( 3)用接有氨基柱的正相液相色谱进一步将葡糖苷型细胞激动素分离出来 ;( 4 )用C1 8液相色谱柱分离其他各种细胞激动素

关 键 词:海带  细胞激动素  纯化  高效液相色谱
收稿时间:2001/10/23 0:00:00
修稿时间:2001年10月23

METHOD OF EXTRACTION AND PURIFICATION OF CYTOKININS IN LAMINARIA JAPONICA
HUANG Bing-Xin,FAN Xiao and HAN Li-Jun.METHOD OF EXTRACTION AND PURIFICATION OF CYTOKININS IN LAMINARIA JAPONICA[J].Oceanologia Et Limnologia Sinica,2002,33(5):509-514.
Authors:HUANG Bing-Xin  FAN Xiao and HAN Li-Jun
Institution:Institute of Oceanology, The Chinese Academy of Sciences;Institute of Oceanology, The Chinese Academy of Sciences;Institute of Oceanology, The Chinese Academy of Sciences
Abstract:This report comprehensively reports the methods of extraction and purification of cytokinins in Laminaria japonica . A series of experiments have been carried out on every key step of the procedure for extraction and purification of cytokinins in Laminaria japonica . Initial extraction of plant material is followed by the removal of phenolics and other impurities using a polyvinylpolypyrrolidone powder (PVPP) columm. To purify the sample and simultaneously achieve the separation of the cytokinin nucleotides from the free base, riboside, and glucoside forms, the sample is passed through columns containing PVPP, DE52 , and C 18 , which are linked in series. Following purification, separation of cytokinin glucosides from the bases and ribosides is achieved by using normal phase HPLC with an amine column. Separation of individual cytokinins then occurs during reverse phase C 18 HPLC.
Keywords:Laminaria japonica      Cytokinins  Purification    HPLC
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