| 摘要: |
| 海带样品于2001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C18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C18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 |
| DOI： |
| 分类号: |
| 基金项目:国家高技术研究发展计划( 863) 资助项目, 2001AA620503 号 |
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| METHOD OF EXTRACTION AND PURIFICATION OF CYTOKININS IN LAMINARIA JAPONICA |
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HUANG Bing-Xin, FAN Xiao, HAN Li-Jun
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Institute of Oceanology, The Chinese Academy of Sciences
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| 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 C18, 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 C18HPLC. |
| Key words: Laminaria japonica, Cytokinins, Purification, HPLC |
