微囊藻毒素MCLR的分离与纯化  

Separation and purification of microcystin-LR

作　　者：王金丽[1,2] 梁文艳[1] 陈莉[1] 

机构地区：[1]北京林业大学环境科学与工程学院 [2]国家城市给水排水工程技术研究中心

出　　处：《北京林业大学学报》2010年第2期184-188,共5页Journal of Beijing Forestry University

基　　金：国家自然科学基金项目(50678024);"863"国家高技术研究发展计划项目(2007AA06Z301)

摘　　要：通过萃取和常压柱层析色谱技术的合理组合,建立了从实验室培养的铜绿微囊藻中提纯微囊藻毒素MCLR的有效方法。MCLR粗提液先经过正己烷萃取脱脂、固相萃取净化,再用Sephadex LH-20凝胶过滤层析色谱分离去除杂质和色度,流出液分管收集,通过测定流出组分在238nm处的吸光度值,并对峰值组分进行鉴定,证明第2个洗脱峰为MCLR,其纯度为40.5%。最后利用DEAE Sepharose CL-6B阴离子交换层析进一步纯化,以水为初始缓冲液,以3mmol/L NaCl水溶液洗脱MCLR,得到MCLR在238nm的特征吸收峰,MCLR的纯度达90%以上,最终得率为66.2%。This study established an effective method to purify microcystin-LR（MCLR） from laboratory-cultured Microcystis aeruginosa by a suitable combination of liquid-liquid extraction and normal-pressure column chromatography technologies.First,the crude extraction of MCLR was processed by n-hexane extraction to remove lipid substances and solid phase extraction（SPE） to realize preliminary purification.Then the Sephadex LH-20 size exclusion chromatography was used to remove pigments and other impurities.The elution from LH-20 was collected by tubes and the absorbance was detected at 238 nm.We discovered that the second curve contained MCLR;its purity reached 40.5% .Finally,the MCLR sample was purified by DEAE Sepharose CL-6B anion exchange column chromatography,in which the ultra-pure water was used as an initial buffer and 3 mmol/L NaCl was used to elute MCLR.The purity of MCLR reached more than 90% ,with a final yield of 66.2% after DEAE.

关 键 词：MCLR 纯化 萃取 柱层析色谱 

分 类 号：X172[环境科学与工程—环境科学]