微囊藻毒素MCLR的分离与纯化

Separation and purification of microcystin-LR

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.