色谱 ›› 2013, Vol. 31 ›› Issue (3): 260-263.DOI: 10.3724/SP.J.1123.2012.11005

• 研究论文 • 上一篇    下一篇

微囊藻毒素-RR的纯化及鉴定

张永路, 黄夏宁, 肖雯晴, 钟晴, 谷康定*   

  1. 华中科技大学同济医学院公共卫生学院环境医学研究所 教育部环境与健康重点实验室, 湖北 武汉 430030
  • 收稿日期:2012-11-19 修回日期:2013-01-13 出版日期:2013-03-28 发布日期:2013-03-22
  • 基金资助:

    国家自然科学基金项目(20977038).

Purification and identification of microcystin-RR

ZHANG Yonglu, HUANG Xianing, XIAO Wenqing, ZHONG Qing, GU Kangding*   

  1. Key Laboratory of Environment and Health of Ministry of Education, Institute of Environmental Medicine, School of Public Health, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
  • Received:2012-11-19 Revised:2013-01-13 Online:2013-03-28 Published:2013-03-22

摘要: 将固相萃取和凝胶色谱技术相结合,建立了以野外采集的水华蓝藻为原料提取和纯化微囊藻毒素-RR(MC-RR)的有效方法。用70%的甲醇溶液溶解35 g藻浆,经离心等系列处理,得粗提液,旋转蒸发去除甲醇;粗提液经HLB柱固相萃取后,得到7.5 mL洗脱液,然后将其浓缩至2 mL,再用Sephadex LH-20凝胶色谱柱分离纯化,洗脱液分管收集,用紫外分光光度计测定每管洗脱液在238 nm的吸收值,并绘制洗脱曲线。使用高效液相色谱对峰值组分进行鉴定,同时利用紫外分光光度计对毒素的光谱特征进行鉴定。最终获得3.65 mg纯度超过90%的MC-RR样品,产品得率为74.1%。其紫外吸收光谱在238 nm处有特征吸收,证明所纯化的样品为MC-RR。

关键词: 纯化, 固相萃取, 凝胶色谱, 微囊藻毒素-RR(MC-RR)

Abstract: An effective method based on the combination of solid phase extraction and gel chromatography to extract and purify microcystin-RR (MC-RR) from natural cyanobacteria bloom was developed. Thirty five gram natural cyanobacterial bloom scum was extracted with 70% aqueous methanol. After the methanol in the crude extract was removed by a rotary evaporator, the solid phase extraction was used for preliminary purification and concentration. And then 7.5 mL eluent was concentrated to 2 mL and loaded onto a Sephadex LH-20 gel chromatographic column for further purification. The eluate was collected by tubes and detected at 238 nm. The elution curve was plotted by tube numbers as X-axis and the absorbance at 238 nm as Y-axis. The purity and the spectral characteristics of the final extract were identified with high performance liquid chromatograph (HPLC) and spectrophotometer, respectively. A total of 3.65 mg of MC-RR with a final yield of 74.1% was obtained, and the purity was more than 90% with characteristic UV absorption spectra at 238 nm.

Key words: gel chromatography, microcystin-RR (MC-RR), purification, solid phase extraction