Rundberget Thomas, Sandvik Morten, Larsen Kristofer, Pizarro Gemita M, Reguera Beatriz, Castberg Tonje, Gustad Eli, Loader Jared I, Rise Frode, Wilkins Alistair L, Miles Christopher O
National Veterinary Institute, PB 8156 Dep., NO-0033 Oslo, Norway.
Toxicon. 2007 Dec 1;50(7):960-70. doi: 10.1016/j.toxicon.2007.07.003. Epub 2007 Jul 27.
Marine biotoxins from microalgae can accumulate in shellfish and lead to poisoning of human consumers as well as fish, marine mammals and sea birds. Toxicological assessment of the toxins and development of analytical methods require large amounts of high-purity toxins and their metabolites. Although these toxins can be obtained in limited amounts from contaminated shellfish or from microalgal cultures, difficulties arise when the toxin-producing microalga is difficult to culture or its identity is not known. To circumvent this problem, we have developed a large-scale method for solid-phase extraction (SPE) of lipophilic biotoxins from natural microalgal blooms in seawater. To enhance subsequent purification of toxins adsorbed on the column, we included a filtration step to release the toxins from the cells while removing insoluble compounds and cellular debris. The efficacy of the method was illustrated by extraction and purification of okadaic acid and dinophysistoxin-2 from a high-density Dinophysis acuta bloom in Spain and from a mixed bloom containing low densities of D. acuta in Norway. Isolation of the toxins adsorbed on the SPE column was simple and efficient, and results obtained so far indicate that the method is potentially applicable to a wide range of microalgal toxins such as azaspiracids, pectenotoxins, spirolides and microcystins. The method should also be useful for harvesting toxins from large-scale microalgal cultures, and for bioprospecting for and isolation of bioactive natural products from marine and freshwater environments.
来自微藻的海洋生物毒素可在贝类中蓄积,导致人类消费者以及鱼类、海洋哺乳动物和海鸟中毒。对这些毒素进行毒理学评估以及开发分析方法需要大量高纯度的毒素及其代谢产物。尽管这些毒素可从受污染的贝类或微藻培养物中获取少量,但当产毒微藻难以培养或其身份不明时,就会出现困难。为解决这一问题,我们开发了一种从海水中天然微藻水华中大规模固相萃取(SPE)亲脂性生物毒素的方法。为提高后续对吸附在柱上毒素的纯化效果,我们增加了一个过滤步骤,以便在去除不溶性化合物和细胞碎片的同时从细胞中释放毒素。从西班牙高密度的尖刺菱形藻水华以及挪威低密度尖刺菱形藻的混合水华中萃取和纯化冈田酸和鳍藻毒素-2,证明了该方法的有效性。吸附在SPE柱上的毒素的分离简单有效,目前获得的结果表明该方法可能适用于多种微藻毒素,如azaspiracids、pectenotoxins、spirolides和微囊藻毒素。该方法也应有助于从大规模微藻培养物中收获毒素,以及从海洋和淡水环境中进行生物活性天然产物的生物勘探和分离。