Indrasena W M, Gill T A
Department of Food Science and Technology, DalTech, Dalhousie University, Halifax, Nova Scotia, Canada.
Anal Biochem. 1998 Nov 15;264(2):230-6. doi: 10.1006/abio.1998.2843.
A rapid qualitative screening method was developed for the fractionation of paralytic shellfish poisoning toxins. Periodic acid, t-butyl hydroperoxide, and hydrogen peroxide were tested as oxidants for the fluorometric detection of paralytic shellfish toxins. Hydrogen peroxide was found to be the most convenient and efficient oxidant since the fluorescence can be detected after the incubation of toxins at 100 degrees C for 3-5 min. In addition to the structure of the compound, the incubation temperature and time, the amount of acid, and the peroxide concentration affect the fluorescence reaction. This method was more efficient than the previously published peroxidation methods which involved lengthy incubation periods or time-consuming pH adjustment. Also, far greater sensitivity was achieved with the new method with levels of 0.027, 0.054, 0.023, 0.003, 0.0002, and 0.0006 pmol being easily detected for saxitoxin, neosaxitoxin, gonyautoxin 1 and 4, gonyautoxin 2 and 3, C toxins, and B toxins, respectively. The method is particularly valuable for the screening of fractions separated by column chromatography.
开发了一种用于麻痹性贝类中毒毒素分级分离的快速定性筛选方法。高碘酸、叔丁基过氧化氢和过氧化氢作为氧化剂用于麻痹性贝类毒素的荧光检测进行了测试。发现过氧化氢是最方便且高效的氧化剂,因为毒素在100℃孵育3 - 5分钟后即可检测到荧光。除化合物结构外,孵育温度和时间、酸的用量以及过氧化物浓度都会影响荧光反应。该方法比先前发表的涉及长时间孵育或耗时pH调节的过氧化方法更有效。此外,新方法具有更高的灵敏度,分别可轻松检测到0.027、0.054、0.023、0.003、0.0002和0.0006皮摩尔的石房蛤毒素、新石房蛤毒素、膝沟藻毒素1和4、膝沟藻毒素2和3、C毒素和B毒素。该方法对于柱色谱分离组分的筛选特别有价值。