Multi-Laboratory Study of Five Methods for the Determination of Brevetoxins in Shellfish Tissue Extracts.

Harmful algae 2002 : proceedings of the Xth International Conference on Harmful Algae, St. Pete Beach, Florida, USA, October 21-25, 2002. International Conference on Harmful Algae (10th : 2002 : St. Pete Beach, Florida) Pub Date : 2004-01-01

Robert W Dickey, Steven M Plakas, Edward L E Jester, Kathleen R El Said, Jan N Johannessen, Leanne J Flewelling, Paula Scott, Dan G Hammond, Frances M Van Dolah, Tod A Leighfield, Marie-Yasmine Bottein Dachraoui, John S Ramsdell, Richard H Pierce, Mike S Henry, Mark A Poli, Calvin Walker, Jan Kurtz, Jerome Naar, Daniel G Baden, Steve M Musser, Kevin D White, Penelope Truman, Aaron Miller, Timothy P Hawryluk, Marleen M Wekell, David Stirling, Michael A Quilliam, Jung K Lee

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Abstract

A thirteen-laboratory comparative study tested the performance of four methods as alternatives to mouse bioassay for the determination of brevetoxins in shellfish. The methods were N2a neuroblastoma cell assay, two variations of the sodium channel receptor binding assay, competitive ELISA, and LC/MS. Three to five laboratories independently performed each method using centrally prepared spiked and naturally incurred test samples. Competitive ELISA and receptor binding (96-well format) compared most favorably with mouse bioassay. Between-laboratory relative standard deviations (RSDR) ranged from 10 to 20% for ELISA and 14 to 31% for receptor binding. Within-laboratory (RSDr) ranged from 6 to 15% for ELISA, and 5 to 31% for receptor binding. Cell assay was extremely sensitive but data variation rendered it unsuitable for statistical treatment. LC/MS performed as well as ELISA on spiked test samples but was inordinately affected by lack of toxin-metabolite standards, uniform instrumental parameters, or both, on incurred test samples. The ELISA and receptor binding assay are good alternatives to mouse bioassay for the determination of brevetoxins in shellfish.

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贝类组织提取物中短链毒素五种测定方法的多实验室研究。

一项由13个实验室组成的比较研究测试了四种方法作为小鼠生物测定法测定贝类中短毒素的替代方法的性能。方法为N2a神经母细胞瘤细胞法、两种钠通道受体结合法、竞争性ELISA法和LC/MS法。三到五个实验室使用中央制备的加标剂和自然产生的测试样品独立执行每种方法。竞争性ELISA和受体结合(96孔格式)与小鼠生物测定法相比最有利。ELISA的实验室间相对标准偏差(RSDR)为10% - 20%，受体结合的实验室间相对标准偏差为14% - 31%。ELISA的实验室内(RSDr)为6 - 15%，受体结合的实验室内(RSDr)为5 - 31%。细胞分析是非常敏感的，但数据的变化使其不适合统计处理。LC/MS在加标测试样品上的表现与ELISA一样好，但在产生的测试样品上，由于缺乏毒素代谢物标准，统一的仪器参数或两者兼而有之，因此受到极大的影响。ELISA和受体结合法是替代小鼠生物测定法测定贝类中短毒素的较好方法。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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Harmful algae 2002 : proceedings of the Xth International Conference on Harmful Algae, St. Pete Beach, Florida, USA, October 21-25, 2002. International Conference on Harmful Algae (10th : 2002 : St. Pete Beach, Florida)

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