Hesp Blair R, Harrison Joanne C, Selwood Andrew I, Holland Patrick T, Kerr D Steven
Department of Pharmacology & Toxicology, University of Otago, School of Medical Sciences, P.O. Box 913, Dunedin, New Zealand.
Anal Bioanal Chem. 2005 Nov;383(5):783-6. doi: 10.1007/s00216-005-0060-3. Epub 2005 Nov 5.
In 1987 a large-scale incident of human poisoning in Canada was traced to commercial mussels contaminated with domoic acid (DOM). Since then, routine screening of shellfish domoic acid content has been carried out using a variety of assays, with liquid chromatography using ultraviolet absorbance detection (LC-UV) or mass spectrometric detection (LC-MS) being the currently accepted standard methodologies. Recently, a highly specific competitive enzyme-linked immunosorbent assay (cELISA) has been developed for the detection and analysis of DOM in commercial shellfish, but its accuracy relative to LC methods has not been independently verified in mammalian tissues. In this study we demonstrate that measurement of rat serum DOM concentration by cELISA gives a good correlation (r2 = 0.993) across a broad range of concentrations when compared to LC-MS analysis, with only a small (15%) overestimation of sample DOM content. In addition, we have developed an extraction method for analysis of DOM in rat brain by cELISA which yields complete recovery across a range of sample dilutions.
1987年，加拿大发生了一起大规模的人类中毒事件，经追溯是由受软骨藻酸（DOM）污染的商业贻贝所致。从那时起，人们使用各种检测方法对贝类中的软骨藻酸含量进行常规筛查，目前公认的标准方法是采用紫外吸收检测的液相色谱法（LC-UV）或质谱检测的液相色谱法（LC-MS）。最近，一种用于检测和分析商业贝类中DOM的高特异性竞争酶联免疫吸附测定法（cELISA）已被开发出来，但在哺乳动物组织中其相对于LC方法的准确性尚未得到独立验证。在本研究中，我们证明，与LC-MS分析相比，采用cELISA测定大鼠血清DOM浓度时，在很宽的浓度范围内都具有良好的相关性（r2 = 0.993），对样品DOM含量的高估仅为15%。此外，我们还开发了一种通过cELISA分析大鼠脑中DOM的提取方法，该方法在一系列样品稀释度下均能实现完全回收。
