Detection of domoic acid in rat serum and brain by direct competitive enzyme-linked immunosorbent assay (cELISA).
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.
Hesp
Blair R
BR
Department of Pharmacology & Toxicology, University of Otago, School of Medical Sciences, P.O. Box 913, Dunedin, New Zealand.
Harrison
Joanne C
JC
Selwood
Andrew I
AI
Holland
Patrick T
PT
Kerr
D Steven
DS
D000818:Animals; D001921:Brain / Q000378:metabolism*; D002853:Chromatography, Liquid / Q000379:methods*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D007608:Kainic Acid / Q000031:analogs & derivatives* / Q000032:analysis / Q000097:blood / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D008832:Microchemistry / Q000379:methods*; D051381:Rats; D017207:Rats, Sprague-Dawley
C012301:domoic acid; D007608:Kainic Acid