Effect of pretreatment approach on the ELISA-based detection of cyanotoxins in water: Analysis and application

Abstract

Common cyanotoxins, such as microcystins and nodularins, are produced by frequently occurring harmful cyanobacterial algal blooms in freshwater systems. The required routine monitoring of microcystins and nodularins in drinking water and ambient water demands cost-efficient and reliable enzyme-linked immunosorbent assay kits. We validated the performance of a self-produced broad-spectrum enzyme-linked immunosorbent assay kit and investigated two different methods of mitigating the matrix effects to elucidate the effect of the respective pretreatment approaches recommended by China and the United States on the quantitative detection of cyanotoxins in surface water. We found that the enzyme-linked immunosorbent assay kit achieved a detection limit of 0.15 μg/L with a linear detection range from 0.27 μg/L to 1.87 μg/L for microcystin-LR (the most studied and widely distributed cyanotoxin). The matrix effects could be mitigated both by dilution of water samples with an optimal dilution ratio and dilution of antibody with the buffer containing phosphate buffer solution (10×), bovine serum albumin (1 %) and ethylene diamine tetraacetic acid (0.5 %). In terms of the surface water samples being tested, the concentrations of microcystins and nodularins measured based on pretreatment approach recommended by the United States were 1– 5 times that measured based on pretreatment approach recommended by China, indicating that the pretreatment approach of China overlooks cyanotoxins. In addition, all the measured total microcystins and nodularins of the surface water samples were below the health advisory limit (1.6 μg/L) for microcystins in drinking water proposed by the United States Environmental Protection Agency for school-age children and adults. Our research could provide significant information for outbreak warnings and risk management of harmful cyanobacterial algal blooms.