A colorimetric and fluorometric microplate assay for the detection of microcystin-LR in drinking water without preconcentration
A colorimetric and fluorometric microplate assay for the detection of microcystin-LR in drinking water without preconcentration
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DOI：
10.1016/s0278-6915(02)00103-5
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发表时间：
2002-11-01
影响因子：
4.3
通讯作者：
Levi, Y
中科院分区：
文献类型：
--
作者：
Bouaïcha, N;Maatouk, I;Levi, Y
目前用于检测饮用水中蓝藻多肽肝毒素的蛋白磷酸酶抑制法需要使用C18小柱进行浓缩步骤，以实现较低的检测限。本文介绍了一种直接测定饮用水中微囊藻毒素-LR(MCYST-LR)的比色荧光蛋白磷酸酶抑制法，不需要复杂的净化步骤和预浓缩步骤。在本实验中，测试了三种不同的底物对硝基苯基磷酸(p-NPP)和两种含氟化合物4-甲基伞形基磷酸酯(MUP)和6，8-二氟-4-甲基伞形基磷酸酯(DiFMUP)。比色法和荧光法的检出限分别为0.25ug/L和0.1ug/100g。这些水平远低于1杯/L饮用水的MCYST-LR暂定指导值。该荧光法的检出限与经典的酶联免疫吸附试验相当。虽然后两种检测方法都可以在不经过前处理的情况下直接检测饮用水中的MCYST-LR，但蛋白磷酸酶抑制试验的成本较低，因此更适合用于饮用水中微囊藻毒素污染的常规评估。(C)2002爱思唯尔科学有限公司。保留所有权利。
Protein phosphatase inhibition assays currently used for the detection of cyanobacterial peptide hepatotoxins in drinking water require an enrichment step using C18 cartridges to achieve lower the detection limit. This paper describes a colorimetric and fluorometric protein phosphatase inhibition method for the direct detection of microcystin-LR (MCYST-LR) in drinking water without complex clean-up steps and preconcentration procedures. In this assay three different substrates, p-nitrophenyl phosphate (p-NPP) and two fluorogenic compounds, 4-methylumbelliferyl phosphate (MUP) and 6,8-difluoro-4-methylumbelliferyl phosphate DiFMUP), were tested. The detection limits of the assay are 0.25 and 0.1 mug/l using colorimetric and fluorometric methods, respectively. These levels are well below the provisional guideline value for MCYST-LR of 1 mug/l of drinking water. The detection limit of the fluorometric method is comparable to that of the classical ELISA test. Although both the latter tests allow the detection of MCYST-LR in drinking water directly without pretreatment, the protein phosphatase inhibition assay remain less expensive and therefore more attractive for use in the routine assessment of drinking water contamination by microcystins. (C) 2002 Elsevier Science Ltd. All rights reserved.