Detection of microcystins with protein phosphatase inhibition assay, high-performance liquid chromatography-UV detection and enzyme-linked immunosorbent assay - Comparison of methods
Detection of microcystins with protein phosphatase inhibition assay, high-performance liquid chromatography-UV detection and enzyme-linked immunosorbent assay - Comparison of methods
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DOI：
10.1016/s0003-2670(02)00588-3
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发表时间：
2002-08-27
影响因子：
6.2
通讯作者：
Lahti, K
中科院分区：
文献类型：
--
作者：
Rapala, J;Erkomaa, K;Lahti, K
比较了蛋白磷酸酶抑制比色法（PPI法）、酶联免疫吸附试验（ELISA法）和不同的高效液相色谱法（HPLC法）对蓝藻肝毒素、微囊藻毒素（MCYST）和节球藻毒素的检测效果。通过使用纯毒素和实验室培养物以及有毒蓝藻的水和水华样品来评估检测不同毒素变体的方法的适用性。这项研究的重点是分析自然界中常见但没有商业标准的极性去甲基微囊藻毒素变体。MCYST-LR对PPI测定和ELISA试验的IC 50值分别为2.2-2.5和0.26-0.38 μ g-1。减少样品处理中毒素回收率的最重要因素是使用C-18药筒和聚丙烯容器。通过使用亲水亲油平衡（Oasis HLB，沃茨）柱浓缩样品，获得了良好的毒素回收率。除[D-Asp（3）]微囊藻毒素外，PPI测定、ELISA测定和HPLC测定结果在定量上具有良好的相关性。PPI测定法测得的[D-ASP（3）]MCYST-RR浓度仅为ELISA试验和HPLC法测得浓度的5%。与其他方法相比，用ELISA分析时，疏水性微囊藻毒素变体的浓度较低。世界卫生组织（WHO）已经为饮用水中世界范围内最常见的微囊藻毒素变体MCYST-LR设定了1 mug 1（-1）的指导值。由于PPI检测和ELISA检测的定量范围在天然沃茨中的微囊藻毒素浓度范围内，并且这两种检测都很容易进行，因此它们显示出在饮用水供应和娱乐沃茨中微囊藻毒素的筛选和监测中常规使用的潜力。(C)出版社：Elsevier Science B. V.
A colorimetric protein phosphatase inhibition assay (PPI assay), a commercial enzyme-linked immunosorbent assay (ELISA) test and different HPLC methods using UV detection were compared for the detection of cyanobacterial hepatotoxins, microcystins (MCYST) and nodularin. The suitability of the methods to detect different toxin variants was evaluated by using pure toxins and laboratory cultures as well as water and bloom samples of toxic cyanobacteria. The emphasis of the study was on the analysis of polar demethyl microcystin variants that are common in nature but for which there exist no commercial standards. The IC50 values of MCYST-LR for the PPI assay and the ELISA test were 2.2-2.5 and 0.26-0.38 mug 1(-1), respectively. The most important factors that decreased toxin recovery in sample treatment were the use of C-18 cartridges and polypropylene containers. Good recoveries of toxins were obtained by using hydrophilic-lipophilic balanced (Oasis HLB, Waters) cartridges for concentrating the samples. The results obtained with the PPI assay, the ELISA test and HPLC correlated quantitatively well with the exception Of [D-Asp(3)] microcystins. Concentrations Of [D-ASp(3)]MCYST-RR measured with the PPI assay were only 5% of those obtained by the ELISA test and HPLC. Concentrations of hydrophobic microcystin variants were lower when analysed with ELISA than with the other methods. The World Health Organisation (WHO) has set a guideline value of 1 mug 1(-1) for the world-wide most common microcystin variant, MCYST-LR in drinking water. Since the quantitative ranges of the PPI assay and the ELISA test are within microcystin concentrations in natural waters, and both tests are easy to perform, they show potential for routine use in the screening and monitoring of microcystins from drinking water supplies and from recreational waters. (C) 2002 Published by Elsevier Science B.V.