This manuscript describes the development of a sensitive, fast and easily-performed fluorescence polarization immunoassay (FPIA) for detection of microcystins (MCs) and nodularin-R (NOD) in water. MCs and NOD, the most widespread cyanobacterial toxin are hepatotoxins and tumor promoters, and their acute exposure may result in severe health problems in animals and humans. The fluorescein-based tracers were synthesized, and for the first time preparative high performance liquid chromatography (HPLC) was employed for their purification. Optimal tracers for the analysis were selected by evaluating the immunochemical activity. Under the optimal conditions, the achieved limits of detection (LODs) for MC-LR and NOD were 0.86 and 0.95 μg L-1, respectively, providing a sufficient sensitivity to meet the provisional guideline value recommended by the World Health Organization (WHO). An ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to confirm the accuracy and precision of the FPIA, and no obvious difference in recovery between these two methods was found. The correlation coefficients (R2) were higher than 0.968. The developed FPIA was easy-to-operate and could be completed within 10 min after simple filtration and adjustment of pH for water samples. The method can be easily extended for screening of other cyanotoxins, representing a versatile strategy for environmental sample analysis.

中文翻译：

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通过荧光偏振免疫分析简单、高效地检测水中的微囊藻毒素和节球藻毒素-R

本手稿描述了一种灵敏、快速且易于执行的荧光偏振免疫分析 (FPIA) 的开发，用于检测水中的微囊藻毒素 (MC) 和节球藻毒素-R (NOD)。MCs 和 NOD，最普遍的蓝藻毒素是肝毒素和肿瘤促进剂，它们的急性暴露可能导致动物和人类的严重健康问题。合成了基于荧光素的示踪剂，并首次采用制备型高效液相色谱 (HPLC) 对其进行纯化。通过评估免疫化学活性来选择用于分析的最佳示踪剂。在最佳条件下，MC-LR 和 NOD 的检测限 (LOD) 分别为 0.86 和 0.95 μg L-1，提供足够的灵敏度以满足世界卫生组织 (WHO) 推荐的临时指南值。开发了一种超高效液相色谱-串联质谱 (UPLC-MS/MS) 方法来确认 FPIA 的准确度和精密度，发现这两种方法之间的回收率没有明显差异。相关系数 (R2) 高于 0.968。所开发的FPIA操作简单，只需简单过滤和调整水样pH值即可在10分钟内完成。该方法可以很容易地扩展到筛选其他蓝藻毒素，代表了环境样品分析的通用策略。开发了一种超高效液相色谱-串联质谱 (UPLC-MS/MS) 方法来确认 FPIA 的准确度和精密度，发现这两种方法之间的回收率没有明显差异。相关系数 (R2) 高于 0.968。所开发的FPIA操作简单，只需简单过滤和调整水样pH值即可在10分钟内完成。该方法可以很容易地扩展到筛选其他蓝藻毒素，代表了环境样品分析的通用策略。开发了一种超高效液相色谱-串联质谱 (UPLC-MS/MS) 方法来确认 FPIA 的准确度和精密度，发现这两种方法之间的回收率没有明显差异。相关系数 (R2) 高于 0.968。所开发的FPIA操作简单，只需简单过滤和调整水样pH值即可在10分钟内完成。该方法可以很容易地扩展到筛选其他蓝藻毒素，代表了环境样品分析的通用策略。