Beijing Advanced Innovation Center for Food Nutrition and Human Health, College of Veterinary Medicine, China Agricultural University, Beijing Key Laboratory of Detection Technology for Animal-Derived Food Safety, Beijing Laboratory for Food Quality and Safety, Beijing 100193, People's Republic of China.
Institute of Apicultural Research, Chinese Academy of Agricultural Sciences, Bee Product Quality Supervision and Testing Center, Laboratory of Risk Assessment for Quality and Safety of Bee Products, Ministry of Agriculture, Beijing 100093, People's Republic of China.
Anal Chim Acta. 2017 Nov 1;992:119-127. doi: 10.1016/j.aca.2017.09.010. Epub 2017 Sep 13.
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, 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 (R) 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.
本文描述了一种灵敏、快速且易于操作的荧光偏振免疫分析（FPIA）方法的建立，用于检测水中的微囊藻毒素（MCs）和节球藻毒素-R（NOD）。MCs 和 NOD 是最广泛存在的蓝藻毒素，属于肝毒素和肿瘤促进剂，其急性暴露可能会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，并首次采用制备高效液相色谱（HPLC）对其进行纯化。通过评估免疫化学活性，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）分别达到 0.86 和 0.95μg/L，灵敏度足以满足世界卫生组织（WHO）推荐的暂定指导值。建立了超高效液相色谱-串联质谱（UPLC-MS/MS）方法，以确认 FPIA 的准确性和精密度，两种方法的回收率无明显差异。相关性系数（R）均高于 0.968。开发的 FPIA 易于操作，水样经简单过滤和 pH 调整后，10 分钟内即可完成分析。该方法易于扩展用于筛选其他的蓝藻毒素，代表了一种用于环境样品分析的通用策略。
