Emergency Response Branch, Division of Laboratory Sciences, National Center for Environmental Health, Centers for Disease Control and Prevention, Atlanta, GA 30341, USA.
Battelle Memorial Institute, Centers for Disease Control and Prevention, 1600 Clifton Road, Atlanta, GA 30333, USA.
Toxins (Basel). 2024 Mar 1;16(3):129. doi: 10.3390/toxins16030129.
Harmful cyanobacterial blooms are becoming more common and persistent around the world. When in bloom, various cyanobacterial strains can produce anatoxins in high concentrations, which, unlike other cyanobacterial toxins, may be present in clear water. Potential human and animal exposures to anatoxins occur mainly through unintentional ingestion of contaminated algal mats and water. To address this public health threat, we developed and validated an LC-MS/MS method to detect anatoxins in human urine to confirm exposures. Pooled urine was fortified with anatoxin-a and dihydroanatoxin at concentrations from 10.0 to 500 ng/mL to create calibrators and quality control samples. Samples were diluted with isotopically labeled anatoxin and solvent prior to LC-MS/MS analysis. This method can accurately quantitate anatoxin-a with inter- and intraday accuracies ranging from 98.5 to 103% and relative standard deviations < 15%, which is within analytical guidelines for mass spectrometry methods. Additionally, this method qualitatively detects a common degradation product of anatoxin, dihydroanatoxin, above 10 ng/mL. We also evaluated a commercial anatoxin-a ELISA kit for potential diagnostic use; however, numerous false positives were detected from unexposed individual human urine samples. In conclusion, we have developed a method to detect anatoxins precisely and accurately in urine samples, addressing a public health area of concern, which can be applied to future exposure events.
有害的蓝藻水华在世界各地变得越来越普遍和持久。当处于水华期时，各种蓝藻菌株可以高浓度地产生anatoxins，与其他蓝藻毒素不同，anatoxins 可能存在于清澈的水中。人类和动物潜在地暴露于 anatoxins 主要是通过意外摄入受污染的藻类垫和水。为了解决这一公共卫生威胁，我们开发并验证了一种 LC-MS/MS 方法，以检测人尿中的 anatoxins 来确认暴露情况。将 anatoxin-a 和二氢anatoxin 与尿液混合，浓度从 10.0 到 500ng/ml，以创建校准品和质控样品。样品在 LC-MS/MS 分析前用同位素标记的 anatoxin 和溶剂稀释。该方法可以准确地定量 anatoxin-a，日内和日间准确度范围为 98.5%至 103%，相对标准偏差<15%，符合质谱方法的分析指南。此外，该方法还可以定性地检测到 anatoxin 的一种常见降解产物，二氢anatoxin，浓度高于 10ng/ml。我们还评估了一种商业的 anatoxin-a ELISA 试剂盒，用于潜在的诊断用途；然而，从未暴露的个体人尿样中检测到许多假阳性。总之，我们已经开发出一种在尿液样本中精确和准确地检测 anatoxins 的方法，解决了一个令人关注的公共卫生领域的问题，该方法可应用于未来的暴露事件。
