Engineering Research Center for Molecular Diagnosis, Faculty of Life Science and Technology, Kunming University of Science and Technology, Kunming 650500, Yunnan, China.
Anal Methods. 2024 Feb 29;16(9):1390-1398. doi: 10.1039/d3ay02065g.
Accidental ingestion of poisonous mushrooms leading to poisoning is a global issue. The most important and lethal toxin causing mushroom poisoning is α-amanitin, with a lethal dose of about 0.1 mg kg. Rapid detection of wild mushrooms before consumption or rapid identification of toxins after poisoning can effectively reduce the occurrence of fatalities. This study established a method for detecting α-amanitin using carbon dots/AuNPs nanoenzymes (D-Glu-CDs/AuNPs) with robust peroxidase-like activity. This nanoenzyme was prepared employing glucose carbon dots and sodium citrate as reducing and stabilizing agents, respectively. It could oxidize the substrate TMB (tetramethylbenzidine) to produce blue o-TMB. When α-amanitin specifically bound to the active site of the nanoenzyme, a resultant decrease was observed in catalytic activity and the absorbance value at 652 nm. The regression equation = -0.06083 + 0.9643, with an value of 0.996, was obtained. The limit of detection was determined to be 48.03 ng mL, and the recoveries in urine ranged from 91.2% to 97.6%. This method enabled the visualization of α-amanitin, and the whole detection process was completed within 20 min. The approach holds promise for the quantitative and qualitative determination of α-amanitin in urine samples.
误食毒蘑菇导致中毒是一个全球性问题。导致蘑菇中毒的最重要和最致命的毒素是α-鹅膏蕈碱，其致死剂量约为 0.1 毫克/千克。在食用前快速检测野生蘑菇或在中毒后快速鉴定毒素可以有效地降低死亡率的发生。本研究建立了一种使用具有强过氧化物酶样活性的碳点/AuNPs 纳米酶（D-Glu-CDs/AuNPs）检测α-鹅膏蕈碱的方法。该纳米酶采用葡萄糖碳点和柠檬酸钠分别作为还原剂和稳定剂制备。它可以将底物 TMB（四甲基联苯胺）氧化生成蓝色 o-TMB。当α-鹅膏蕈碱特异性结合到纳米酶的活性位点时，观察到催化活性和 652nm 处的吸光度值降低。得到回归方程为 = -0.06083 + 0.9643， 值为 0.996。检测限确定为 48.03ng/mL，尿液中的回收率在 91.2%至 97.6%之间。该方法实现了α-鹅膏蕈碱的可视化，整个检测过程在 20 分钟内完成。该方法有望用于尿液样本中α-鹅膏蕈碱的定量和定性测定。
