Department of Biochemistry, College of Medicine, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan.
Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung, 80708, Taiwan.
Anal Bioanal Chem. 2021 Dec;413(30):7531-7539. doi: 10.1007/s00216-021-03717-6. Epub 2021 Oct 11.
Acrylamide is a group 2A carcinogen and potential endocrine disruptor that can enter the ecosystem by various routes and has recently become a dangerous pollutant. This widely used chemical can enter the human body via air inhalation, food or water consumption, or skin contact. In this study, we developed a peptide probe for the detection of acrylamide by matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) after its micro-tagging with a peptide. Direct detection of acrylamide by MALDI-TOF MS is not feasible due to its poor ionization in the MALDI interface, which hinders its analysis by the technique. After microwave irradiation for 2 min, the formed acrylamide-peptide derivative was detected easily by MALDI-TOF MS without the need for extraction procedures. The procedure does not involve organic solvents and a water-soluble peptide that allows detection of acrylamide in small sample volumes with a limit of detection (LOD) of 0.05 ng/μL. The relative standard deviation (RSD) and relative error (RE) of the measurements were < 6.7% for intra- and inter-day assays. Gel-washing solutions from a polyacrylamide gel experiment were used as a model to study the efficiency of the developed method. Finally, we used the proposed method for the detection of free acrylamide in small volumes of lung epithelial cells (a model to test the air inhalation of acrylamide under a tiny volume of sample) and human urine. The developed method will enable rapid acrylamide detection in environmental and biological samples via a green approach based on microwave-assisted derivatization in water alongside the use of a less toxic derivatization reagent, reusable target plate, and miniaturization protocols.
丙烯酰胺是一种 2A 类致癌物质和潜在的内分泌干扰物,它可以通过多种途径进入生态系统,最近已成为一种危险的污染物。这种广泛使用的化学物质可以通过空气吸入、食物或水摄入或皮肤接触进入人体。在这项研究中,我们开发了一种肽探针,用于通过基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)检测丙烯酰胺,方法是先对其进行肽微标记。由于在 MALDI 界面中其离子化效果不佳,直接通过 MALDI-TOF MS 检测丙烯酰胺是不可行的,这阻碍了该技术对其的分析。经微波辐射 2 分钟后,形成的丙烯酰胺-肽衍生物可通过 MALDI-TOF MS 轻松检测,无需提取程序。该程序不涉及有机溶剂和水溶性肽,允许在小样本量下检测丙烯酰胺,检出限(LOD)为 0.05ng/μL。日内和日间测定的相对标准偏差(RSD)和相对误差(RE)均<6.7%。将聚丙烯酰胺凝胶实验的凝胶洗涤溶液用作模型,以研究所开发方法的效率。最后,我们使用所提出的方法检测小体积肺上皮细胞(模型以测试在小体积样本下空气中吸入丙烯酰胺的情况)和人尿中的游离丙烯酰胺。该方法将通过基于微波辅助水相衍生化的绿色方法以及使用毒性较小的衍生化试剂、可重复使用的靶板和小型化方案,在环境和生物样品中实现快速丙烯酰胺检测。