State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, Jilin, P. R. China.
University of Science and Technology of China, Hefei 230026, Anhui, P. R. China.
Anal Chem. 2022 Mar 8;94(9):4112-4118. doi: 10.1021/acs.analchem.2c00124. Epub 2022 Feb 23.
Some mustard compounds (mustards) are highly toxic chemical warfare agents. Some are explored as new anticancer drugs. Therefore, the fast, selective, and sensitive detection of mustards is extremely important for public security and cancer therapy. Mustards mostly target the N7 position on the guanine bases of DNA. The guanine-rich G-quadruplex DNA (G4) has been widely studied in the sensing area, and it was found that dimeric G4 (D-G4) could dramatically light up the fluorescence intensity of thioflavin T (ThT). Based on this, we used for the first time the D-G4 DNA as a selective probe for ultrasensitive fluorescence detection of nitrogen mustard (NM). When NM occupies the N7 on guanine, it can block the formation of the D-G4 structure due to the steric hindrance, and hence, it inhibits the combination of D-G4 with ThT, leading to a sharp decrease of fluorescence intensity. The proposed reaction mechanism is proved using ultraviolet-visible (UV-Vis) spectra, circular dichroism (CD) spectra, and polyacrylamide gel electrophoresis. Herein, the concentration of D-G4/ThT used is as low as 50 nM due to its highly fluorescent performance, enabling both high sensitivity and low cost. NM can be detected with a wide linear range from 10 to 2000 nM. The detection limit of NM reaches a surprisingly low concentration of 6 nM, which is 2 or 3 orders of magnitude lower than that of previously developed fluorescence methods for mustards and simulants.
一些芥子气化合物(芥子气)是剧毒的化学战剂。有些被探索作为新型抗癌药物。因此,快速、选择性和灵敏地检测芥子气对于公共安全和癌症治疗至关重要。芥子气主要靶向 DNA 中鸟嘌呤碱基的 N7 位置。富含鸟嘌呤的 G-四链体 DNA(G4)在传感领域得到了广泛研究,发现二聚 G4(D-G4)可以显著增强硫黄素 T(ThT)的荧光强度。基于此,我们首次将 D-G4 DNA 用作选择性探针,用于超灵敏荧光检测氮芥(NM)。当 NM 占据鸟嘌呤上的 N7 时,由于空间位阻,它可以阻止 D-G4 结构的形成,从而抑制 D-G4 与 ThT 的结合,导致荧光强度急剧下降。使用紫外-可见(UV-Vis)光谱、圆二色性(CD)光谱和聚丙烯酰胺凝胶电泳证明了该反应机制。由于其高荧光性能,这里使用的 D-G4/ThT 浓度低至 50 nM,实现了高灵敏度和低成本。NM 可以在 10 至 2000 nM 的宽线性范围内检测到。NM 的检测限达到了惊人的低浓度 6 nM,比以前开发的芥子气和模拟物的荧光方法低 2 到 3 个数量级。