Faculty of Chemistry, Razi University, Kermanshah, Iran; Biosensor Research Center, Endocrinology and Metabolism Molecular-Cellular Sciences Institute, Tehran University of Medical Sciences, Tehran, Iran.
Faculty of Chemistry, Sensor and Biosensor Research Center (SBRC) & Nanoscience and Nanotechnology Research Center (NNRC), Razi University, Kermanshah, Iran; Nanobiotechnology Department, Faculty of Innovative Science and Technology, Razi University, Kermanshah, Iran.
Spectrochim Acta A Mol Biomol Spectrosc. 2024 May 15;313:124057. doi: 10.1016/j.saa.2024.124057. Epub 2024 Feb 22.
A simple, highly sensitive, and selective fluorometric aptasensing platform based on aptamer and graphene oxide (GO) is proposed for the determination of mercury (II) ion (Hg). In the designed assay, two aptamer probes, a carboxy-fluorescein (FAM) labeled aptamer (aptamer A) and its complementary (aptamer B) with partial complement containing several mismatches and GO as the quencher were used. In the absence of Hg, both A and B aptamers were adsorbed on the surface of GO by π-π-stacking, leading to fluorescence quenching of FAM due to fluorescence resonance energy transfer (FRET). Upon exposure to Hg, the A and B aptamer strands bind Hg and form T-Hg-T complexes, leading to the formation of a stable double-stranded aptamer. The double-stranded aptamer is detached from the GO surface, resulting in the recovery of FAM fluorescence. The fluorescence intensity (FI) of the developed sensor was correlated with the Hg concentration under optimized experimental conditions in two wide linear ranges, even in the presence of 10 divalent cations as interferences. The linear ranges were obtained from 200.0 to 900.0 fM and 5.0 to 33.0 pM, a limit of detection (LOD) of 106.0 fM, and a limit of quantification (LOQ) of 321.3 fM. The concentration of Hg was determined in five real samples containing three water and two serum samples, using spiking and standard addition methods and the results were compared with the spiked amounts and atomic absorption (AAS) as standard method respectively, with acceptable recoveries. Furthermore, in the standard addition method, to overcome the effects of matrix influence of real samples in quantitative predictions, the excitation-emission matrix (EEM) data for samples was simultaneously analyzed by multivariate curve resolution with alternating least squares (MCR-ALS) as a second-order standard addition method (SOSAM).
基于适配体和氧化石墨烯(GO)的简单、高灵敏、选择性荧光适体传感平台被提出用于测定汞(II)离子(Hg)。在设计的测定中,使用两种适配体探针,一个羧基-荧光素(FAM)标记的适配体(适配体 A)及其互补链(带有几个错配的部分互补链,即适配体 B)和 GO 作为猝灭剂。在不存在 Hg 的情况下,A 和 B 两种适配体通过π-π 堆积吸附在 GO 表面上,由于荧光共振能量转移(FRET)导致 FAM 的荧光猝灭。当暴露于 Hg 时,A 和 B 适配体链与 Hg 结合形成 T-Hg-T 复合物,导致形成稳定的双链适配体。双链适配体从 GO 表面脱离,导致 FAM 荧光恢复。在优化实验条件下,该传感器的荧光强度(FI)与 Hg 浓度呈线性相关,在两个宽线性范围内,甚至在存在 10 种二价阳离子干扰的情况下也呈线性相关。线性范围从 200.0 至 900.0 fM 和 5.0 至 33.0 pM,检测限(LOD)为 106.0 fM,定量限(LOQ)为 321.3 fM。使用加标和标准加入法在五个含有三个水样和两个血清样的实际样品中测定了 Hg 的浓度,并分别与加标量和原子吸收(AAS)作为标准方法进行了比较,回收率是可接受的。此外,在标准加入法中,为了克服实际样品基质影响对定量预测的影响,通过交替最小二乘法(MCR-ALS)作为二阶标准加入法(SOSAM),同时对样品的激发-发射矩阵(EEM)数据进行多元曲线分辨分析。